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  • Table of Content
      28 March 2019, Volume 35 Issue 2 Previous Issue    Next Issue
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    ORIGINAL ARTICLES
    Effects of acute ozone exposure on genotoxicity of lung cells in rats
    LI Ning, YANG Hu, FANG Zhen, WANG Ping-yu, HAN Jie, TIAN Lei, YAN Jun, XI Zhu-ge, LIU Xiao-hua
    CJAP. 2019, 35 (2): 97-100.   DOI: 10.12047/j.cjap.5738.2019.022
    Abstract   PDF (1162KB) ( 160 )
    Objective: To clarify the genotoxicity induced by acute exposure of ozone with different concentrations on pulmonary cells in rats. Methods: Thirty-six Wistar rats were randomly divided into control group (filtered air exposure) and ozone exposure group (0.12 ppm, 0.5 ppm, 1.0 ppm, 2.0 ppm, 4.0 ppm) with 6 in each group. After rats were exposed to different concentrations of ozone for 4 h, lung tissues were taken and single cells were isolated. Then, 8-hydroxydeoxyguanosine (8-OHdG) was quantitatively detected by enzyme-linked immunosorbent assay. Comet assay, micronucleus test and DNA- protein cross-linking assay were used to analyze DNA and chromosome damages. Results: Compared with the control group, the content of 8-OHdG in lung tissue was increased significantly from the ozone exposure concentration of 0.12 ppm, reaching the highest value at 0.5 ppm. With the increase of ozone exposure concentration, the tail rate of comets was increased gradually, and there was a significant dose-effect relationship. The cross-linking rate of DNA- protein was increased first and then was decreased with a maximum value at 2.0 ppm group. Although the micronucleus rate of lung cells showed an upward trend, there was no significant difference compared with the control group. Conclusion: Acute exposure of ozone at low concentrations (0.12 ppm) could lead to DNA damage in the pulmonary cells of rats, while no significant chromosome damage was found even in the group with ozone concentration reached to 4 ppm.
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    Effects of Yiqi Huayu Hutan decoction on pulmonary fibrosis in rats and its mechanism
    TIAN Sheng, CAO Wen-fu, ZHANG Yong-yue, WU Qing
    CJAP. 2019, 35 (2): 101-106.   DOI: 10.12047/j.cjap.5711.2019.023
    Abstract   PDF (2285KB) ( 239 )
    Objective: To investigate the effects of Yiqi Huayu Hutan decoction on pulmonary fibrosis of rats which induced by bleomycin. Methods: The rat model of pulmonary fibrosis was induced by intratracheal injection of bleomycin hydrochloride (5 mg/kg). Sixty SD rats were randomly divided into the normal group (group N), the model group (group M), the positive control group (group Y), group of low concentration (group LC), group of medium concentration (group MC) and group of high concentration of Yiqi Huayu Hutan decoction (group HC). After 4 weeks, the experimental groups were treated with low concentration decoction, medium concentration decoction and high concentration decoction respectively, and the Y group was treated with hydrocortisone acetate, the Group N and group M were treated with saline by intragastric administration. Twelve weeks later, rats were killed and the pathomorphism of pulmonary tissues of each group was observed by HE staining and Masson staining. Further, the expressions of transforming growth factor-β1(TGF-β1), Snail1, E-cadherin and Fibronectin in pulmonary tissues of each group were detected by qTR-PCR and Western blot. Results: Compared with the model group, the collagen sediment in the interstitial was reduced in the experimental groups, especially in the group of medium concentration, which was observed by HE staining and Masson staining .Compared with the model group, the expressions of TGF-β1, Snail1 and Fibronectin protein in pulmonary tissues of the treatment groups were decreased in the experimental group, especially in the group of medium concentration, which were detected by qRT-PCR and Western blot. Conclusion: Yiqi Huayu Hutan decoction can significantly improve the pulmonary fibrosis which is induced by bleomycin, and the mechanism is related to the inhibition of the expression of TGF-β/Snail pathway of transcription TGF-β1.
