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  • Table of Content
      28 September 2017, Volume 33 Issue 5 Previous Issue    Next Issue
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    Effects of dihydroartemisinin on radiosensitivity of Raji cells
    ZHANG Lei, CHENG Long-qiu, ZHOU Zhao, Lv Lin-lin, LIU Ge-xiu
    CJAP. 2017, 33 (5): 385-390.   DOI: 10.12047/j.cjap.5565.2017.093
    Abstract   PDF (2415KB) ( 83 )
    Objective: To study the effects of dihydroartemisinin (DHA) on radiation sensitivity of Raji cells, and explore its mechanisms.Methods: CCK8 was used to determine the effect of DHA on cell viability of Raji cells; apoptosis, intracellular reactive oxygen speies(ROS) and mitochondrial membrane potential of Raji cells were detected by flow cytometry; and the protein expressions of protein kinase B(AKT), phospho-rylated-protein kinase B(p-AKT), Bcl-2 and Bax were determined by Western blot.Results: The cells were randomly divided into four groups:control group, DHA(5μmol/L DHA), irradiation(IR, 4 Gy), IR+DHA group (4 Gy IR+5 μmol/L DHA). Compared with the other three groups, cells in DHA+IR group exhibited lower mitochondrial membrane potential (P<0.01). While the intracellular ROS content and apoptosis rate of Raji cells in DHA+IR group were increased significantly(P<0.01). In addition, compared with the other three groups, there was no significant difference in the expression of AKT, but the phosphorylation of AKT protein were significantly inhibited and the expression of Bcl-2 protein was markedly decreased. However, the expressions of Bax and Cleaved-Caspase-3 protein were markedly increased.Conclusion: DHA might activate the mitochondrial apoptotic signal via inhibiting phosphoinositide 3-kinase (PI3K/AKT) pathway and increase oxidative stress to enhance the radiosensitivity of Raji cells.
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    The role of JAK2/STAT3 signaling pathway in exercise preconditioning against myocardial apoptosis
    SUN Xiao-juan, FENG Wu-long, HOU Na
    CJAP. 2017, 33 (5): 393-397.   DOI: 10.12047/j.cjap.5473.2017.095
    Abstract   PDF (1485KB) ( 113 )
    Objective: To study the effects of Janus kinase 2/signal transducers and activators of transcription 3 (JAK2/STAT3) in exercise preconditioning (EP) against myocardial apoptosis.Methods: Eighty healthy male SD rats were randomly divided into control group(C), exercise exhaust(EE) group, EP group, and EP+JAK2 inhibitor AG490(AG) group(n=20). By using 3 days intermittent treadmill exercise, the EP animal model was established, and myocardial injury was induced by exhaustive exercise on treadmill. The changes of myocardial apoptosis were evaluated by TUNEL. The expressions of Caspase-3, p-JAK2 and p-STAT3 in heart were detected by Western blot or immunohistochemistry.Results: Compared with group C, myocardial apoptosis, and the expressions of Caspase-3, p-JAK2, and p-STAT3 in heart were increased significantly in group EE. Compared with group EE, myocardial apoptosis and the expression of Caspase-3 were decreased significantly, while the expressions of p-JAK2 and p-STAT3 were increased significantly in group EP. Compared with group EP, myocardial apoptosis and the expression of Caspase-3 were increased significantly, while the expressions of p-JAK2 and p-STAT3 were decreased significantly in group EP+AG.Conclusion: EP could increase the expressions of p-JAK2 and p-STAT3 and decrease the expression of Caspase-3 in heart, which further mitigate myocardial apoptosis. Hence, JAK2/STAT3 pathway may participate in EP against myocardial apoptosis.
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    Effects of chronic stress depression on the circadian rhythm of peripheral neuroendocrine hormone of rats
    ZHAO Yan, FU Yu
    CJAP. 2017, 33 (5): 398-402.   DOI: 10.12047/j.cjap.5529.2017.096
    Abstract   PDF (1056KB) ( 190 )
    Objective: To investigate the circadian rhythmic alterations of plasma hormone and peptide induced by chronic unpredictable mild stress(CUMS).Methods: Sixty male SD rats were randomly divided into two groups, model group and control group(n=30). Rats in model group received CUMS procedure (restraint, rotation, cage tilt, wet sawdust, cold, crowding, food or water deprivation, clamp tail and reverse light/dark cycle). Rats were randomly exposed to two stressors every day for 3 weeks. Exploratory locomotor activity was assessed by open field test and anxiety-like behavior was measured by elevated plus-maze test, and lack of pleasure by sucrose preference test. Blood samples were collected at each of six time points zeitgeber time (ZT1, 5, 9, 13, 17 and 21), 5 rats were executed at each time point. Plasma concentrations of corticosterone(CORT), melatonin (MT) and vasoactive intestinal peptide (VIP) were detected by ELISA kits, and plasma concentration of adrenocorticotropic hormone (ACTH) was determined by radioimmunoassay. The circadian rhythm changes of serum CORT, MT, VIP and ACTH in each group were compared by using cosinor.Results: Compared with control group, locomotor activity, rearing, weight gain and sucrose consumption in model group were decreased significantly. The values of the percentage of open-arm time (OT) and open arm entries (OE) were obviously lower in the model group than those in the control group. The circadian rhythm of plasma ACTH and CORT showed an obvious disorder, including phase advance and decrease in amplitude, and markedly blunted circadian rhythm. There also showed a markedly blunted circadian rhythm and decreased levels of plasma MT. VIP still had 24-hour rhythm, but the amplitude was significantly lower, peak phase also delayed for six hours, and expression was significantly higher.Conclusion: CUMS induces these peptides and hormones desynchronized from suprachiasmatic nucleus(SCN), manifested as the disturbed circadian rhythms and the abnormal hormone secretion of these synchronizers.
