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Effects of resveratrol on ulcerative colitis in mice and its mechanism
LIU Xin, WU Ya-li, LIU Kai-li, CUI Xiang-li, DU Xin-xin, ZHANG Wen-qin
CJAP    2019, 35 (5): 447-453.   DOI: 10.12047/j.cjap.5826.2019.097
Abstract123)      PDF(pc) (1689KB)(684)       Save
Objective: To investigate the anti-ulcerative colitis mechanism of resveratrol through regulation of Wnt/β-catenin signaling pathway. Methods: ①The experiment of ulcerative colitis induced by dextran sulfate sodium salt (DSS): 28 C57BL/6 mice were randomly divided into four groups including control group(n=7), DSS group(n=7), DSS+Resveratrol (DSS+Res) group(n=7) and Res group(n=7). The experiment lasted for 3 weeks. Ulcerative colitis of mice was induced by drinking DSS water and treated with resveratrol by intragastric administration. The mice were weighed daily and their activities and state of feces were recorded. After that, the mice were euthanized, the spleens were weighed, and the colonic length was measured.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the colon, and the expression of miR-31 in colonic tissue was detected by quantitative real-time PCR (qPCR). The expressions of β-catenin and Cyclin D1 were measured by Western blot. ②In vitro experiment: HCT 116 cells were treated with resveratrol at 10 mg/ml, the expressions of β-catenin, LDL receptor related protein-6 (LRP-6), frizzled-3 (FZD3) and c-Myc were detected. The expression of β-catenin was also detected in HCT 116 cells transfected with miRNA-31 mimic and miRNA-31 inhibitor. Results: ① The body weight was decreased in DSS group, the activity was decreased and blood stool appeared. The colonic length of mice was shortened, the spleen was enlarged and the tissue was damaged seriously in DSS group. While the above symptoms were improved after resveratrol treatment. ② Resveratrol inhibited the expressions of miRNA-31, β-catenin and CyclinD1 in ulcerative colitis mice, and also down-regulated the expressions of β-catenin, LRP-6, FZD3 and c-Myc in HCT 116 cells. After transfection of miRNA-31 inhibitor, the expression of β-catenin was decreased in HCT 116 cells. Conclusion: Resveratrol suppresses DSS induced colitis by down-regulation of Wnt signal pathway. The down-regulation of Wnt signal may be related to miRNA-31.
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CJAP    2018, 34 (4): 363-366.   DOI: 10.12047/j.cjap.5584.2018.083
Abstract88)      PDF(pc) (1499KB)(657)       Save
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Improvement of hydrogen on liver oxidative stress injury in chronic intermittent hypoxia rats
YANG Sheng-chang, CHEN Ling-ling, FU Tian, LI Wen-ya, JI En-sheng
CJAP    2018, 34 (1): 61-64.   DOI: 10.12047/j.cjap.5484.2018.016
Abstract108)      PDF(pc) (1069KB)(639)       Save
Objective: To explore the effects of hydrogen on liver injury in chronic intermittent hypoxia rats and the related oxidative stress mechanism.Methods: Twenty-four male adult SD rats were randomly divided into 3 groups(n=8):the normoxia group (Norm), the chronic intermittent hypoxia group (CIH), the chronic intermittent hypoxia and hydrogen group (H2 + CIH). Rats in Norm group were exposed in air, those in the other 2 groups suffered from chronic intermittent hypoxia conditions for 5 weeks. Before the CIH treatment, rats in H2+CIH group inhaled hydrogen gas at 67% concentration for 1 hour. The serum biochemical indicators of oxidative stress, pro-inflammatory cytokine, liver enzyme and blood lipid were inspected after five weeks treatment, the pathological changes of liver tissue were also observed in the transmission electron microscope.Results: Compared with Norm group, the microstructure of liver cells was severely injured, and the serum levels of glutamic-pyruvic transaminase(ALT),glutamic-oxalacetic transaminase (AST) were significantly higher in CIH group (P<0.05); the serum level of 8-hydroxy-2 deoxyguanosine(8-OHdG) and interleukin-6(IL-6) was significantly higher, the serum level of superoxide dismutase (SOD) was significantly lower. Compared with CIH group, the pathology of liver microstructure were significantly improved and the serum levels of ALT, AST were significantly lower in H2+CIH group (P<0.05); the serum levels of 8-OHdG and IL-6 were significantly lower, the serum level of SOD was significantly higher. Compared with Norm group, the serum level of IL-1 was higher, the serum level of TC, TG, and low density lipoprotein(LDL) were lower, but there was no statistical difference with those in CIH group. There was no statistical difference in the serum level of high density lipoprotein (HDL)among the three groups.Conclusion: Pre-treatment with hydrogen could improve the liver injury caused by chronic intermittent hypoxia, and reducing oxidative stress level for protecting the liver cells damage.
