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  • Table of Content
      28 July 2020, Volume 36 Issue 4 Previous Issue    Next Issue
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    ORIGINAL ARTICLES
    Comparison of intermittent fasting and exercise on skeletal muscle autophagy activation and fat reduction
    WANG Zhen, YU Liang, FU Yue
    CJAP. 2020, 36 (4): 289-295.   DOI: 10.12047/j.cjap.5942.2020.063
    Abstract   PDF (2015KB) ( 458 )
    Objective: To observe the effect of weight control of intermittent fasting in different time course (14 d, 28 d)and explore its effect on skeletal muscle mass and autophagy. Methods: Sixty SD rats (male) were randomly divided into sedentary group (Sed), intermittent fasting group (InF) and exercise group (Exe), 20 rats of each group, intervention duration is 14 d and 28 d. Animals of InF group were fasted every other day, animals of Exe group underwent aerobic exercise on treadmill, the weight was recorded every week. The body fat mass was recorded by DEXA and then the fat mass index was calculated. The wet weight of bilateral soleus muscle was weighed and wet weight index was calculated. Immunofluorescence was used to detect laminin and light chain 3 (LC3), which reflect muscle fiber cross-sectional area and autophagosome respectively. Transmission electron microscopy was used to observe the number and morphology of autophagosomes. The expression levels of autophagy related proteins Unc-51 like kinase 1 (ULK1), LC3, sequestosome1 protein (p62), AMP activated protein kinase (AMPKα) and p-AMPKα (Thr172) were detected by Western blot. Results: ① From the 7th day of intervention, the body weight of rats in InF and Exe groups was significantly lower than that in Sed group, and the body weight of InF group was significantly lower than that in Exe group (P<0.01). After 28 days of intervention, the fat mass index in InF and Exe groups was significantly lower than that in Sed group, the fat mass index of InF group was significantly lower than that in Exe group (P<0.05).② After 28 days of intervention, the cross-sectional of muscle fibers in Exe group was significantly larger than that in Sed and InF groups (P<0.01). ③The expressions of AMPKα, p-AMPKα (Thr172) and ULK1 in InF and Exe groups were significantly higher than those in Sed group (P<0.05). However, at 14 days, only InF group showed the increase of LC3-II/LC3-I ratio and the decrease of p62 level (P<0.05). The same indicators of Exe group were only significant changed at 28 d. Conclusion: ① Intermittent fasting is superior to exercise in controlling the growth of body weight and body fat in rats. ② In terms of skeletal muscle autophagy activation, the length of intervention required for intermittent fasting (14 days) is shorter than aerobic exercise (28 days).
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    Effects of high-load exercise induced skeletal muscle injury on autophagy ultrastructure and Beclin1 and LC3-II / I in rats
    ZHANG Xin, WANG Rui-yuan
    CJAP. 2020, 36 (4): 296-300.   DOI: 10.12047/j.cjap.5944.2020.064
    Abstract   PDF (4053KB) ( 263 )
    Objective: To investigate the effects of high-load eccentric exercise on the ultrastructure of autophagy and the autophagy-related proteins Beclin1 and LC3II / I in rats. Methods: Forty-eight SD male rats were randomly divided into control group (C, n=8) and high-load eccentric exercise group (E, n=40) after adaptive training. Group E was run downhill for 90 minutes on the running platform, and soleus muscles were collected at 0, 12, 24, 48 and 72 hours after exercise. Transmission electron microscopy was used to observe the ultrastructural changes of skeletal muscle autophagosomes. Western blot was used to detect the expressions of Beclin1 and LC3II / I protein. Immunofluorescence was used to observe the localization and content of LC3. Results: The number of soleus muscle autophagosomes in group E was increased at 0, 12 and 24 hours after exercise, and LC3 autophagic fluorescence was significantly increased (P<0.01), while autophagic fluorescence at 48 hours after exercise was still increased significantly (P<0.05). Beclin1 and LC3II / I expression levels were increased after high-load centrifugal intervention (P<0.05), and were peaked at 12 h~24 h after exercise (P<0.01), and fully recovered at 72 h after exercise. Conclusion: High-load eccentric exercise can induce ultrastructural changes in skeletal muscle autophagy and increase the expression of autophagy protein. The peak value appears at 12 hours after exercise. The above may be one of the reasons for the decline in skeletal muscle function caused by sports injury.