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    Effects of adiponectin on the anxiety and memory impairment in triple transgenic Alzheimer's disease model mice
    QU Xue-song, SHI Hui, CAO Xiu-li, DONG Xue-fan, LI Tian, JIAO Juan-juan, QI Jin-shun, WU Mei-na
    CJAP. 2017, 33 (5): 405-409.   DOI: 10.12047/j.cjap.5498.2017.098
    Abstract   PDF (920KB) ( 125 )
    Objective: To investigate the effects of adiponectin (APN) on anxiety and memory impairment of 9-month-old triple transgenic Alzheimer's disease (3xTg-AD) model mice.Methods: The 9-month-old 3xTg-AD mice and C57BL/6J mice were randomly divided into four groups (n=8 for each group):Wild type(WT)+Saline, 3xTg-AD +Saline, WT+APN and 3xTg-AD +APN group. All mice were implanted cannula in lateral ventricle and each mouse was intracerebroventricular injected with adiponectin or saline under free moving condition after 7 days recovery. The anxiety and memory ability of each mouse were observed by using open field test, object recognition task and Y-maze test.Results: ①In the open field test, compared to WT+Saline group, the time of 3xTg-AD +Saline mice spent in center was significantly decreased, and the time spent in periphery was obviously increased. However, APN treatment effectively reversed the phenomenon appeared in 3xTg-AD mice, indicating that APN could alleviate the anxiety observed in 3xTg-AD mice. ②In novel object recognition task, the discrimination index of 3xTg-AD+Saline group was (-16.7±10.1)%, significantly lower than (18.0±8.2)% in WT+Saline group (P<0.01) and (15.7±8.8)% in 3xTg-AD+APN group (P<0.01), which indicated that APN could effectively prevent the recognition memory impairment in 3xTg mice. ③In Y-maze test, the spontaneous alternation rate of 3xTg-AD +Saline group was (40.0±1.7)%,significantly lower than (56.6±4.6)% in WT+Saline group and (53.9±5.6)% in 3xTg-AD +APN group (P<0.01), which indicated that APN could prevent working memory impairment in 3xTg-AD mice.Conclusion: Adiponectin could effectively alleviate the anxiety and reverse the impairment of recognition memory and working memory of 9-month-old 3xTg-AD mice, and might play an important role in the prevention and treatment of AD.
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    Effects of the fermentation products of endophytes from Glycyrrhiza uralensis on inflammation-associated factors and Aquaporin in rat models with phlegm blocking in lung
    YANG Zhi-jun, DENG Yi, MAN Qiong, WANG Yong, YANG Xiu-juan, ZHAO Ni, LIU Liang
    CJAP. 2017, 33 (5): 410-414.   DOI: 10.12047/j.cjap.5495.2017.099
    Abstract   PDF (1841KB) ( 76 )
    Objective: To filtrate the effective fermentation products of endophytes from Glycyrrhiza uralensis by comparing expectorant effects.Methods: Ninety Wistar rats with half males and half females were randomly divided into 9 groups(n=10):including normal control group, model group, positive control group, Glycyrrhiza decoction group, 5 groups of the fermentation products of endophytes(JTZB005,JTZB006,JTZB043,JTZB060,JTZB063). The model of phlegm blocking in lung was induced by inhaling SO2 for 30 minutes and cold wind for 10 minutes twice per day for ten days. Ten days later, 10 ml/kg normal saline was administrated in normal control group and model group, 0.08 mg/kg Fufang Beimu Lvhuaan tablets was administrated in positive control group, 0.95 g/kg Glycyrrhiza water decoction and the fermentation products of endophytes were respectively administrated in Glycyrrhiza decoction group and 5 groups of the fermentation products of endophytes. All groups were given the corresponding dose by intragastric administration once a day for seven days. The changes of rats' general activities were observed during the experimental time. Two hours after the last gavage, rats were sacrificed. The upper lobe of the right lung was removed for HE and immunohistochemical staining. Pathological of lung tissues and the expressions of aquaporin-1(AQP1)and aquaporin-5(AQP5) in lung tissue were measured. In addition, the lung tissue of middle lobe of right lung were accurately weighed, and 10% homogenate were made. The supernatant was removed by centrifugation at 3 000 r/min for ten minutes. The levels of nitric oxide (NO) was determined by nitric acid reductase method. The content of tumor necrosis factor(TNF-α) and intercellular adhesion molecule 1(ICAM-1) and the concentration of cyclooxygenase-2(COX-2) in lung tissue were measured by enzyme-linked immunosorbent(ELISA) method.Results: After modeling,the levels of NO, TNF-α, ICAM-1, and the concentration of COX-2 in lung tissue were increased significantly, the expressions of AQP1 and AQP5 were decreased significantly (P<0.01). Compared with the model group, the levels of NO,TNF-α,ICAM-1 and the concentration of COX-2 in lung tissue were decreased significantly, and the expressions of AQP1 and AQP5 were increased significantly in Glycyrrhiza decoction group, JTZB005,JTZB006 and JTZB063 group(P<0.05, P<0.01).Conclusion: By filtrating, JTZB005、JTZB006、JTZB063 have expectorant effects.