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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
Abstract479)      PDF(pc) (1709KB)(489)       Save
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 nitidine chloride on ulcerative colitis in mice and its mechanism
WU Ya-li, LIU Xin, LIU Kai-li, CUI Xiang-li, WANG Chun-fang
CJAP    2019, 35 (6): 525-529.   DOI: 10.12047/j.cjap.5827.2019.115
Abstract35)      PDF(pc) (2378KB)(477)       Save
Objective: To investigate the protective effects of nitidine chloride (NC) on dextran sodium sulfate (DSS) - induced ulcerative colitis (UC) in mice by targeting miR-31 and its underlying mechanisms. Methods: DSS at the concentration of 1% was used to induce UC in mice. Thirty C57BL/6 male mice were randomly divided into four groups: normal control group (n=7), DSS group (n=8), DSS + NC group (7.27 mg/kg) (n=8) and NC group (n=7). DSS was added in drinking water, and NC was administrated by gavage. The period of modeling lasted for 3 weeks. The control group and NC group drank sterile water every day, DSS group and DSS + NC group drank 1% DSS water in the first week, normal water in the second week and 1% DSS water in the third week. In the last week of modeling, mice in control group and DSS group were given 0.5% CMC-Na by gavage, while mice in DSS + NC group and NC group were given NC by gavage. After the establishment of the model, the disease activity index (DAI) related to colitis was observed, the pathological score of colon tissue was evaluated by HE staining, the expression level of miR-31 in colon tissue was detected by qPCR, and the protein expressions of NF -κ B and COX-2 in colon tissue were detected by Western blot. Results: ① Compared with DSS group, the DAI in the DSS + NC group was decreased (P<0.01). The colonic pathological injury was obviously ameliorated after treated by NC. ② Compared with normal control group, the expression of miR-31 in colonic tissue of DSS group was increased significantly(P<0.01), compared with DSS group, the expression of miR-31 was decreased after treatment with NC(P< 0.05). ③ Compared with DSS group, the levels of inflammatory protein NF-κB and COX-2 in DSS + NC group was decreased significantly (P<0.05). Conclusion: Nitidine chloride has obvious therapeutic effects on DSS induced mouse colitis, and its anti-inflammatory mechanism is related to the down-regulation of miR-31 expression.
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Reproduction,genotype identification and evaluation of APP/PS1 transgenic mice
TAN Long, LI Hai-qiang, LI Yi-bo, LIU Wei, PANG Wei, JIANG Yu-gang
CJAP    2018, 34 (2): 111-114.   DOI: 10.12047/j.cjap.5541.2018.027
Abstract768)      PDF(pc) (1106KB)(435)       Save
Objective:To identify the genotype of (APP/PS1) transgenic mice and evaluate the changing of cognitive and behavioral fu nctions, provide an effective animal model for the Alzheimer's disease (AD) research. Methods:Male APP/PS1 transgenic mice mated with female APP/PS1 transgenic mice, and the genotype of their filial mice was identified by PCR. The APP +/PS1 + mice were assigned into AD model group (AD group, n=8), and the APP/PS1 mice were assigned into control group (CT group, n=8). The Morris water maze test was carried out to detect the capacity of learning and memory of mice. After that, the mice were sacrificed and the brain tissues were sampled and stained by HE and congo red for the pathological examination. Results:①A APP/PS1 genome DNA about 360 bp size was detected. The methods of feeding and breeding were successful to attain APP/PS1 transgenic mice.②Statistical significance was found in the differences of the capacity of learning and memory between 7-month-old APP/PS1 positive mice and negative mice (P<0.05).③The results of HE stain showed that the structure and cellular morphology of hippocampus of AD mice were obviously abnormal. The results of congo red stain showed that positive amyloid plaque was observed in brains of AD mice. Conclusion:APP/PS1 transgenic mice present typical symptoms and behaviors of Alzheimer's disease. The transgenic mouse is an effective tool for the research and prevention of AD.