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    Effects of aerobic exercise combined with Lycium ruthenicum on some indicators of myocardial lipid metabolism in rats with high-fat diet
    CUI Xiao-mei, CAO Jian-min, ZHANG Jing, ZHOU Hai-tao, NIU Yan-long, HU Ge, GUO Xian, LI Zhao-yan
    CJAP. 2020, 36 (4): 301-305.   DOI: 10.12047/j.cjap.5932.2020.065
    Abstract   PDF (952KB) ( 203 )
    Objective: To study the effects of aerobic exercise combined with Lycium ruthenicumon on some indicators of myocardial lipid metabolism in rats with high-fat diet. Methods: Fifty-five male Wistar rats were subjected to adaptive feeding for 4 days and weight-free swimming training for 3 days, 20 min/d. After eliminating 5 rats that were not suitable for swimming training, the others were randomly divided into 5 groups according to their weight: regular diet + quiet control group (RDC), high fat diet + quiet control group (HDC), high-fat diet + Lycium ruthenicum quiet control group (HDLC), high fat diet + aerobic exercise group (HDM), high fat diet + Lycium ruthenicum + aerobic exercise group (HDLM), 10 in each group. Group HDM and HDLM did 60 min/d swimming training for 6 weeks with no-bearing. Group C were fed regular diet; The other groups were fed with high-fat diet; Group HDLC and HDLM were intragastrically treated with Lycium ruthenicum at the dose of 4.48 g/(kg·d), and the volume was 5 mL/kg, and the other groups were given equivalent distilled water. The Lee's index, serum and myocardial biochemical indexes were measured after 6 weeks. Results: Compared with group RDC, Lee's index, serum free fatty acids (FFA), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), myocardial FFA and intercellular adhesion molecule-1 (ICAM-1) increased significantly (P<0.01), serum level of high-density lipoprotein cholesterol (HDL-C) decreased significantly (P<0.01) in group HDC. Compared with group HDC, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-C, myocardial FFA and ICAM-1 decreased significantly (P<0.05 or P<0.01), serum HDL-C levels increased significantly (P<0.05 or P<0.01) in group HDLC, HDM and HDLM. Compared with group HDLC and HDM, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-C, myocardial FFA and ICAM-1 decreased significantly (P<0.05), serum HDL-C level increased significantly (P<0.05) in group HDLM. Conclusion: Aerobic exercise and/or Lycium ruthenicum can improve lipid metabolism in rats with high-fat diet, reduce lipotoxicity caused by obesity. Combined intervention is more effective.
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    Effects of acorus tatarinowii Schott and alpha asarone on free radicals and nNOS/NO in hippocampus of rats with fatigue movement
    ZHU Mei-ju, MAO Ze-Hua, GUO Hong-ying, ZHU Hong-zhu, DING Xiao-min
    CJAP. 2020, 36 (4): 306-311.   DOI: 10.12047/j.cjap.5947.2020.066
    Abstract   PDF (931KB) ( 135 )
    Objective: To study the effects of acorus tatarinowii Schott and its active ingredient-alpha-asarone on learning and memory, free radical metabolism and nNOS/NO signal in hippocampus of rats with fatigue movement. Methods: Eighty SD male rats were randomly divided into eight groups: control group(A), exercise group(B), exercise + alpha-asarone low, middle and high dose treatment group (C, D, E), exercise + acorus tatarinowii Schott low, middle and high dose treatment group (F, G, H),with ten rats in each group. The rats in group C, D and E were administered with alpha-asarone at the doses of 0.10, 0.50 and 1.00 mg.kg-1.WT-1 by ig. The rats in group F, G and H were administered with the extracts of Acorus tatarinowii Schott of at the doses of 0.12, 1.20 and 4.80 g.kg-1.WT-1 by ig. Learning and memory of rats were tested by the method of water maze experiment, and the activities of SOD and NOS, the contents of MDA were detected by the biochemical methods, and the expression levels of nNOS protein in hippocampus of rats were tested by the method of Western blot in at the end of the experiment. Results: The escape latency and MDA content in hippocampus of rats in groups E and H were lower than those in groups B, C, D, F and G and the numbers of Plateau crossing, SOD and NOS activities and the expression levels of nNOS protein in hippocampus of rats were higher than those in groups B, C, D, F and G(P<0.01). The activities of SOD in hippocampus of rats in groups A, E and H were A>E>H, whereas the contents of MDA were opposite (P<0.01); the activities of NOS and the expression levels of nNOS protein in hippocampus of group E were lower than those of groups A and H (P<0.01 or P<0.05), but there was no significant difference between groups A and H (P>0.05). There were no significant difference in escape latency and numbers of crossing platform among groups A, E and H (P>0.05). Conclusion: Acorus tatarinowii Schott and alpha-asarone can significantly improve learning and memory of rats with fatigue movement. The mechanism is related to reclaiming the imbalance of free radical metabolism and up-regulating nNOS/NO signal in hippocampus of the rats.