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    Effects of Dexmedetomidine on the levels of proinflammatory mediators IL-1βand TNF-α in pulmonary ischemia/reperfusion injury rats and the mechanisms
    XIANG Bing-qian, HE Jin-bo, GAO Hui, LUO Zi-yin, DAI Yong-yue, WANG Wan-tie
    CJAP. 2017, 33 (5): 415-419.   DOI: 10.12047/j.cjap.5526.2017.100
    Abstract   PDF (1219KB) ( 57 )
    Objective: To evaluate the effects and mechanism of the Dexmedetomidine on the levels of proinflammatory mediators interleukin 1 beta (IL-1β) and tumor necrosis factor-α(TNF-α) in ischemia/reperfusion(I/R)rats.Methods: Fifty healthy SPF male SD rats, 250~310 g,8~12 weeks,were randomly divided into five groups(n=10):sham operation group(sham group),I/R group, dexmedetomidine group(Dex group), atipamezole group(Atip group), dexmedetomidine plus atipamezole(Dex+Atip group). The I/R model was established by clipping hilus of left lung for 30 min and then reperfusion for 2 h. Dex group, Atip group and Dex + Atip group were performed by intraperitoneal injection dexmedetomidine(20 μg/kg),atipamezole(250 μg/kg),Dexmedetomidine(20 μg/kg)+atipamezole(250 μg/kg)respectively 30 min in advance before hilus of left lung was clipped, the rest of the process was the same with I/R group. After the experiment the rats were killed and the left lung tissues to determine the lung wet/dry weight(W/D) and total lung water content(TLW); Ultra structure of lung tissues were observed under light microscope and electron microscope; IL-1β and TNF-α levels were determined by using ELISA.Results: Compared with the sham group, the W/D、TLW、IL-1β and TNF-α in other groups were increased significantly (P<0.05). The structure damages of lung tissues observed under light microscope and electron microscope in other groups were more serious than that of sham group. Compared with I/R、Atip、Dex+Atip group, the levels of W/D、TLW,IL-1β and TNF-α in Dex group were lower (P<0.05), the structure damages of lung tissues observed under light microscopy and electron microscope in Dex group were slighter. There was no significant difference of the above parameters among I/R、Atip、Dex+Atip group.Conclusion: Dexmedetomidine can alleviate ischemia/reperfusion injury in rat lung through lowering the level of proinflammatory mediators IL-1β and TNF-α,the possible mechanism may be through stimulation of α2 adrenaline receptors.
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    Effects of Dandelion polysaccharide on IL-6/STAT3 signaling pathway in ulcerative colitis rats
    WANG Qian, BIE Yu-long, WANG Dou, FAN Wen-tao
    CJAP. 2017, 33 (5): 422-425.   DOI: 10.12047/j.cjap.5525.2017.102
    Abstract   PDF (1318KB) ( 182 )
    Objective: To investigate the effects of dandelion polysaccharide on IL-6/STAT3 signaling pathway in rats with ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups (n=10):control group, model group, positive control group and dandelion polysaccharide group. The ulcerative colitis model was established by treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS). The positive control group was treated with mesalazine 10 mg/kg·d and dandelion polysaccharide group was treated with dandelion polysaccharide 10 mg/kg·d. The levels of interleukin -6(IL-6), colonic myeloperoxidase (MPO) and interleukin-6 receptor (sIL-6Rα) were measured after 4 weeks of treatment. The pathological changes of colonic mucosa were observed in rats. The gene expressions of glycoprotein 130 (gp130), transcriptional activator3(STAT3) and IL-6 were detected.Results: Compared with the normal control group, the level of serum IL-6 in the model group was significantly higher than that in the control group (P<0.01). Compared with the model group, the serum levels of IL-6 in the dandelion polysaccharide group and the methacetin group were significantly decreased (P<0.01). Compared with the model group, the MPO positive density of the rats in the dandelion polysaccharide group and the methacetin group was significantly lower than that in the normal group (P<0.01). Compared with the model group, the levels of sIL-6Rα and gp130 in the rats were significantly lower than those in the model group (P<0.01). Compared with the model group, the expressions of STAT3 and IL-6 mRNA in the intestinal tissue of the rats in the dandelion polysaccharide group and the methacetin group were decreased significantly.Conclusion: Dandelion polysaccharide can decrease the level of IL-6 in rats with ulcerative colitis, regulate the expression of sIL-6Rα and gp130 protein in IL-6/STAT3 pathway, and then down-regulate the expressions of STAT3 and IL-6 mRNA in intestinal tissue of rats, alleviate the colon inflammation state, protect and repair the mucosal tissue. Dandelion polysaccharide plays a role in the treatment of ulcerative colitis.