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    The interventional effects of saturated hydrogen saline on lung injury in rats with cecal ligation and puncture operation
    ZHAI Yu, ZHOU Xiao-hong, LIU Hui, FU Hong-yi, FAN Ya-min, HUANG Xin-li
    CJAP. 2019, 35 (2): 107-112.   DOI: 10.12047/j.cjap.5767.2019.024
    Abstract   PDF (1250KB) ( 192 )
    Objective: To investigate the hypothesis that hydrogen could ameliorate cecal ligation and puncture (CLP)-induced lung injury of rats by inhibiting cystathionine-gamma-lyase/hydrogen sulfide (CSE/H2S) system. Methods: A total number of 24 healthy male SD rats weighting 250~300 g were randomly divided into four groups (n=6 in each group): sham operation group(sham group), hydrogen-rich saline control group(H2 group), CLP group and hydrogen-rich saline treatment group(CLP+H2 group). The rats were treated with hydrogen-rich saline or saline 10 min before CLP or sham operation. At 8 h of sham or CLP operation, lung samples were obtained to detect the changes of the CSE/H2S system using biochemical and RT-PCR methods. In order to further confirm the role of H2S during hydrogen improve the lung injury of CLP rats, we also observed the effect of hydrogen-rich saline on the lung injury induced by H2S donor-sodium sodium hydrosulfide (NaHS). Thirty-two healthy male SD rats (250~300 g) were randomly divided into four groups (n=8 in each group): control group, H2S group, H2S+H2 group and H2 group. Saline(10 mg/kg) or NaHS(H2S donor, 56 μmol/kg) was injected intraperitoneally (10 mg/kg) respectively into rats in the control rats or H2S group. For rats in the H2S+H2 and H2 group, hydrogen-rich saline (10 mg/kg) was injected 10 min before saline or NaHS administration. Eight hours after the LPS saline or NaHS administration, lung coefficient, MDA content, and MPO activity were detected. The contents of TNF-α, IL-6 and IL-10 in lung tissue were measured, and the morphological changes of lung tissue were also observed. Results: CSE/H2S system up-regulating were observed in animals exposed to CLP. Hydrogen-rich saline treatment significantly inhibited CSE/H2S system as indicated by significantly reduced H2S production in lung, along with a decreased CSE activity and CSE mRNA expression (all P<0.05). Importantly, the results showed that lung injury and lung tissue inflammation were observed in animals exposed to NaHS. Hydrogen-rich saline treatment significantly attenuated lung injury as indicated by significantly improved histological changes in lung, significantly reduced index of quantitative assessment (IQA), MDA content and lung coefficient (all P<0.05). MPO activity in lung tissue was significantly reduced along with decreased productions of TNF-α and IL-6, and an increased production of IL-10 in the presence of hydrogen (all P<0.05), demonstrating antioxidant and anti-inflammatory effect of hydrogen in NaHS-induced ALI. Conclusion: These results indicate that hydrogen-rich saline peritoneal injection improves the lung injury induced by CLP operation. The therapeutic effects of hydrogen-rich saline may be related to suppressing the production of H2S.
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    Effects of interval training on calcium transient and contractile function of single ventricular myocyte in myocardial infarction adult rats
    BO Wen-yan, LI Da-gang, TIAN Zhen-jun
    CJAP. 2019, 35 (2): 121-125.   DOI: 10.12047/j.cjap.5721.2019.027
    Abstract   PDF (1487KB) ( 216 )
    Objective: To investigate the effects of interval training on calcium transient and contractile function in ischemic ventricular myocytes of rats with myocardial infarction and their synchronization. Methods: Twenty-four male sprague-dawley rats in three years old, were randomly divided into three groups (n=8): sham-operated group(S), sedentary MI group(MI) and MI with interval training group (ME). The MI model was established by ligation of the left anterior descending coronary artery. The rats in ME group started training 1 week after MI operation. The S model was established by threading only without ligation. ME model took one week adaptive training, 10 m/min and 30 min/d, then took subsequently 8-week aerobic interval training: 10 min×10 m/min, then reran the rats with 2 intensities 15 m/min×6 min and 25 m/min×4 min, 1 h/d, 5 d/week. After training 24 hours, the cardiomyocytes of all groups were isolated by using the Langendorff fusion system. The contractile function and calcium transient of single ventricular myocyte in myocardial infarction adult rats were detected by IonOptix. Calcium transients were measured as [Ca2+]i amplitude, departure velocity, ratio, TTB50%, TTP and TTP50%, return velocity and ratio amplitude. PTA, SL,±dl/dtmax and SL shortening% were tested to evaluate contractility. Results: Compared with S, the levels of [Ca2+]i amplitude, departure velocity, ratio amplitude and return velocity, SL shortening%, PTA and±dl/dtmax of MI were decreased(P<0.01), the levels of TTP, TTP50% and TTB50% of MI were increased(P<0.01); Compared with MI, the levels of departure velocity, ratio amplitude, return velocity and [Ca2+]i amplitude of ME were increased(P<0.01), the levels of TTB50%, TTP and TTP50% of ME were decreased(P<0.01, P<0.05). The levels of SL shortening%, PTA and±dl/dtmax of ME were increased(P<0.01, P<0.05). Conclusion: Interval training can improve calcium transient and contractile function of single ventricular myocyte in myocardial infarction adult rats.