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    Effects of aerobic exercise on PPARα signaling in diabetes rats and its association with PPARγ
    WANG Tian-yuan, WANG Xiao-hui
    CJAP. 2020, 36 (4): 312-317.   DOI: 10.12047/j.cjap.5945.2020.067
    Abstract   PDF (1484KB) ( 368 )
    Objective: To investigate the improved effects of 4-week aerobic exercise on blood glucose and lipid of diabetes mellitus (DM) rats and its association with peroxisome proliferators-activated receptor (PPAR)α signaling and PPARγ. Methods: Six-week old male SD rats were fed with 8-week high-fat diet following a single intraperitoneal injection of streptozotocin to establish DM models rats. Except control group rats (fed with ordinary diet), DM model rats were randomly divided into DM group, trained DM group (TDM), TDM plus PPARγ agonist pioglitazone group (TDP), and TDM plus PPARγ inhibitor GW9662 group (TDG), 8 rats in each group. TDM, TDP and TDG rats were undertaken a medium-intensity incremental treadmill training for 4 weeks, once a day and 6 days a week (1st week: 15 m/min speed lasting for 30 min; 2nd week: 15 m /min for 60 min; 3rd week: 20 m/min for 60 min; 4th week: 20 m/min for 90 min). All the rats were fed with ordinary diet during 4-week exercise. The rats were anesthetized and blood samples were collected at 36 h after the last exercise, and liver and gastrocnemius were collected after the rats were sacrificed. Fasting blood glucose and blood lipid were detected (blood glucose, insulin and TC, TG, HDL and LDL). The protein levels of PPARα, adenosine monophosphate (AMP)-activated protein kinase (AMPK) and carnitine palmitoyl transferase 1 (CPT1) in the liver and gastrocnemius of the rats were measured by Western blot. Results: ①Compared with control rats, the serum levels of TC, TG, LDL, and FBG (>11.1 mmol/L) in DM rats were significantly increased, indicated the successful establishment of DM rats. ②Compared with DM rats, accompanied with the improvements of fasting blood glucose and blood lipid, the protein levels of PPARα, AMPK and CPT1 were significantly increased in the liver and gastrocnemius of TDM rats. ③Compared with TDM rats, no significant changes were found in TDG rats including the protein levels of PPARα and CPT1 (liver and gastrocnemius) as well as AMPK (liver), except for a decrease of AMPK in gastrocnemius; while the protein levels of PPARα, AMPK and CPT1 were all increased in the liver and gastrocnemius of TDP rats. Conclusion: Aerobic exercise-induced improvements of blood glucose and blood lipid was related to the up-regulation of AMPK-PPARα-CPT1 signaling pathway in the liver and muscle of DM rats. The exercise-induced enhancement of PPARα in DM rats was unrelated to PPARγ, but PPARγ activation could further increase the protein levels of AMPK-PPARα-CPT1 in TDM rats.
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    Effects of exogenous H2S on hepatic fibrosis in diabetic mice and its mechanism
    XI Yu-xin, WEN Xin, JIAO Li-jie, WEI Ya-xin, CHANG Gui-quan, WU Ren, SUN Feng-qi, HAO Jing-hui, LI Hong-zhu
    CJAP. 2020, 36 (4): 318-320.   DOI: 10.12047/j.cjap.5965.2020.068
    Abstract   PDF (1197KB) ( 226 )
    Objective: To investigate the effects of exogenous hydrogen sulfide (H2S) on the hepatic fibrosis in diabetic mice and its mechanism. Methods: Twenty-four C57 male mice (weight 22±2 g) were randomly divided into three groups (n=8): ① Normal control group (Control): Mice were intraperitoneally injected equal amount of normal saline, the injection time was the same as that of the experimental groups; ② Diabetes model groups (HG): Streptozotocin (STZ) was injected intraperitoneally once according to body weight (150 mg/kg) to establish diabetes model; ③ NaHS treatment groups (HG + NaHS): Mice were intraperitoneally injected with NaHS (100 μmol/L·kg·d) once a day for 12 consecutive weeks. The hepatocyte injury was detected by HE staining; the hepatic fibrosis was observed through Masson staining; the protein expressions of cystathionine - β - synthetase (CBS), collagen-I (CoL-I), collagen-III (CoL-III) and matrix metalloproteinase-9 (MMP-9) were detected by Western blot. Results: Compared with the control group, the damage and fibrosis of hepatocyte were significantly aggravated, the expression of CBS proteins was decreased (P<0.01), and the expression levels of CoL-I, CoL-III and MMP-9 proteins were increased (P<0.01) in the diabetic model group. Compared with the diabetic model group, the damage and fibrosis of hepatocyte were significantly lightened, the expression of CBS proteins was obviously increased (P<0.01), and the expression levels of CoL-I, CoL-III and MMP-9 proteins were markedly decreased (P< 0.01). Conclusion: H2S inhibits the hepatic fibrosis in diabetic mice, and its mechanism is related to the decrease of collagen and matrix metalloproteinase-9.