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    Effects of aerobic exercise and dieting on chemerin and its receptor CMKLR1 in the livers of type 2 diabetic rats
    LIN Xiao-jing, YANG Hong-fang, WANG Xiao-hui
    CJAP. 2017, 33 (5): 426-430.   DOI: 10.12047/j.cjap.5495.2017.103
    Abstract   PDF (1417KB) ( 96 )
    Objective: To investigate the effects of aerobic exercise and dieting on chemerin and its receptor chemokine like receptor-1 (CMKLR1) in the livers of type 2 diabetes mellitus (DM) rats and its possible roles on the improvements of blood glucose and lipid metabolism.Methods: Fifty male SD rats were randomly divided into control group (Con, n=6) fed with normal diet and DM model group (n=44) fed with high fat diet. After 8-week feeding, DM model rats were intraperitoneal injected streptozocin (STZ) at the dose of 30 mg/kg body weight to establish DM rats. The successfully established diabetes rats were randomly divided into 4 groups (n=6):diabetes control group (DM), diabetes exercise group (EDM), diabetes dieting group by feeding normal diet (NDM) and diabetes exercise plus dieting group (ENDM). During the following 4 weeks, exercise rats participated in moderate intensity aerobic exercise on a treadmill with a progressing increasing load for 4 weeks. Fasting blood glucose (FBG) was determined by Roche glucometer. Serum levels of triglyceride (TG), total cholesterol (TC) and low-density lipoprotein (LDL)) were detected by using full automatic biochemical analyzer. The mRNA and protein expressions of chemerin and CMKLR1 in the livers were detected by real time PCR and Western blot, respectively.Results: Accompanied with the increased serum levels of TC, TG, LDL and FBG in DM rats, the mRNA and protein levels of chemerinand CMKLR1 in the livers of DM rats were significant up-regulated. The serum levels of TC, TG, LDL and FBG were reduced in EDM, NDM and ENDM rats compared with those of DM rats, meanwhile,decreased protein levels of chemerin were found in the livers of EDM, NDM and ENDM rats with most significant in ENDM rats while increased protein levels of CMKLR1 in the livers of NDM and ENDM rats.Conclusion: Four weeks of aerobic exercise with or without dieting decreased chemerin and increased CMKLR1 in the livers of DM rats at the protein levels, which might be attributed to the improvement of blood glucose and lipid metabolism of DM rats.
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    Effects of yellow wine polyphenols on cardiomyocyte apoptosis in diabetic cardiomyopathy rats
    PAN Sun-lei, LIN Hui, LUO Hang-qi, GAO Fei-dan, MENG Li-ping, GUO Yan, GUO Hang-yuan, CHI Ju-fang
    CJAP. 2017, 33 (5): 431-435.   DOI: 10.12047/j.cjap.5553.2017.104
    Abstract   PDF (1103KB) ( 91 )
    Objective: To investigate the effects of yellow wine polyphenols on the apoptosis of cardiomyocytes in diabetic cardiomyopathy rats.Methods: Thirty SD rats were randomly divided into control group (Control), diabetic cardiomyopathy group (DCM) and diabetic cardiomyopathy treated with yellow wine polyphenols group (DCM+YWP). A single intraperitoneal injection of 65 mg/kg streptozotocin (STZ) was utilized to establish a rat model of DCM. The rats in control group were treated with citrate buffer at the same dose of a single intraperitoneal injection. DCM+YWP group were treated with 18 mg/kg Yellow wine polyphenols by ig after modeling. After treated for 12 weeks, the general condition of rats were observed. The cardiac structure and function of the rats were observed by Doppler echocardiography. The ultrastructure of myocardium were observed using electron microscopy. The inflammation index of myocardial tissue was detected by enzyme-linked immunosorbent assay (ELISA). The oxidative stress in myocardial tissues was assessed by oxidative stress detection kits. The expressions of Bax, Bcl-2 and Caspase-3 (cleaved) in myocardial were detected by Western blot.Results: Compared with DCM group, the blood glucose levels and body weight of rats in the DCM+YWP group were not changed significantly. Echocardiography showed that left ventricular end-diastolic diameter, left ventricular end-systolic diameter were decreased (P<0.05), while fractional shortening and E/A ratio and Ea/Aa ratio were elevated (P<0.05). The levels of tumor factor-α(TNF-α), interleukin 1β(IL-1β) and interleukin 6(IL-6) in myocardium were decreased (P<0.05). The levels of oxidative stress malondiadehyde(MDA) were decreased and Superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) were increased in myocardial tissue (P<0.05). The expression levels of Bax and Caspase-3 (cleaved) protein in myocardium were decreased (P<0.05), and the expression of Bcl-2 protein was increased (P<0.05).Conclusion: Yellow wine polyphenols can improve the diabetic cardiomyopathy rat cardiac function, attenuates inflammation and oxidative stress in diabetic rats, inhibit the apoptosis of cardiomyocytes in diabetic cardiomyopathy.