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Prognosis-related miRNA bioinformatics screening of lung adenocarcinoma and its clinical significance
ZHANG Hao-min, YANG Bo, CHEN Hong-fei, CHI Xiao-hua, XI Yi-bo, CHEN Xi-meng, GUO Bin, HE Pei-feng, LU Xue-chun
CJAP    2018, 34 (6): 530-535.   DOI: 10.12047/j.cjap.5755.2018.118
Abstract313)      PDF(pc) (1479KB)(434)       Save
Objective: To investigate the prognosis-related miRNA histological features and clinical significance of lung adenocarcinoma. Methods: Using The Cancer Genome Atlas (TCGA) data, the miRNA expression profile data of human lung adenocarcinoma were searched for differential analysis, and the prognosis-related miRNAs were screened by Cox risk regression model. The targeted miRNAs were predicted by mirwalk analysis platform, KEGG functional enrichment analysis, and finally, predict the function of prognosis-related miRNAs. Results: A total of 46 differential miRNAs in lung adenocarcinoma were screened, including 19 up-regulated and 27 down-regulated. Six prognostic-related miRNAs were screened by Cox survival analysis, namely hsa-mir-21, hsa-mir-142, hsa-mir-200a high expression, hsa-mir-101, hsa-let-7c, hsa-mir-378e low expression, hsa-mir-21 and hsa-mir-378e were associated with poor prognosis in patients with lung adenocarcinoma, and the survival time was shortened significantly (P<0.05, AUC=0.618). KEGG analysis showed that the above prognosis-related miRNA targeting regulatory genes were related with immune response pathways, miRNA and cancer pathways, metabolic pathways and so on. Conclusion: Hsa-mir-21 and hsa-mir-378e are associated with poor prognosis of lung adenocarcinoma, and may be used as a molecular marker for prognosis of lung adenocarcinoma after further clinical verification.
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Effects of Radix Hedysari, Radix Astragalus and compatibility of Angelica Sinensis on blood deficiency model mice induced by cyclophosphamide
YANG Xiu-juan, YANG Zhi-jun, LI Shuo, JI Xiu-jia, Ning Yan-mei, WANG Yan
CJAP    2018, 34 (6): 550-554.   DOI: 10.12047/j.cjap.5698.2018.123
Abstract149)      PDF(pc) (1246KB)(422)       Save
Objective: To study the intervention effects of Radix Hedysari, Radix Astragalus and compatibility of Angelica Sinensis on blood deficiency model mice induced by cyclophosphamide (CTX). Methods: The mice were randomly divided into 7 groups, 10 mice each group. The blood deficiency model was established by CTX. The blank group and model group were treated with saline by gavage, while mice in positive group were administered with Lvjiaobuxue granule. Four dosage group were administered with Radix Hedysari, Radix Hedysari-Radix Angelica Sinensis(5:1), Radix Astragalus and Radix Astragalus-Radix Angelica Sinensis(5:1) water decoction. All the drugs were administered to mice for consecutive 7 d. The contents of red blood cell (RBC), lymphocyte(LYM), hematocrit (HCT), white blood cell (WBC), platelet (PLT) were detected by hematology analyzer, while thymus index(TI), spleen index(SI), reticulocyte (RC), marrow karyocyte (MK) were calculated, and the femur by pathological section were observed by microscope. Results: Compared with blank group, the contents of RBC, WBC, HCT, PLT, LYM were decreased in model group (P<0.05). Compared with model group, the contents of RBC, WBC, HCT, PLT, LYM, RC and marrow karyocyte were increased in Hedysari-Angelica Sinensis(5:1) and Astragalus Angelica Sinensis(5:1) (P<0.05), at the same time, the pathological damage of femur could be improved. Conclusion: The effect of enrichment blood on blood deficiency model mice in Hedysari-Angelica Sinensis (5:1) and Astragalus-Angelica Sinensis(5:1) were superior to Hedysari and Astragalus.