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    BRIEF REPORTS
    Effects of long-term Tai Chi exercise on knee joints skin microvascular reactivity and transcutaneous oxygen partial pressure in middle and old aged people
    ZHU Huan, HU Qing-hua, PENG Ai-ping, WAN Li
    CJAP. 2020, 36 (4): 321-323.   DOI: 10.12047/j.cjap.5957.2020.069
    Abstract   PDF (906KB) ( 194 )
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    ORIGINAL ARTICLES
    Expression of thioredoxin reductase 1 (TrxR1) in gastric cancer and its effect on the growth of gastric cancer cells
    FAN You-shou, QIU Zhi-sheng, ZHAO Ya-ping
    CJAP. 2020, 36 (4): 324-329.   DOI: 10.12047/j.cjap.5967.2020.070
    Abstract   PDF (999KB) ( 160 )
    Objective: To investigate the relationship between the expression of thioredoxin reductase 1 (TrxR1) in gastric cancer tissues and the survival time of patients with gastric cancer and its effect on the growth of gastric cancer cells. Methods: The expression level of TrxR1 mRNA in gastric cancer tissues and adjacent tissues of 76 patients was determined by real-time PCR assays, and the relationship between the mRNA expression level of TrxR1 and the clinicopathological characteristics and prognosis of gastric cancer patients was analyzed. Three gastric cancer tissues and adjacent tissues were randomly selected, and the expression of TrxR1 protein was detected by immunohistochemistry and Western blot. The expression levels of TrxR1 in human gastric cancer cells line and gastric mucosa epithelial cells were determined by Western blot and quantitative real-time PCR assays. Then, AGS cells were transfected with siRNA sequences to silence the expression of TrxR1. AGS cells were divided into 3 groups according to different processing negative control group: transfected with NC-siRNA, TRXR1 siRNA interference group 1: transfected with TRXR1-siRNA1, TRXR1 siRNA interference 2 group: transfected with TRXR1-siRNA2. The expression of TrxR1 mRNA in AGS cells of each group was detected by Real-time PCR. The proliferation of AGS cells was determined by MTT and colony formation assays following TrxR1 silencing. Results: TrxR1 mRNA and protein expression levels in gastric cancer tissues were significantly up-regulated compared with adjacent tissues, and were mainly located in the cytoplasm. High expression of TrxR1 was associated with TNM stage and lymph node metastasis, and the overall survival time of patients with high expression of TrxR1 was shorter than those with low expression level. TrxR1 mRNA and protein in AGS cells of TRXR1-siRNA1 group and AGS cells of TRXR1-siRNA2 group were significantly reduced compared with NC-siRNA group (P<0.05). And AGS cell clone formation and proliferation ability were decreased. Conclusion: The high expression of TrxR1 in gastric cancer tissues indicates poor prognosis of patients, and the silencing of TrxR1 can inhibit the proliferation of gastric cancer cells.
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    Effects of a novel polyamine metabolic enzyme inhibitor SI-4650 on proliferation of colon cancer CT-26 cells and its mechanism
    HUANG Jiao-jiao, WANG Yan-lin, SUN Li-dan, CAO Chun-yu, QIN Ye, YANG Jian-lin
    CJAP. 2020, 36 (4): 330-335.   DOI: 10.12047/j.cjap.5968.2020.071
    Abstract   PDF (1117KB) ( 262 )
    Objective: To investigate the effects of a novel polyamine metabolism enzyme inhibitor SI-4650 on autophagy and apoptosis of colon cancer CT-26 cells as well as their correlation. Methods: CT-26 cells treated with 40, 80 μmol·L-1 SI-4650 alone or in combination with 3-MA were used as experimental group. CT-26 cells treated with 0 μmol·L-1 SI-4650 alone or in combination with 3-MA were used as control group. Chemiluminescence was used to analyze the effect of SI-4650 on spermine oxidase (SMO) and acetylpolyamine oxidase(APAO) activity. High performance liquid chromatography (HPLC) was performed to detect cellular polyamine content.The CCK8 method was used to detect the inhibitory effect of SI-4650 on proliferation of CT-26 cells. PI single-staining/flow cytometry (FCM) were used to analyze cell cycle. Western blot were used to analyze autophagy. Apoptosis was analyzed by PI/FITC-Annexin V double staining, JC-1 fluorescent probe and Fluo-3 AM calcium ion fluorescent probe combined with flow cytometry and Western blot. Results: CCK8 assay showed that 24-,48-,72-hours treated with SI-4650 all could inhibit the proliferative activity of CT-26 cells in a dose- and time-dependent manner (P<0.01) . The inhibition rate was 36.98% and 46.91% in 40 μmol·L-1 SI-4650 group and 80 μmol·L-1 SI-4650 group respectively. SI-4650 could significantly inhibit the activities of SMO and APAO interfere with polyamine metabolism and reduce the content of total polyamine in CT-26 cells (P<0.01). SI-4650 could block CT-26 cells in G0/G1 phase, significantly reduce the number of cells in S phase(P<0.01), and lead to a significant increase in the contents of autophagy-related Beclin-1, LC3-II in CT-26 cells(P<0.01); At the same time, the concentration of calcium in CT-26 cells was increased, the mitochondrial membrane potential was decreased, the expressions of c-PARP and Bax were increased, the content of Bcl-2 was decreased, and the number of apoptotic cells was increased. After SI-4650 combined with autophagy inhibitor 3-MA treatment of CT-26 cells, the level of autophagy, the apoptosis-related protein, mitochondrial membrane potential and calcium ion concentration were decreased, and the number of apoptotic cells was decreased. Conclusion: SI-4650 has the pharmacological activity of killing colon cancer CT-26 cells, and its mechanism may be related to the interference of polyamine metabolism and induction of cell apoptosis and autophagy. In this process, autophagy is inhibited to block apoptosis, autophagy and apoptosis combined to kill tumor cells.