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    The effects of lower limb intermittent negative pressure therapy on the skin microcirculation perfusion of quadriceps in male rowers
    DONG Hui-hui, GAO Bing-hong, ZHU Huan, YANG Sheng-tao
    CJAP. 2019, 35 (2): 126-129.   DOI: 10.12047/j.cjap.5727.2019.028
    Abstract   PDF (920KB) ( 231 )
    Objective: To investigate the effects of intermittent negative pressure therapy on the skin microcirculation perfusion of quadriceps in male rowers, and to provide basis for the practical application of this method. Methods: Fourteen male rowers were selected from the national rowing team and randomly divided into experimental and control groups. The daily training plans of two groups were the same. The recovery intervention for experimental group was implemented by 20 minutes in the cube of Vacusport Regeneration System (German), 5 times per week for 4 weeks, no recovery intervention for control group. Microcirculation markers were collected by PeriFlux5000 system before and after the 4-week intervention. The markers included microcirculatory blood perfusion(MBP), average velocity of blood cells(AVBC), concentration of moving blood cells (CMBC), and values of the markers included basic values and post-heating values (44℃), difference before and after heating of the values was considered as the reserve capacity of those markers. Results: The test results before the 4 weeks intervention showed there was no statistical difference between the two groups(P>0.05). After the 4 weeks intervention: ①MBP: The post-heating value and the difference of the experimental group were significantly higher than those of the control group (P<0.05). But there was no statistical inner-group difference. ②AVBC: The post-heating values and the difference in the experimental group were significantly lower than those in the control group (P<0.05). Intra-group comparison found that the post-heating values after post-intervention were significantly reduced, compared with those of pre-intervention (P< 0.01); the difference after post-intervention was reduced significantly, compared with those in the pre-intervention (P<0.05). ③CMBC: The post-heating values and the difference in the experimental group were significantly higher than those in the control group (P<0.01). There were no statistical significant inner-group difference. Conclusion: Lower limb intermittent negative pressure therapy can improve the skin microcirculation of the quadriceps of the male rowers, which has a positive effect on the rapid recovery of physical fitness.
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    Research on the mechanism of high glucose affecting the apoptosis of schwann cells by Nox4 NADPH oxidase
    YU Ting , XIN Qing , XU Fei, LI Lei
    CJAP. 2019, 35 (2): 130-134.   DOI: 10.12047/j.cjap.5719.2019.029
    Abstract   PDF (1154KB) ( 198 )
    Objective: To investigate the mechanism of high glucose affecting the apoptosis of schwann cells through Nox4 NADPH oxidase. Methods: The schwann cells of newborn Wistar rats were cultured in vitro. The cultured cells were divided into four groups: control group, high-glucose group, NOX4 siRNA group and control siRNA group (n=10). The WST-1 method was used to detect the cell vitality, and the DCFH-DA method was used to detect the contents of intracellular reactive oxygen free radicals (ROS). Nox4 and Caspase3 mRNA expressions were detected by real-time fluorescence quantitative RT-PCR. Nox4 and Caspase3 protein expressions were determined by Western blot. Results: High glucose culture up-regulated Nox4 mRNA and protein expressions of schwann cells, decreased activity of schwann cells, increased intracellular ROS content, and promoted apoptosis by increasing Caspase3 mRNA and protein expressions. NOX4 siRNA blocked the accumulation of ROS in the high glucose cultured schwann cells, and reduced the damage of glucose on cell viability, by inhibiting NOX4 gene expression. NOX4 siRNA also reduced cell apoptosis by down-regulating Caspase3 mRNA and protein expressions. Conclusion: Nox4 was involved in the hyperglycemic-induced apoptosis of schwann cells through ROS. The regulation of Nox4 expression or function might be a new way to treat diabetic peripheral neuropathy.
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    Effects of prenatal maternal cold stress on spontaneous, exploratory and anxious behaviors in offspring
    LI Wen-jie, YANG Huan-min, LIAN Shuai, XU bin, WANG Li-peng, ZANG Shu-cheng, YUAN Jian-bin, DING Meng-yuan
    CJAP. 2019, 35 (2): 135-139.   DOI: 10.12047/j.cjap.5722.2019.030
    Abstract   PDF (2277KB) ( 191 )
    Objective: To study the effects of prenatal cold stress on the behavior and mood of offspring in pregnant rats. Methods: Six SPF-class Wister pregnant rats were randomly divided into normal temperature control group and cold stress group with 3 rats in each group. The pregnant female rats in the normal temperature control group were kept in the environment of (22±2)℃, and the pregnant female rats in the cold stress group were placed in the artificial intelligence climate chamber at(4±0.1)℃ for 7 days before the birth, and the young rats were divided into normal temperature after the young rats were born. After the young rats were born, they were divided into normal temperature control group of male rats (MR, 22), normal temperature control group of mother rats (FR, 15), cold stress group of male rats (MC, 15), and cold stress group of female rats (FC, 15) .In the fourth generation of the offspring, the open field experiment and the elevated cross maze test were carried out. Results: In the open field experiment, there was no significant difference in spontaneous activity and exploration behavior between the normal temperature control group and the cold stress group (P>0.05). In the elevated plus maze experiment, the retention time of the open arms, the number of open arms and the distance of the male and female rats in the cold stress group were significantly higher than those in the normal temperature control group (P<0.05). Conclusion: Prenatal maternal cold stress has no significant effect on spontaneous activity, exploration behavior and activity level of offspring, but the offspring have obvious abnormal behaviors with reduced anxiety behavior.