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    Effects of weight-bearing ladder and aerobic treadmill exercise on learning and memory ability of diabetic rats and its mechanism
    TANG Liang, KANG Yi-ting, YIN Bo, SUN Li-jun, FAN Xiu-shan
    CJAP. 2017, 33 (5): 436-440.   DOI: 10.12047/j.cjap.5570.2017.105
    Abstract   PDF (1161KB) ( 101 )
    Objective: To investigate the effects of climb ladder and aerobic treadmill exercise on learning memory ability in diabetic rats and explore its possible mechanisms.Methods: Forty male rats were randomly divided into normal control group (NC), diabetic control group (DC), diabetic loading ladder group (DL) and diabetic aerobic treadmill group (DA), diabetes was induced by a single intraperitoneal injection of STZ. In the evening, the DL group were trained three cycle (10 times/cycle) with weight-bearing climbing ladder, 2 min intervals, 6 days/week, lasted for six weeks. The DA group was trained on a motor-driven treadmill at a speed of 20 m/min (0 incline), 30 min/day, 6 days/week, lasted for six weeks. Morris water maze was used to detect the learning and memory ability of rats after modeling success and after exercise intervention. After the last water maze test, the rats were killed to obtain the hippocampus. RT-QPCR was used to detect the gene expressions of brain derived neurophic factor (BDNF), tropomyosin receptor kinase B (TrkB) and cAMP-response element binding protein (CREB).Results: Compared with NC group, the expressions of BDNF and CREB gene in hippocampus of DC group and the learning and memory ability were significantly decreased. Compared with DC group, the expression of BDNF and CREB in hippocampus of DL and DA rats was significantly up-regulated and the learning ability was significantly increased. The TrkB gene of hippocampus in DL rats was significantly up-regulated and the spatial memory ability was significantly improved. Compared with the DA group, the TRKB and CREB genes in the hippocampus of DL group were significantly up-regulated.Conclusion: Aerobic treadmill exercise and weight-bearing ladder exercise have a positive effect on the learning ability of diabetic rats, while the weight-bearing ladder exercise improves the memory ability of diabetic rats better than aerobic exercise. These effects may be related to the up-regulation of BDNF/TrkB/CREB signaling pathway.
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    Effects of curcumin on the recovery of hind limb function after spinal cord injury in rats and its mechamism
    HAO Qin, WANG Hui-wei, YU Qian, SHEN Juan, ZHAO Lin, SHI Fang-fang, CHEN Mei-mi, YANG Yan-ling
    CJAP. 2017, 33 (5): 441-444.   DOI: 10.12047/j.cjap.5548.2017.106
    Abstract   PDF (1199KB) ( 123 )
    Objective: To investigate the effects of curcumin on motor function, and to explore the neuroprotective mechanism of curcumin after the spinal cord injury in rats. The study will theoretical and experimental evidence for curcumin's clinical treatment.Methods: HI-0400 spinal cord impactor was used to prepare animal models of acute of spinal cord injury. One hundred and five clean and healthy rats were randomly divide into three groups:sham operation group (Sham) spinal cord injury group (SCI) and curcumin group (SCI+CUR). Intragastric administration was administrated after 30min of the spinal cord injury model, after 1 time a day, until the death. SCI+CUR group was intragastric administration with curcumin (100 mg/kg) of 0.5% carboxymethyl cellulose sodium, and Sham and SCI group were treated with the same dose of 0.5% carboxymethyl cellulose sodium. The motor function recovery of 3,7,14,21 and 28 days after spinal cord injury were evaluated by basso,beatlie,bresnahan (BBB) score. The spinal cord tissue and blood samples were collected at postoperative 12 h, 1 d, 3 d and 7 d respectively, NF-kappa B was detected by immunofluorescence, Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry. The expression of Bcl-2 and Bax was detected by Elisa.Results: The statistic difference of BBB score between SCI group and CUR group in 3 day was not statistically significant. It was found that the 7,14,21 and 28 days BBB score in CUR group were statistically significant higher than that in SCI group(P<0.05).The expression of inflammatory factor NF-kappa B appeared in 12 h after spinal cord injury, 1 day peaked and 3 days decreased. In SCI+CUR group, the expression of NF-kappa B at each time point was similar to the SCI group, and there was a difference between group SCI+CUR and SCI group in 1day.There was no obvious staining of Bax and Bcl-2 in Sham group. The staining of Caspase-3 and Bax in SCI+CUR group was significantly weaker than that in SCI group, while Bcl-2 was stronger.Conclusion: Curcumin can promote the recovery of hindlimb motor function after spinal cord injury in rats.The mechanism is through inhibition of NF-K B to prevent inflammation; And inhibition the expression of Bax and Caspase-3, and promotion the expression of Bcl-2 to prevent apoptosis, so as to accelerate the recovery of motor function in the rats after spinal cord injury.