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    Effect of cyclic GMP-AMP synthase on EMT in breast cancer cells
    ZHANG Xiao-jin, ZHANG Wen-xia, ZHU Ji-ling, LUO Wen-pan
    CJAP. 2020, 36 (4): 336-339.   DOI: 10.12047/j.cjap.5917.2020.072
    Abstract   PDF (1349KB) ( 144 )
    Objective: To investigate the effects of cyclic GMP-AMP synthase (cGAS) on the proliferation, migration and epithelial to mesenchymal transition (EMT) of breast cancer cells. Methods: The cGAS lentiviral vector and control fluorescence vector were transfected into breast cancer MCF7 cells and were divided into negative group (NC) and MCF7-cGAS group. The effect of cGAS on proliferation in the MCF7 cells was detected by MTT. The effect of cGAS on cell migration was detected by Transwell assay. The expressions of EMT related proteins were analyzed by Western blot. Results: After over-expressed with cGAS, the proliferation and migration of MCF7 cells were increased (P<0.05). The expression level of the epithelial markers E-cadherin was decreased, while the expression level of the mesenchymal markers N-cadherin was increased(P<0.05). Conclusion: The over-expression of cGAS increased the proliferation and migration of breast cancer cells and induced EMT in breast cancer cells.
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    miR-520a-3p regulates the signaling pathway of cytokine secretion in cervical cancer
    LIU Hong-yan, LU Qi-hong, HUANG Chu-ying
    CJAP. 2020, 36 (4): 340-345.   DOI: 10.12047/j.cjap.5935.2020.073
    Abstract   PDF (1122KB) ( 280 )
    Objective: To investigate the molecular mechanism of miR-520a-3p regulating the secretion of cytokines in cervical cancer. Methods: The matching between miR-520a-3p and the NF-κB complex subunit RELA was analyzed by TargetScanHuman, and then the luciferase reporting system was used to detect whether miR-520a-3p targeted the NF-κB complex subunit RELA. After cervical cancer HELA cells stimulated by LPS, in the overexpression group or the knockout group, miR-520a-3p mimics or inhibitor were mixed with transfection reagent and dripped into HELA cells. The expression levels of colony stimulating factor (GCSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 19 (IL-9), interleukin 10 (IL-10), interleukin 12 p40 (IL-12 p40), interleukin 12 p70 (IL-12 p70), interleukin 13 (IL-13), interleukin 17 (IL-17), interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), monocyte chemoattractant protein-5 (MCP-5), small inducible cytokine A5 (RANTES), tumor necrosis factor-alpha (TNFα) were detected by enzyme-linked immunosorbent assay in both the overexpression group and the knockout group. Each experiment was repeated three times. Results: miR-520a-3p targeted the 3’UTR of RELA. After activation of the NF-κB signaling pathway by lipopolysaccharide (LPS), the protein expression levels of cytokines GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα secreted by HELA cells of cervical cancer were increased significantly (P<0.05). In the overexpression group, the protein expression level of RELA of NF-κB complex subunit was decreased, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were decreased (P<0.05). In the knockdown group, the protein expression level of RELA, a subunit of NF-κB complex, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were increased (P<0.05). Conclusion: miR-520a-3p inhibits cytokine secretion in HELA cells of cervical cancer by targeting RELA, a key molecule in the NF-κB signaling pathway.
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    Astragalus injection mediates autophagy and myocardial remodeling in rats with ischemic cardiomyopathy through calumenin
    SHAO Li-qun, BAI Ting-ting, ZHAO Qiang, WANG Chun-gui, XUAN Li-ying, WANG Yu, ZHAO Ming
    CJAP. 2020, 36 (4): 346-349.   DOI: 10.12047/j.cjap.5922.2020.074
    Abstract   PDF (1785KB) ( 191 )
    Objective: To study the effects of astragalus injection on myocardial remodeling, calumenin and autophagy in rats with ischemic cardiomyopathy.Methods: Thirty-six male SD rats were divided into normal control group, ischemic cardiomyopathy group and astragalus injection group, 12 in each group. Electrocardiogram (ECG) and echocardiography were performed before operation in three groups. Rats in ischemic cardiomyopathy group and astragalus injection group underwent thoracotomy and ligation of coronary artery for 20 minutes, then thoracic cavity was closed after reperfusion. In the astragalus injection group,10 g/kg body weight of Astragalus injection was injected once a week, four times in total. Four weeks after operation, rats in three groups were executed by echocardiography and their hearts were collected for Hematoxylin-Eosin (HE) staining and Van Gieson (VG) staining to observe myocardial pathological changes. Calumenin, LC3-I, LC3-II expressions and LC3-I/LC3-II ratio were detected by Western blot.Results: Compared with ischemic cardiomyopathy group, the echocardiography and myocardial pathology of rats in astragalus injection group changed obviously, and the expressions of calumenin, LC3-I, LC3-II and LC3-I/LC3-II ratio changed significantly (P<0.01).Conclusion: Astragalus injection has apparent inhibitory effect on ventricular remodeling and autophagy of myocardial cells in rats with ischemic cardiomyopathy, which may be mediated by calumenin.