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    Treatment of type 2 diabetes mellitus with the stem and leaf of blacktartary buckwheat and its effects on the pancreas and spleen
    XIANG Wei-ling, JIN Li-qin, GAO Feng, XIAO Min, CHEN Yan
    CJAP. 2019, 35 (2): 140-144.   DOI: 10.12047/j.cjap.5670.2019.031
    Abstract   PDF (1241KB) ( 327 )
    Objective: To investigate the therapeutic effects of the black buckwheat leaf (BBL) in type 2 diabetes mellitus mice and its effects on pancreas and spleen. Methods: Forty male C57 / B16 mice (SPF) were randomly divided into normal control (NC) group (n=10) and the experimental group (n=30), the experimental group were fed with high sugar and high fat, combined with intraperitoneal injection of streptozotocin in small dose to establish the model of type 2 diabetes mellitus (T2DM). Those thirty model mice were randomly divided into 3 groups (n=10), diabetes mellitus group (DM), low dose of BBL (DM+L) treated group, high dose of BBL (DM+H) treated group. The mice in the NC group and the DM group were given normal saline per day, and the DM+L group and DM+H group were treated with black tartary buckwheat at the doses of 0.21g/kg·d-1 and 0.42g/kg·d-1 respectively. After 14 days. All mice were executed by cervical dislocation, then blood samples were collected, pancreas and spleen were removed for subsequent experiments. The serum levels of fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TCH) and insulin were detected. TNF-α protein in spleen tissue was detected by ELISA kit. The morphology of pancreas tissue was observed by HE staining, and the spleen coefficient was calculated. The expression levels of insulin receptor substrate-1(IRS-1) mRNA and IRS-1 protein in pancreatic tissue were detected. Results: Compared with the control group, the serum levels of FBG, TC and TCH in the model group were increased significantly, while the serum level of insulin was decreased significantly (P<0.05), the expression of TNF-α protein in spleen tissues was obviously raised, the expressions of IRS-1 mRNA and IRS-1 protein in pancreatic tissue in model group were decreased significantly (P<0.05). Compared with the model group, the serum levels of FBG, TC and TCH were decreased significantly in the BBL treated groups. The serum insulin level, spleen coefficient, TNF-α protein expression level in spleen tissue, IRS-1 mRNA expression and IRS-1 protein expression levels in pancreatic tissue in BBL treated group were increased significantly (P< 0.05). High-dose black tartary buckwheat leaves (0.42g/kg·d-1) exerted a more significant effect. Conclusion: Stem and leaf of black bitter buckwheat has significant therapeutic effects on reducing blood sugar and blood fat in type 2 diabetic mice, and has certain protective effects on pancreas and spleen of diabetic mice.
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    Intervention of curcumin and its analogue J7 on oxidative stress injury in testis of type 2 diabetic rats
    XU Fei-fei, MIAO Cheng-feng, CHI Chen, WU Gu, CHEN Guo-rong
    CJAP. 2019, 35 (2): 145-149.   DOI: 10.12047/j.cjap.5783.2019.032
    Abstract   PDF (2327KB) ( 247 )
    Objective: To investigate the intervention of curcumin and its analogue J7 on oxidative stress injury in testis of type 2 diabetic rats. Methods: Sixty male SD rats, 10 rats were chosen as normal control group (NC), the other 50 rats were assigned to experiment group. Experiment diabetic rats were induced by high-fat food and intraperitoneal injection of steptozotocin (STZ). After the model was established successfully, diabetic rats were divided into four groups randomly: diabetes mellitus group (DM, n=12), curcumin treatment group (CUR, n=10), high dose treatment group of J7 (J+, n=10), low dose treatment group of J7 (J-, n=10). The CUR group were intragastrically administered with curcumin 20 mg/kg daily, in addition, the J+ group and the J- group were intragastrically administered with J7 20 mg/kg and 10 mg/kg daily respectively. After 8 weeks, the fast blood glucose was detected biochemically. The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were detected by hydroxylamine method and thiobarbituric acid method respectively. The protein expressions of the nuclear factor-erythroid 2-related factor 2 (tNrf2), phosphorylation of Nrf2 (pNrf2), catalase (CAT), NAD(P)H quinine oxidoreductase 1 (NQO1) were measured by Western blot. The mRNA expressions of CAT, NQO1, hemeoxygenase-1 (HO1) were measured by quantitative real-time PCR (qRT-PCR). Morphological structure of testis was observed by hematoxylin-eosin (HE) staining. The expressions of Nrf2 and CAT were also detected by immunohistochemical method. Results: The levels of fast blood glucose and MDA in DM group were increased significantly(P<0.05), while the body weight, the activity of SOD, the protein expressions of pNrf2/tNrf2, CAT, NQO1 and the mRNA expressions of CAT, NQO1, HO1 were decreased (P<0.05). Under light microscope, the DM group showed disrupted histological appearance. Immunohistochemistry showed that the protein expressions of Nrf2 around the nucleus and CAT were decreased. With the treatment of curcumin and J7, the MDA levels in the three treatment groups were decreased (P<0.05). The activity of SOD, the protein expressions of pNrf2/tNrf2, CAT, NQO1 and the mRNA expressions of NQO1, HO1 were increased (P<0.05). the levels of fast blood glucose were decreased in the J+ and J- group (P<0.05), and the mRNA expression of CAT was increased in the J+ group (P<0.05). The ratio of pNrf2/tNrf2 in the J+ group was significantly higher than that in CUR and J- group (P<0.05). The protein level of CAT in the J+ group was also significantly higher than that in J- group (P<0.05). There were no significant differences in other indexes among the three treatment groups. Under light microscope, the morphology was obviously improved in the three treatment groups. Immunohistochemistry showed that the protein expressions of Nrf2 around the nucleus and CAT were increased in the three treatment groups. It was suggested that high dose J7 had better antioxidant stress ability in testis of diabetic rats. Conclusion: Curcumin and J7 could inhibit the oxidative stress damage of testicular tissue in diabetic rats, which might be related with the activation of the Nrf2-ARE signaling pathway.