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    Establishment of neuritis migration model induced by Slit 2N and silk fibroin mixture in the rat hippocampal neurons in vitro
    GANG Lin, LI Yi-peng, LU Lei, YANG Kai, SUN Zhong-lei, CHEN Xu-yi, TU Yue
    CJAP. 2017, 33 (5): 445-449.   DOI: 10.12047/j.cjap.5563.2017.107
    Abstract   PDF (1427KB) ( 54 )
    Objective: To explore a simply feasible and affordable method to establish neuritis migration model induced by Slit 2N and silk fibroin mixture in the rat hippocampal neurons in vitro.Methods: Neurons were derived from SD rat hippocampal tissues and cultured with a special cell culture-plate in vitro. The cultured neurons were divided into four groups, named as control group, pure silk fibroin, pure Slit 2N and Slit 2N mixture with silk fibroin (mixture group), and 50 different neurons were randomly selected in each group. Moreover, we photographed and recorded the soma coordinate and added the silk fibroin, Slit 2N and mixture to each neurite with a distance of 100 μm, except control group. Record again after 30 min. Property and positive rate of cells were identified by immunofluorescence staining.Results: Neurites of the pure Slit 2N group and the mixture group became shorter, and there was no significant change in the pure silk fibroin and control groups. The results showed that the average duration and length difference before and after changed were in descending order of mixture group, Slit 2N group, silk fibroin group (P<0.05), and the silk fibroin and control groups were no significant change (P>0.05). The positive rate of MAP-2 in four groups was more than 90%.Conclusion: There were no significant effects of Silk fibroin on induced by Slit 2N in rat hippocampal neuron migration. It had an effect on reducing Slit 2N diffusion rate and extending its working time. It provides an advantageous construction method in vitro on based 3D directed neural repair for the treatment of central nervous system diseases.
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    Effects of triptolide on the apoptosis and H3K4 protein methylation in multiple myeloma cells
    XU Cheng-bo, LIAO Bin, SHEN Jian-zhen, FU Hai-ying, LIN Ting
    CJAP. 2017, 33 (5): 450-455.   DOI: 10.12047/j.cjap.5483.2017.108
    Abstract   PDF (0KB) ( 243 )
    Objective: To investigate the effects of triptolide (Chinese Traditional Medicine)on the apoptosis and H3K4 protein methylation in multiple myeloma cells.Methods: The RPMI8226 cells were cultured with different concentrations(10,20,40,80 and 160 nmol/L)of triptolide for different incubation time (24 h,48 h and 72 h). The inhibition of triptolide on RPMI8226 cell proliferation was detected by MTT assay. Apoptosis and cell cycle distribution were evaluated by flow cytometry.The expressions of H3K4me2 and trimethylation of histone H3 lysine 4(H3K4me3) in RPMI8226 cells were assayed by Western blot. The changes of expressions of histone methylase SET and MYND domain containing 3(SMYD3) and histone demethylase lysine specific demethylase 1(LSD1) in RPMI8226 cells were verified by qRT-PCR.Results: Triptolide had obvious inhibitive effects on proliferation of RPMI8226 cells and showed a dose-and time-dependent manner(P<0.05). Triptolide induced apoptosis and G2/M cell cycle arrest in a dose-dependent manner(P<0.05). Triptolide decreased histone H3K4me2 and H3K4me3 expression in a dose-dependent manner(P<0.05, P<0.01). SMYD3 was significantly depressed at protein expression in a dose-dependent manner(P<0.05), but LSD1 was up-regulated (P<0.05).Conclusion: Triptolide could inhibit RPMI8226 cell proliferation,induce the apoptosis and cause G2/M arrest,meanwhile,significantly inhibit the protein expressions of H3K4me2 and H3K4me2 with alter the expression of SMYD3 and LSD1.The effects is probably related to the antitumor mechanism of MM cells induced by triptolide.