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Changes of apoE protein expression in lung of mice with hypoxic pulmonary arterial hypertension
LIU Ling-yan, HUANG Tian-peng, HAO Jia-le, CHEN Ran, FAN Xiao-fang, GONG Yong-sheng, MAO Sun-zhong
CJAP    2018, 34 (6): 492-495.   DOI: 10.12047/j.cjap.5723.2018.110
Abstract225)      PDF(pc) (1123KB)(414)       Save
Objective: To observe the changes of apolipoprotein E (apoE) protein expression of pulmonary tissue in mice with pulmonary hypertension induced by hypoxia. Methods: The animal model of hypoxic pulmonary hypertension was established by exposing the mice to isobaric hypoxic chamber for 3 weeks (23 h/d, regular chow feed).Twenty male wild type (WT) C57BL/6 mice and twenty apoE gene knockout (apoE-KO) mice were randomly divided into normoxia group and hypoxia group. The plasma concentrations of low density lipoprotein (LDL), high density lipoprotein (HDL) and total cholesterol were detected by ELISA method. The protein expression of apoE in lung and liver, and peroxisome proliferators-activated receptor gamma (PPARγ) in lung were measured by Western blot.Results: ①In WT mice, the right ventricular systolic pressure (RVSP) and the weight ratio of right ventricle (RV) to left ventricle plus septum (LV+S) of hypoxia group were significantly higher than those of normoxia group by 68% and 59% (P<0.05), respectively. The plasma concentration of HDL and HDL/LDL of hypoxia group were significantly lower than those of normoxia group by 17% and 40% (P<0.05), respectively.The protein expression of apoE in lung and in liver of hypoxia group were significantly down-regulated than those of normoxia group by 48% and 52% (P<0.05), respectively.The protein expression of PPARγ in lung was significantly down-regulated than that of normoxia group by 37%(P<0.05).RVSP were significantly negative correlated with the protein levels of apoE and PPARγ in lung (P<0.01).② In apoE-KO mice, RVSP and the weight ratio of RV to LV+S of hypoxia group were significantly higher than those of normoxia group by 96% and 86% (P<0.05), respectively.RVSP and RV to (LV+S) of hypoxia group in apoE-KO mice were significantly higher than those of hypoxia group in WT mice by 29% and 24% (P<0.05), respectively.Conclusion: Down-regulated expression of apoE in lung tissue participates in the pathological proceeding of pulmonary hypertension induced by hypoxia.
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Bioinformatics analysis of genes related to poor prognosis of human hepatocellular carcinoma and its clinical significance
XI Yi-bo, ZHANG Hao-min, YANG Bo, CHEN Xi-meng, HE Pei-feng, LU Xue-chun
CJAP    2019, 35 (1): 90-96.   DOI: 10.12047/j.cjap.5764.2019.021
Abstract190)      PDF(pc) (3467KB)(410)       Save
Objective:To screen genes associated with poor prognosis of hepatocellular carcinoma (HCC) and to explore the clinical significance of these genes. Methods: The proper expression profile data of HCC was obtained from the Gene Expression Omnibus (GEO) database, and the differentially expressed genes (DEGs) were identified by differential expression analysis. The DAVID and String database were used for function enrichment analysis and to construct the protein-protein interaction (PPI) network respectively. The Cancer Genome Atlas (TCGA) database and the Cox Proportional Hazard Model were used for prognosis analysis of the DEGs. Results: A eligible human HCC data set (GSE84402) met the requirements. A total of 1141 differentially expressed genes were identified, including 720 up-regulated and 421 down-regulated genes. The results of function enrichment analysis and PPI network performed that CDK1、CDC6、CCNA2、CHEK1、CENPE 、PIK3R1、RACGAP1、BIRC5、KIF11 and CYP2B6 were prognosis key genes. And the prognosis analysis showed that the expressions of CDC6、PIK3R1、KIF11 and RACGAP1 were increased, and the expression of CENPE was decreased, which was closely related to prognosis of HCC. Conclusion: CDC6、CENPE、PIK3R1、KIF11 and RACGAP1 may be closely related to poor prognosis of HCC, and can be used as molecular biomarkers for future research of HCC prognosis.