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    Effects of aerobic exercise and glutamine on oxidative stress and expression of related factors in type 2 diabetic rats
    WANG Xiao-mei, JING Hui-feng
    CJAP. 2019, 35 (2): 150-154.   DOI: 10.12047/j.cjap.5777.2019.033
    Abstract   PDF (909KB) ( 227 )
    Objective: To investigate the effects of aerobic exercise and glutamine (Gln) on anti-oxidative stress and inflammatory factors in type 2 diabetes mellitus (T2MD) rats. Methods: Diabetic rat model was induced by streptozotocin (STZ). Fifty 6-week old male SD rats were randomly divided into 5 groups (n=10), including quiet control group (N), diabetes control group (D), diabetic aerobic exercise group (DE), diabetic glutamine group (DG) and diabetic aerobic exercise glutamine group (DEG). After 6 weeks, the related indicators of glucose and lipid metabolism, anti-oxidative stress and inflammatory factors in diabetic rats were detected, and the possible mechanism affecting inflammatory response were explored. Results: Compared with group N, the levels of serum malondialdehyde(MDA), blood glucose, total cholesterol(TC), triglyceride(TG), insulin, leptin and tumor necrosis factor-α(TNF-α) in group D were increased significantly (P<0.01). Compared with group D, serum levels of MDA, blood glucose, TC, TG, insulin, leptin and TNF-α in three intervention groups were decreased significantly, while the levels of SOD, GSH-Px and adiponectin were increased, and the combined effect was more obvious (P<0.01). Conclusion: Both aerobic exercise and Gln can relieve the glucose and lipid metabolism and disturbance, oxidative stress injury and inflammation in diabetic rats.
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    Selection of modeling time for type 2 diabetes mellitus mouse
    YANG Zhang-liang, TONG Hai-ling, SUN Meng-die, YUAN Jie, HU Ying, WANG Xu-tao, QI Min-you
    CJAP. 2019, 35 (2): 155-159.   DOI: 10.12047/j.cjap.5728.2019.034
    Abstract   PDF (1709KB) ( 466 )
    Objective: To analyze the changes of blood biochemical index and the pathological changes of myocardium and kidney in type 2 diabetic mouse at different time points, which can provide the basis for the selection of type 2 diabetic modeling time for later research. Methods: After 6 weeks of feeding with high-fat diet, 24 healthy male ICR mice were injected with streptozocin (STZ, 30 mg/kg) intraperitoneally for 5 days to establish diabetic models. After 9 days, a random blood glucose ≥ 11.1 mmol / L was measured as diabetic mice. 4, 6 and 8 weeks after successfully preparing the diabetic mouse, 8 diabetic mice (a group)would be sacrificed each time. Then the biochemical and pathological conditions were analyzed: ① the indexes of heart and kidney were calculated. ②the serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), creatinine (Cr) and blood urine nitrogen (BUN) were determined. ③ Histopathological changes of myocardium and renal tissues were observed by hematoxylin and eosin (HE) staining. Masson staining was used to observe the fibrosis of myocardium. PAS staining was adopted to observe the pathological changes of renal tissue. In addition, 8 ICR male mice were taken as the control group. Results: At the 4th, 6th and 8th week, cardiac organ coefficient, the values of LDH and CK were all increased compared with the control group. Cardiomyocyte hypertrophy and myocardial fibrosis could be observed. Renal organ coefficient, the values of Cr and BUN were increased. Glomerular hypertrophy, basement membrane thickening and atrophy could be perceived. Conclusion: At the 6th week, related biochemical and pathological changes in diabetic mice were comparatively obvious and breeding time was relatively short. Thus, 6 weeks after the preparation of the diabetic mice would be the optimal time for type 2 diabetes mellitus modeling, proper for inventions of drugs and other research purposes including pathology, physiology, biochemistry, etc.
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    Effects of high glucose induced primary cardiomyocytes injury on necroptosis and the related mechanism
    FANG Ting-ting, CAO Rui-ping, YE Hong-wei, MA Shan-feng, GAO Qin
    CJAP. 2019, 35 (2): 160-164.   DOI: 10.12047/j.cjap.5710.2019.035
    Abstract   PDF (2264KB) ( 262 )
    Objective: To observe whether necroptosis was happened in high glucose (HG) - induced primary cardiomyocytes injury and to investigate the likely mechanism. Methods: The primary cultured cardiomyocytes were divided into 4 groups (n=9): control group (the cardiomyocytes were incubated with 5.5 mmol/L glucose for 48 h), HG group (the cardiomyocytes were incubated with 30 mmol/L glucose for 48 h), HG + necrostatin-1 (Nec-1) group (the cardiomyocytes was co-incubated with necroptosis inhibitor Nec-1 at 100 μmol/L and HG for 48 h) and hypertonic pressure group (HPG, the cardiomyocytes was co-incubated with 5.5 mmol/L glucose and 24.5 mmol/L mannitol for 48 h). Cell viability was measured by MTT method, reactive oxygen species (ROS) generation was measured by DHE staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were tested by ELISA method. The mRNA and protein expressions of necroptosis related genes receptor interacting serine/threonine protein kinase 1 (RIP1), RIP3, mixed lineage kinase domain-like protein (MLKL) were tested by quantitative real-time PCR and Western blot. Results: The results showed HG intervention decreased cardiomyocytes viability, increased ROS generation, up-regulated the levels of TNF-α, IL-6 and IL-1β, increased RIP1, RIP3, MLKL expressions at mRNA and protein levels. Nec-1 treatment attenuated HG-induced increased cardiomyocytes viability, reduced ROS generation, down-regulated the levels of TNF-α, IL-6 and IL-1β, decreased RIP1, RIP3, MLKL expressions at mRNA and protein levels. Conclusion: Necroptosis was happened in high glucose-induced primary cardiomyocytes injury. Inhibition of necroptosis can reduce high glucose-induced cardiomyocytes damage, may be related to inhibition of oxidative stress and depression of inflammative factors releasing.