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    SR 59230A on the expression of MicroRNAs in myocardium of heart failure rats
    JING Jia-ni, LI Fan-lu, WANG Xi, WAN Xin, ZHAO Qian-qian, CUI Xiang-li
    CJAP. 2017, 33 (5): 456-460.   DOI: 10.12047/j.cjap.5561.2017.109
    Abstract   PDF (1206KB) ( 93 )
    Objective: To investigate the effects of β3-adrenoceptors(β3-AR) inhibitor SR 59230A on MicroRNAs expression in rat myocardium with chronic heart failure and the related mechanisms.Methods: One hundred male SD rats were randomly divided into sham operated group(40)and chronic heart failure(CHF)group(60). Coronary artery ligation was used to induce CHF. Then the rats in CHF group were further randomly divided into CHF control group and CHF+SR 59230A group (CHF+SR). Rats in the sham group were divided into sham control group and sham+SR 59230A group (Sham+SR). The rats in Sham+SR group and CHF+SR group were treated with 1 ml SR 59230A(85 mmoL/L in 0.9% saline)twice a day for seven weeks by intraperitoneal injection, while the rats in control groups were injected with the same amount of saline for seven weeks separately. miScript miRNA PCR Arrays were used to determine the expression profile of MicroRNAs. Immunohistochemistry was used to evaluate the distribution of the related proteins in the heart tissue sections. Western blot was used to detect the expressions of nuclear factor-kappaB(NF-κB),p53 and p53-Phospho-Serine 15 in the heart.Results: ①After in vivo blockade of β3-AR by SR 59230A, there were 18MicroRNAs down-regulated in sham control group and CHF control group. Within them, 6 MicroRNAs were related to NF-κB signaling pathway, they were miR-125b-5p,miR-143-3p,miR-145-5p,miR-26a-5p,miR-30a-5p and miR-320-5p. ②Slides from the heart tissue showed that NF-κB was distributed both in nucleus and cytoplasm, while p53 in cytoplasm was more than that in nucleus in heart tissue sections. The expressions of NF-κB and p53 were higher in the CHF control group than those in the sham control group(P<0.05), but were lower in CHF+SR group than those in CHF control group(P<0.05),while they were elevated in Sham+SR group compared to the sham control group(P<0.05). ③ Compared with the sham control group, the protein expression of NF-κB p65 was increased significantly in the CHF control group (P<0.05). After treated with SR59230A in vivo,the protein expressions of NF-κB and p53-Phospho-Serine 15 were decreased significantly in CHF rats(P<0.05),while the protein expressions of NF-κB, p53 and p53-Phospho-Serine 15 proteins were increased in the sham rats (P<0.05).Conclusion: Blocking of β3-AR improved the damaged heart in CHF rats; β3-AR caused the change of MicroRNAs expression, and it related to NF-κB signal pathway.
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    Effects of different intensities exercise combined with resveratrol on RBP4 in aged obese rats
    CUI Jia-qing, BAI You-ping, SU Min, SONG Wei-wei, LIN Chan, WU Lin, ZHANG Hao-yue
    CJAP. 2017, 33 (5): 461-465.   DOI: 10.12047/j.cjap.5527.2017.110
    Abstract   PDF (958KB) ( 104 )
    Objective: To investigate the effects of different intensity exercise combined with resveratrol on retinol binding protein 4(RBP4) mRNA and protein expression in visceral adipose tissue and plasma RBP4 concentration of aged obese rats.Methods: Eighty male SD rats aged 3 weeks were randomly divided into the control group and the experimental group. Rats of the control group(C) were fed with ordinary feed of 6% fat (n=12). The experimental group consisted of 68 rats, which were fed with 36%~40% high fat feed in three stages. The rat model of elderly obesity was established successfully. Twenty-four obese rats were selected and randomly divided into 4 groups, including obese group (CO), resveratrol obese group (RO), low intensity exercise combined with resveratrol group (LRO) and moderate intensity exercise combined with resveratrol group (MRO). There were 6 rats in each group. The exercise intensities of the LRO group and MRO group were (12 m/min×15 min) and (15 m/min×15 min). Every day was exercised for 60 minutes. The rats in RO, LRO or MRO were treated with resveratrol at the dose of 52.5 mg/kg·d, while the rats in the control group were fed with the same amount of purified water, continued intervention for 8 weeks. After 8 weeks, samples of blood, adipose tissue of around the kidney, testicle, vessel and visceral were collected. Blood glucose and plasma RBP4 concentration were measured. Insulin sensitivity(ISI) was calculated. The expressions of RBP4 mRNA and protein were also determined.Results: Compared with normal (C) group, the expressions of RBP4 mRNA and protein and plasma concentration and plasma glucose of the model (CO) group were increased obviously (P<0.05, P<0.01). ISI was decreased obviously (P<0.05). Compared with model (CO) group, the expressions of RBP4 mRNA and protein and plasma concentration and plasma glucose of the RO and LRO and MRO groups were decreased significantly(P<0.05, P<0.01), while ISI was increased markedly (P<0.05). Compared with the RO and LRO groups, the expressions of RBP4 mRNA and protein and plasma concentration and plasma glucose of the MRO group were decreased, while ISI was increased, but there was no significant difference.Conclusion: Different intensities exercise combined with resveratrol could reduce the RBP4 mRNA and protein expression in visceral adipose tissue and plasma RBP4 concentrations of aged obese rats, but less affected by exercise intensity.