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    Effects of oxymatrine and vincristine on drug resistance in HCT-8/VCR cells and its mechanism
    SU Jian-wei, ZHOU Xi-han, YE Ying-xia, JIANG Qi
    CJAP. 2020, 36 (4): 350-353.   DOI: 10.12047/j.cjap.5925.2020.075
    Abstract   PDF (1126KB) ( 205 )
    Objective: To study the effects of oxymatrine and vincristine on resistance in HCT-8/VCR cells and its mechanism. Methods: HCT-8 / VCR cells were cultured in vitro and were divided into blank control group, oxymatrine group, vincristine group, oxymatrine and vincristine combined group, each group had 6 complexes. The drug resistance of HCT-8/VCR cells was investigated by CCK-8 when treated with vincristine alone or in combination with oxymatrine. The autophagy was determined by monodansylcadaverine (MDC) staining. The level of IL-6 was detected by ELISA. The expressions of autophagy-related gene P62, LC3-Ⅱ / LC3-Ⅰ, Beclin-1 protein and TLR4 were detected by Western blot assay. Results: Oxymatrine combined with vincristine could reduce the drug resistance of HCT-8 / VCR cells by the reversal multiple of 3.23. Compared with the blank control group, the content of autophagosome and the content of IL-6 in the oxymatrine group and the combination group were also decreased significantly (P<0.01). The content of autophagosome in the vincristine group was increased and the content of IL-6 was also significantly increased (P<0.01). Compared with the oxymatrine group, the combination group had higher autophagosome content, while IL-6 content was decreased (P<0.01); Western blot experiments showed that compared with the blank control group, the expression of P62 in the oxymatrine group was decreased (P<0.05), while the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1 and TLR4 were all increased (P<0.05). The expression of P62 in the vincristine group and the combined group was increased (P<0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (P<0.05). Compared with the vincristine group, the expression of P62 was increased in the combination group (P<0.05), and the expressions of LC3-Ⅱ / LC3-Ⅰ, Beclin-1, and TLR4 were all decreased (P<0.05). Conclusion: Oxymatrine combined with vincristine can reduce the drug resistance of HCT-8/VCR cells, which may be related to the regulation of autophagy activity and TLR4 signal activation.
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    Effects of modified Xiaoyao San on TLR4/NF-κB pathway in hippocampal microglia of LPS-induced depression model rats
    GUO Rui, QIN Wei-shuai, ZHANG Shuang-yong, ZHANG Shu-ling
    CJAP. 2020, 36 (4): 354-357.   DOI: 10.12047/j.cjap.5936.2020.076
    Abstract   PDF (1092KB) ( 278 )
    Objective: To investigate the effects of modified Xiaoyao San on TLR4/NF-κB pathway in hippocampal microglia of LPS-induced depression model rats, and to explore its antidepressant mechanism. Methods: SD rats were randomly divided into control, model, fluoxetine (10 mg·kg-1), low and high dose of modified Xiaoyao San (3.64, 7.28g·kg-1) group. The depression model was established by chronic LPS injection (ip, 0.5 mg·kg-1) and rats were treated by intragastric administration for 14 days. After the model was established, the depression-like behavior of rats was evaluated by open field and forced swimming test. The expression of microglia marker protein Iba-1 was detected by immunohistochemistry. The levels of TNF-α and IL-6 in hippocampal homogenate were detected by ELISA method and the expressions of TLR4 and NF-κB protein in hippocampus were detected by Western blot. Results: Compared with control group, the depression-like behavior was significant in model group rats (P<0.01), the microglia in the brain was activated (P<0.01), the contents of TNF-α and IL-6 in the hippocampus were increased (P<0.01), and the expression levels of TLR4 and NF-κB proteins were up-regulated (P<0.01). Compared with model group, the depression-like behavior of the rats in the fluoxetine and high-dose modified Xiaoyao San group was significantly alleviated (P<0.05), the expression of Iba-1 in microglia returned to normal (P<0.01), the contents of TNF-α and IL-6 were decreased (P<0.01), and the expression levels of TLR4 and NF-κB protein were decreased (P<0.05). Compared with fluoxetine group, the high-dose modified Xiaoyao San group had no statistically significant difference in each index, suggesting that there was no significant difference in the antidepressant effect between the two groups. Conclusion: Modified Xiaoyao San can significantly improve the depression-like behavior in rats, and its mechanism may be related to inhibiting the TLR4/NF-κB pathway of microglia and down-regulating the expression of inflammatory factors.