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    Effects of Notch signal on the expressions of HIF-α and autophagy- related genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation induced myocardial cell injury
    KONG Ling-yu , XI Zan , MA Wen-ting , YANG Fei-yun , NIU Li-dan , SHI Jin-he
    CJAP. 2019, 35 (2): 165-168.   DOI: 10.12047/j.cjap.5729.2019.036
    Abstract   PDF (1061KB) ( 389 )
    Objective: To investigate the effects of Notch signal on hypoxic induction factor (HIF-1α) and autophagy-associated genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation (OGD) induced myocardial cell injury. Methods: The OGD model was established using hypoxic culture box and hypoglycemic DMEM medium. The cells were divided into normal control group, OGD group, OGD + NC siRNA group, OGD + Notch1 siRNA group and OGD + HIF-1α siRNA group. Western blot was used to detect the interference effects of HIF-1α siRNA and Notch1 siRNA. The effects of Notch1 siRNA and HIF-1α siRNA on the activity of myocardial cells in OGD model were detected by the CCK-8 assay. The effects of Notch1 siRNA and HIF-1α siRNA on autophage-associated genes Beclin1, LC3I and LC3II expression were detected by Western blot. Results: The results of Western blot showed that HIF-1α siRNA could effectively knock down the expression of HIF-1α in myocardial cells in OGD model, and Notch1 siRNA could effectively knock down the expression of Notch1 and HIF-1α in myocardial cells in OGD model. The result of CCK-8 assay showed that Notch1 siRNA and HIF-1α siRNA reduced the activity of myocardial cells in OGD model, and there was no statistical difference between the two groups. Western blot results showed that Notch1 siRNA and HIF-1α siRNA could reduce the expressions of the autophagy-associated genes Beclin1, LC3I and LC3II, and reduce the ratio of LC3II to LC3I at mRNA level. Conclusion: Notch1 plays a role in myocardial protection by regulating the expression of HIF-1α to regulate the autophagy in OGD model cells.
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    The intervention of ginkgo biloba extract on monocrotaline-induced right ventricular hypertrophy in rats and its mechanism
    CHEN Hui-qin, CAI Ke-feng
    CJAP. 2019, 35 (2): 169-172.   DOI: 10.12047/j.cjap.5759.2019.037
    Abstract   PDF (5309KB) ( 207 )
    Objective: To study the protective effects of ginkgo biloba extract on the right ventricular hypertrophy. Methods: Seventy-two SD male rats were randomly divided into 3 groups: control group(CON), monocrotaline-induced right ventricular hypertrophy group (MCT) and ginkgo biloba extract treated group (EGB) (n=24 in each group). Group MCT and group EGB were intraperitoneally injected with 2%MCT at the dose of 60 mg /kg on the first day. From the second day, group MCT was injected with 2 ml 0.9% sodium chloride, and 60 mg/kg ginkgo leaf extract was administered to the stomach in group EGB. The control group was injected with 2 ml 0.9% sodium chloride on the first day. After 3 weeks, in each group,cardiac hemodynamic changes were measured, heart weight index was calculated, and myocardial pathological changes were observed by HE staining. The expression of TRPC6 was detected by real-time polymerase chain reaction (real-time -PCR) and Western blot. Results: Compared with the control group, the right ventricular systolic pressure (RVSP) was increased significantly in the MCT group(P<0.01), the maximum or decline rate of descent (RV±dp/dtmax) of the right ventricle pressure was increased significantly(P<0.01), while the EGB group had the same trend as all the indexes in the group MCT, but the amplitude of all indicators in group EGB were decreased significantly than those of group MCT(P<0.01), and the right ventricular hypertrophy index (RVMI) in group EGB was significantly lower than that in group MCT(P<0.01).Group MCT showed typical myocardial hypertrophy performance by HE staining, and the right ventricular myocytes in group EGB were significantly improved than that in group MCT, and the mRNA and protein expression levels of TRPC6 in the right ventricle of group MCT and group EGB were increased(P<0.01), while the EGB group was significantly lower than that of the MCT group(P<0.01). Conclusion: Ginkgo biloba extract may inhibit the signal pathway of CaN / NFAT in cardiac myocytes by reducing the expression of TRPC6 protein, and then play an early protective effect on myocardial hypertrophy.