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    Puerarin induces apoptosis in A549 cells
    JIN Xin, SUN Pan-pan, HONG Yun, YU Le, LI Shu
    CJAP. 2017, 33 (5): 466-469.   DOI: 10.12047/j.cjap.5535.2017.111
    Abstract   PDF (1138KB) ( 255 )
    Objective: To explore the effect of puerarin induced apoptosis in human non-small cell lung canner A549 cells and its possible mechanisms.Methods: A549 cells were cultivated in vitro. When 80%~90% confluence was reached, they were treated with puerarin in different concentration (60、120、240 μg/ml). The effects on cell apoptpsis were examined by CCK-8 colorimetry. Characteristic morphological changes of apoptosis were observed by AO/EB staining. The apoptosis of A549 cells were detected by flow cytometry analysis. The proteins level of Apelin and APJ were tested by Western blot.Results: CCK-8 test showed that puerarin could inhibit the proliferation of A549 cells in a time and concentration dependent manner. Flow cytometry analysis indicated that puerarin could make A549 cells apoptosis. Compared with the A549 cells group, the protein expression of Apelin/APJ was decreased in puerarin groups.Conclusion: Puerarin induces apoptosis in A549 cells, and its mechanism may through the regulation of Apelin/APJ expression.
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    Effects of TAK gene silencing on the expressions of IL-6 and IL-8 induced by TNF-α in fibroblast-like synoviocytes
    MO Xuan-rong, XIE Jiang-wen, LV Guo-ju, KE Yu-ping, LUO Xin-jing
    CJAP. 2017, 33 (5): 471-475.   DOI: 10.12047/j.cjap.5494.2017.113
    Abstract   PDF (1548KB) ( 88 )
    Objective: To investigate the effects of silencing transforming growth factor-β activating kinase 1 (TAK1)on the expressions of IL-6 and IL-8 induced by TNF-α in fibroblast-like synoviocytes, and to explore the role of TAK1 in rheumatoid arthritis (RA).Methods: The synthesized TAK1 siRNA and scrambled siRNA (ScRNA) were transferred into cultured RA fibroblast-like synoviocyte line MH7A by lipofectamine. The expressions of the pro-inflammatory mediator IL-6 and IL-8 and the levels of phospho-P38(p-P38), phospho-C-Jun NH2-terminal kinase(p-JNK), phospho-extracellular signal-regulated kinase(p-ERK), phospho-p65(p-p65) and nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha(IκBa) were examined.Results: Silencing of TAK was demonstrated in synoviocytes transfected by TAK siRNA. TAK1 silencing markedly attenuated the expression of IL-6 and IL-8 in the presence of TNF-α. TAK1 silencing inhibited the activation of p38 and JNK MAPK. TAK1 silencing also inhibited activation of nuclear factor-κB (NF-κB).Conclusion: TAK1 silencing attenuated the expression of IL-6 and IL-8 in synoviocytes induced by TNF-α via inhibiting the activation of p38, JNK MAPK and NF-κB.
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    Effects and mechanisms of lotus leaf water extract on lipid metabolism of adult experimental obesity rats
    LI Mao-mao, HUANG Xin-yuan, LIANG Qiang-kun, LI Yun-xia, YAN Yuan, TIAN Zhi-feng, QIU Xiao-qing, ZHANG Xiao-wen, SONG Qing, LI Hong-fang
    CJAP. 2017, 33 (5): 476-480.   DOI: 10.12047/j.cjap.5409.2017.114
    Abstract   PDF (1432KB) ( 170 )
    Objective: To investigate the lipid metabolic effect and mechanism of water extract of lotus leaves(Traditional Chinese Medicine).Methods: Isolated SD rat lipid tissues were suspended in organ baths containing Krebs solution, and the effect of lotus leaf water extract on free fatty acids (FFA) release was observed; The experimental obesity rat model was established by feeding them high glucose and fat diets, then the changes of body weight and blood lipid were measured in the model rats after intragastric administration with water extract of lotus leaves for four weeks, and the expressions of peroxisome proliferator-activated receptor gamma (PPAR-γ) and leptin were examined by RT-PCR and immunohistochemical.Results: The ex vivo experiment showed that water extract of lotus leaves effectively promoted the FFA release from isolated lipid tissues. In vivo experiment, similarly to Orlistat, water extract of lotus leaves(60 mg/kg)markedly decreased the body weight and blood lipid of experimental obesity rats(P<0.05), and obviously reduced the expressions of PPAR-γ and leptin(P<0.05).Conclusion: Water extract of lotus leaves greatly improves the expression of PPAR-γ and leptin, which can promote the lipid mobilization and dissolution, reduce the body weight and blood lipid of adult rats with experimental obesity, therefore is expected to be developed into lipid-lowering diet pills.
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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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