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    Expression of ANO1 during cardiac fibroblasts differentiation and its electrophysiological characteristics as calcium-activated chloride channel protein
    TIAN Xiang-qin, TAN Chao-yang, LI Na-na, CHANG Yu-qiao, ZHANG Ai-mei, ZHOU Ying, MA Ke-tao, SI Jun-qiang
    CJAP. 2020, 36 (4): 358-362.   DOI: 10.12047/j.cjap.5940.2020.077
    Abstract   PDF (4711KB) ( 148 )
    Objective: To investigate the expression and electrophysiological characteristics of calcium-activated chlorine channel anoctamin-1 (ANO1) protein during the differentiation of cardiac fibroblasts (CFs) into myofibroblasts (MFs), and to elucidate the role of ANO1 in myocardial fibrosis. Methods: The primary CFs from neonatal rats were isolated and the cells differentiated into MFs by subculture. The Ca2+-activated Cl- current (ICl(Ca)) in CFs and MFs were measured by whole-cell patch clamp, and the expressions of ANO1, α-smooth muscle actin(α-SMA)and vimentin in CFs and MFs were detected by immunofluorescence assay and Western blot, respectively. Results: The current density in the early adherent CFs was stronger than that in MFs. ANO1 was expressed preferentially within and around the nuclei, and a small amount of ANO1 was expressed on the cell membrane. Moreover, ANO1 expression was weak in the early adherent CFs and displayed stronger expression in the MFs with proliferation tendency. Conclusion: The expression of ANO1 is closely related to the differentiation of MFs and it may be involved in modulation myocardial fibrosis.
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    Expression of Sirtuin 1 in visceral adipose tissue in Tibetan mini-pigs with obesity and insulin resistance induced by high fat/cholesterol diet
    RONG Yi-li, PAN Yong-ming, HUANG Jun-jie, YU Chen, ZHU Ke-yan, CHEN Min-li
    CJAP. 2020, 36 (4): 363-368.   DOI: 10.12047/j.cjap.5977.2020.078
    Abstract   PDF (961KB) ( 335 )
    Objective: To investigate the role of Sirt1 in visceral adipose tissue in Tibetan mini-pigs with obesity and insulin resistance induced by high fat/cholesterol diet. Methods: Twelve male Tibetan mini-pigs were divided into 2 groups randomly: normal control (NC) group, high-fat/cholesterol (HFC) diet group, 6 in each group. After 16 weeks of modeling, fasting body weight and body mass index (BMI) were measured. Total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were measured in anterior venous blood, and atherosclerosis index (AI) was calculated. Meanwhile, intravenous glucose tolerance test was conducted to observe the changes of blood glucose and insulin, and the area under the curve (AUC) was calculated. After euthanasia, visceral fat rate was detected, and visceral fat tissue was taken for histopathological observation and fat cell diameter analysis. RT-PCR was used to observe the mRNA expression levels of Sirtuin1 (Sirt1), insulin-like growth factor-1 (IGF-1), glucose transporter 4 (GLUT4), peroxisome proliferator-activated receptor γ (PPARγ), peroxisome proliferator-activated receptor γ-assisted activator 1α (PGC-1α), forkhead box protein O1 (FoxO1), lipolysis-related gene hormone-sensitive lipase (HSL), and fat synthesis-related gene fatty acid synthase (FASN)changes in adipose tissue. Results: Compared with the NC group, the body weight, BMI, TC, LDL-C, HDL-C, AI and visceral fat rate were significantly increased after 16 weeks of high-fat/cholesterol induction in Tibetan mini-pigs(P<0.05,P<0.01). Meanwhile, the glucose tolerance curve was significantly delayed and the area under the curve of blood glucose and insulin was significantly increased (P<0.05). HE pathological observation and quantitative analysis showed that fat cells were hypertrophy and the average cell diameter was increased significantly (P<0.01). In addition, the mRNA expression levels of Sirt1,PGC-1α, GLUT4, and HSL were all decreased in varying degrees in adipose tissue, among which the mRNA expressions of Sirt1 and HSL were significantly decreased (P<0.05), while the mRNA expressions of FOXO1, IGF-1, PPARγ, and FASN were significantly increased (P<0.05, P<0.01). Conclusion: Tibetan mini-pigs were induced by high fat/cholesterol diet to form obesity model with phenotypic characteristics such as lipid disorder and insulin resistance, whereas Sirt1 plays a key role in visceral fat deposition and insulin sensitivity reduction in obese Tibetan mini-pigs.