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    Effects of simulated high-altitude hypobaric hypoxia on cardiac structure and function in rats
    FENG Zhen-long, ZHAO Tong, CHENG Xiang, ZHU Ling-ling , ZHAO Yong-qi, SHI Bing
    CJAP. 2019, 35 (2): 173-177.   DOI: 10.12047/j.cjap.5751.2019.038
    Abstract   PDF (5415KB) ( 333 )
    Objective: To investigate the effects of simulated hypobaric hypoxia environment at 7 000 m above sea level on cardiac structure and function in rats. Methods: A total of 96 male SD rats were randomly divided into high-altitude hypobaric hypoxia group (hypoxia group) and normobaric normoxia group (control group). Rats of hypoxia group were placed in a large cabin simulated 7 000 m high-altitude hypobaric hypoxia environment. Operating time 23 h / d, the control circadian ratio of approximately 12 h∶12 h. The rats in control group were bred under normobaric normoxia. The hypoxic group was divided into 3 d, 7 d, 14 d, 28 d groups according to hypoxic time, 12 rats in each group. Changes of structure and function of heart due to hypoxia were evaluated by echocardiography and electrocardiogram. Myocardial pathological changes were analyzed by hematoxylin-eosin staining(HE). Results: Compared with the control group at the same time point ①With prolonged exposure to hypobaric hypoxia, the growth ratio of body mass in rats is slower. Arterial oxygen saturation was significantly lower in both 14 d and 28 d (P<0.05). ② Left ventricular end-diastolic anterior wall thickness (LVAWD) and left ventricular end-diastolic posterior wall thickness (LVPWD) of rats in 28 d were increased significantly (P<0.05). Left ventricular end-diastolic diameter (LVIDD) and left ventricular internal dimension systole (LVIDS) of rats in 28 d were decreased significantly (P<0.05, P<0.01). Left ventricular ejection fraction (EF), fractional shortening of left ventricle (FS), pulmonary vein (PV) peak velocity and PV peak gradient of rats in 7 d were decreased significantly (P<0.05, P<0.01). ③The QRS and QT interval period were significantly prolonged in 14 d and 28 d (P<0.05, P<0.01). The ST was significantly lower in 3 d and 7 d (P<0.05, P<0.01). The amplitude of R wave gradually shifted downward in 7 d, 14 d, 28 d (P<0.05, P<0.01). ④The red blood cell (RBC), hemoglobin (HGB), red blood cell distribution width (RDW) in hypoxic group were increased significantly (P<0.01). The platelet count (PLT) count was decreased significantly in 14 d and 28 d (P<0.01). The serum creatinine (CR) was increased significantly in 14 d and 28 d (P<0.05). ⑤Pathological changes such as myocardial edema, sarcolemma condensate, focal degeneration and necrosis with inflammatory cell infiltration could be found at early stage of hypoxia. Myocardial compensatory repair such as myocardial fibroblasts proliferation was significant at end stage of hypoxia. Conclusion: Left ventricular systolic functions of rats were decreased significantly after exposure to high altitude hypoxia hypobaric. The left ventricular systolic functions would recovery compensatory after one week exposed to high altitude hypoxia hypobaric.
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    Effect of rutaecarpine on Alzheimer’s disease-like cognitive impairments induced by high glucose in rats
    CHEN Jian-guo, WU Ya-geng, LI Xiang
    CJAP. 2019, 35 (2): 178-182.   DOI: 10.12047/j.cjap.5805.2019.039
    Abstract   PDF (1816KB) ( 207 )
    Objective: To investigate the effects of rutaecarpine on high glucose-induced Alzheimer’s disease-like pathological and cognitive dysfunction and its mechanism in rats. Methods: Adult male SD rats were randomly divided into three groups (n=20): control group, high glucose group and rutaecarpine group. Rats in the control group were fed with conventional feed and tap water. The rats in the high glucose group were fed with conventional feed and 20% sucrose water. The rutaecarpine group was fed with fodder contain 0.01% rutaecarpine and 20% sucrose water. Morris water maze test was used to detect learning and memory and cognitive function of three groups rats after 24 weeks of feeding. Western blot analysis was used to detect tau protein at Thr205 and Ser214 sites in each group. Phosphorylation levels of GSK-3β in serine 9 site (S9-GSK-3β) and PP2A at cycline 307 site (Y307-PP2AC) were also detected. Immunohistochemistry further confirmed tau protein at Thr205 site in each group both in hippocampus and cortex. Results: Compared with the control group, Morris water maze results showed that the latency of finding the hidden platform of the rats in high glucose group was increased significantly and the number of crossing platforms and the target quadrant residence time were significantly decreased (all P<0.05). Immunohistochemistry showed that the phosphorylation level of tau protein at Thr205 site was significantly increased in the high glucose group compared with the control group, and the phosphorylation level of tau protein at Thr205 site in the rutaecarpine group was higher than that in the high glucose group. Western blot analysis showed that the phosphorylation level of tau protein in the high glucose group was significantly increased at Thr205 and Ser214 site compared with the control group, but the phosphorylation level of pS9-GSK-3β was significantly decreased (all P <0.05). Compared with the high glucose group, the latency of finding the hidden platform of the rats in rutaecarpine group was significantly decreased, and the number of crossing platforms and the target quadrant residence time were significantly increased (both P<0.05). Compared with the high glucose group, the phosphorylation levels of tau protein at Thr205 and Ser214 sites showed a significant decrease, but the phosphorylation level of pS9-GSK-3β was significantly increased (all P<0.05). Conclusion: Rutaecarpine can alleviate AD-like cognitive dysfunction induced by high glucose, possibly by enhancing pS9-GSK-3β phosphorylation, down-regulating GSK-3β activity, and thus reducing hyperphosphorylation of tau-associated sites.
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    Association between DIO polymorphism and aerobic endurance
    ZHOU Wen-ting, HU Yang
    CJAP. 2019, 35 (2): 183-188.   DOI: 10.12047/j.cjap.5733.2019.040
    Abstract   PDF (920KB) ( 195 )
    Objective: To study whether polymorphisms in the iodothyronine deiodinase (DIO) gene region contribute to endurance exercise capacity and to validate whether TSHR gene can be used as genetic marks associated with aerobic endurance performance. Methods: Three SNPs (C785T in DIO1 gene regions, Thr92Ala and Gly3Asp in DIO2 gene regions) were selected. The genotypes of the 123 elite long running athletes(EEA) and 127 college students from northern China(CG) were analyzed using the matrix assisted laser desorption ionisation-time of flight mass spectrometry method(MALDI-TOF). The athletes were divided into different groups according to the sports level and the items, which are international masters and masters (43 vs 80), 5/10 km and marathon (92 vs 31). Results: There was no significant difference of the C785T loci in DIO1 gene and the Thr92Ala loci in DIO2 gene between the EEA and the CG(P>0.05), while at Gly3Asp loci, the frequency distributions of the 3 genotypes were remarkably different in the groups of control and international masters of sports, as well as in the groups of control and marathon athletes(P<0.05). The genotype TT only existed in EEA not in CG, however, the frequency distribution was very low. The Thr92Ala and Gly3Asp loci of DIO2 gene were in strong linkage disequilibrium. The frequency distributions of the haplotype CT were significantly different in the male CG and the female CG, the male CG and the male EEA(P<0.05), the male CG and the male masters of sports, as well as in the male CG and the male marathon athletes(P<0.05). The frequency distributions of the haplotype TC were remarkably higher in the groups of female international masters of sports and female 5 000 m/10 000 m than those in the female CG(P<0.05). Conclusion: The frequency distributions of the haplotype CT were different in male and female CG, and haplotype CT could be used as a genetic mark associated with aerobic endurance performance of the male EEA, especially for the long running athletes of masters of sports and marathon, while the haplotype CT was associated with the aerobic endurance performance of the female long running athletes of international masters of sports and 5 000 m/10 000 m.
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    TECHNICAL NOTES
    A modified protocol for generating the simulated weightlessness rat model
    FU Zi-hao, WANG Zhen, WU Jie, YANG Hong-yan, ZHANG Xing, GAO Feng, LI Jia
    CJAP. 2019, 35 (2): 189-192.   DOI: 10.12047/j.cjap.5772.2019.041
    Abstract   PDF (4669KB) ( 216 )
    Objective: To introduce a modified protocol for generating the simulated weightlessness rat model by hindlimb unloading. Methods: Ninety male adult SD rats were randomly divided into three groups: the control group, classical suspension group and modified suspension group (n=30/group). In the classical suspension group, a strip of medical adhesive tape was attached to the tail, with horizontal filament tape wrapping. A piece of gauze was wrapped around the tail at the outermost layer and the tail was suspended for hindlimb unloading. In the modified suspension group, a layer of plastic net was added between the horizontal filament tape and the gauze to reduce the squeeze on the tail as a buffer zone and ensure proper circulation of the tail. After 4 weeks of suspension, damage to the tail and sheath detachment were observed. Meanwhile the body weight and right soleus wet weight of rats were measured. Results: The ratio of right soleus wet weight to body weight was decreased significantly in both the classical suspension group and the modified suspension group compared with the control group, while there was no difference in body weight among the three different groups. Importantly, the incidence of tail ischemia and necrosis (13.3% vs 40.0% in the classical suspension group) and the incidence of sheath detachment from tail (3.3% vs 26.7% in the classical suspension group) were significantly lower whereas the success rates of model (33.3% vs 83.3% in classical suspension group) was significantly higher in the modified suspension group. Conclusion: The modified protocol decreases the incidence of tail necrosis and sheath detachment in the rat tail suspension and increases the success rate of the hindlimb unloading rat model, with improved simplicity and practicability.
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  2019-03-15

Sponsor
Chinese Association for Physiological Sciences Academy of Military Medical Sciences Institute of Health and Environmental Medicine
Associate Sponsor
Institute of Basic Medical Sciences
Editor in Chief
WANG Hai
Edited and Published by
Editorial Board,Chinese Journal of Applide Physiology;Dali Dao,Tinanjin 300050,China



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