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    Thrombolytic effects of recombinant staphylokinase on coronary thrombosis in miniature pigs
    LI Guang, FANG Xiao-Lan, REN Zhong, LI Ke-qiang, ZHOU Song-hui, HU Dao-qi, WU Gang-qiang, WANG Tai-lin, LI Wei-ping
    CJAP. 2020, 36 (4): 369-374.   DOI: 10.12047/j.cjap.5994.2020.079
    Abstract   PDF (971KB) ( 148 )
    Objective: To evaluate the thrombolytic effects of recombinant staphylokinase and compare it with those of recombinant streptokinase. Methods: Thirty Chinese experimental miniature pigs were divided into five groups, namely, solvent control group, positive drug control group and three recombinant staphylokinase groups, six in each group. The thrombus of coronary artery was formed by surgical thoracotomy and direct current stimulation in anesthetized animals. Intravenous administration was started after the thrombus of coronary artery was formed for 30 minutes, and the method of first injection and then constant speed infusion by peristaltic pump was used. The solvent control group was injected intravenously with solvent, the positive drug control group was given recombinant streptokinase 4 mg·kg-1 intravenously, and the three recombinant staphylokinase groups were given recombinant staphylokinase at the doses of 4, 2 and 1 mg.kg-1 intravenously. The volume of intravenous injection was 5 ml, which was completed within 1 min, the speed of infusion was 0.5 ml·min-1, which was completed within 60 min, and the animals were sacrificed 120 minutes later. Before and 30, 60 and 120 min after administration, the venous blood samples were collected. At the end of the experiment, the coronary artery segments of the thrombosis site were taken, and the euglobulin dissolution time (ELT), blood fibrinogen content (FBG), fibrinogen degradation product (FDP) and wound bleeding volume were measured respectively. The coronary thrombolysis rate, myocardial ischemia degree and ischemia range were measured.Results: Compared with the solvent control group, ELT in the experimental group was significantly shortened (P<0.05 or P<0.01), FBG degradation in a few experimental animals was more than 20%, FDP was significantly increased (P<0.05 or P<0.01), and there was no significant effect on blood pressure and heart rate of small pigs. Compared with the control group, the maximum thrombus area was decreased by 34.3% and 15.4% (P<0.05) in the high and middle dose groups of the experimental group. Compared with the same dose of recombinant streptokinase, recombinant staphylokinase had stronger thrombolytic effect (P<0.05 or P<0.01) on the coronary thrombus caused by electrical stimulation, less bleeding side effects and the same effect on the degree and range of myocardial infarction as recombinant streptokinase. Conclusion: Compared with recombinant streptokinase, recombinant staphylokinase has faster thrombolysis speed, higher fibrin specificity and less bleeding side effects. In general, 2 mg.kg-1 recombinant staphylokinase has better efficacy and safety.
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    BRIEF REPORTS
    p-Coumaric acid ameliorates chronic restraint stress induced memory deficit through BDNF in mice
    XIAO Chu-li, XIE Yi-ming, LI Jin-cheng, PENG Li-wen, XIAO Zhi-yong, YU Xu-dong
    CJAP. 2020, 36 (4): 375-378.   DOI: 10.12047/j.cjap.5874.2020.080
    Abstract   PDF (941KB) ( 170 )
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    TECHNICAL NOTES
    Quantitative analysis of the effects of different stimuli on the contraction of gastrocnemius in vivo and in vitro specimen
    XU Pei-hong, TANG Feng, WANG Xue-wei, WANG Huan, LI Shao-lei, SHEN Jian-xin, JIANG Ling, CHEN Sui, WANG Hai-yan
    CJAP. 2020, 36 (4): 379-384.   DOI: 10.12047/j.cjap.5976.2020.081
    Abstract   PDF (1352KB) ( 304 )
    Objective: To quantitatively analyze the effects of direct and indirect stimuli on the contraction of gastrocnemius in vivo and in vitro specimen by self-programming. Methods: All specimens were divided into four groups: indirect stimuli on specimen in vivo group (n=12), direct stimuli on specimen in vivo group (n=8), indirect stimuli on specimen in vitro group (n=12), direct stimuli on specimen in vitro group (n=8). Indirect stimuli (via sciatic nerve) and direct stimuli (acupuncture needle piercing into gastrocnemius) (stimuli starting from 0 V, cycle 3 s, increment 0.02 V, 150 times) were acted on in vivo and in vitro sciatic nerve gastrocnemius muscle specimen respectively. The effects of electric intensity on the contraction of gastrocnemius were recorded by the experimental system of BL-420F. The data were processed and analyzed by the help of self-programming, to quantitatively obtain key parameters for a single contraction. Results: ① For in vivo specimen, compared with direct stimuli, effects of indirect stimuli were as follows: the threshold intensity, half-intensity and maximal intensity of the specimen were smaller (P<0.05); the amplitude was larger, the contraction period was longer, and the rising slope was smaller (P<0.05). ②For in vitro specimen, compared with direct stimuli, effects of indirect stimuli were as follows: the threshold intensity, half-intensity and maximal intensity of indirect stimuli were smaller (P<0.05); the amplitude was larger, the contraction period was longer, and the rising slope was smaller (P<0.05). ③Compared with in vitro specimen, there was no significant difference among all the above parameters of in vivo specimen, with either direct or indirect stimuli (P>0.05). Conclusion: There is no significant difference in the features of single contraction between in vivo and in vitro specimen with either direct or indirect stimuli. However, indirect stimuli can trigger gastrocnemius to produce single contraction more easily than direct stimuli, and the amplitude is larger than that of direct stimuli.
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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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