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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
Abstract174)      PDF(pc) (1061KB)(401)       Save
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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Dihydromyricetin ameliorates chronic social defeat stress induced cognitive and affective disorder in mice
WANG Le, LI Bi-rong, XIAO Zhi-yong, ZHAO Jin-long, YU Xu-dong
CJAP    2019, 35 (6): 496-500.   DOI: 10.12047/j.cjap.5852.2019.108
Abstract77)      PDF(pc) (1230KB)(397)       Save
Objective: To investigated the effects of dihydromyricetin on cognitive and affective disorders induced by chronic social defeat stress and its possible mechanism in mice. Methods: C57BL/6J mice were randomly divided into control group (Control), chronic social defeat stress group (CSDS) and chronic social defeat stress + DHM group (CSDS+DHM) (14 mice in each group). The mice received chronic social defeat stress and were injected with DHM or vehicle intraperitoneally. A part of mice were subjected to (10 mice of each group) novel object recognition test (NOR), Y maze test, open field test (OFT), social interaction test (SIT), forced swimming test (FST) and tail suspension test (TST). The other mice (4 mice of each group) were decapitated and the expression levels of SIRT1 in hippocampus were detected by Western blot. Results: Compared with the control group, the learning and memory of the CSDS group were reduced significantly, the anxiety level was increased significantly, the immobility time in TST and FST was increased significantly, and the SIRT1 protein level in hippocampus was reduced significantly (P< 0.05 or P< 0.01); Compared with the CSDS group, the learning and memory of the CSDS + DHM group were improved significantly, the anxiety level of the mice was reduced significantly, and the immobility time in TST and FST was reduced significantly. The protein level of SIRT1 in hippocampus was increased significantly (P< 0.05 or P< 0.01). Conclusion: DHM ameliorates the cognitive impairment, anxiety like behavior and depression like behavior of mice induced by CSDS and up-regulates the expression of SIRT1 protein.
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Effects of puerarin on rats with type 2 diabetes mellitus
YANG Fei, DONG Xin-xin, GUO Yun
CJAP    2019, 35 (4): 355-358.   DOI: 10.12047/j.cjap.5786.2019.075
Abstract152)      PDF(pc) (921KB)(393)       Save
Objective: To investigate the therapeutic effects of puerarin on rats with type 2 diabetes mellitus (T2DM). Methods: T2DM models were established by high fat and high glucose feeding combined with a one-time intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). Then the rats were randomly divided into normal group, model group, metformin group (MET, 40 mg/kg), puerarin low-dose group, medium-dose group and high-dose group (40, 80, 160 mg/kg), n=10. After the model was successfully established, rats were treated with corresponding drug intervention by intragastrical administration for 4 weeks. The body weight and fasting blood glucose (FBG) were measured per week, and blood samples were collected 24 h after the last administration, and serum levels of blood glucose, serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholestrol (HDL-C), serum enzyme activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), serum creatinine (SCr), and blood uric acid (UA) were measured. Results: As compared with normal group, the body weight was decreased after 4 weeks-intervention in the model group, and the levels of FBG, TC, TG, LDL-C, ALT, AST, BUN, SCr and UA were all increased,while HDL-C level was decreased (P<0.05). As compared with model group,the body weight was increased after 4 weeks-intervention in metformin group and puerarin groups, and the levels of FBG, TC, TG, LDL-C, ALT, AST, BUN, SCr and UA were decreased (P<0.01); meanwhile, HDL-C level was increased significantly (P<0.05). Conclusion: Puerarin can reduce the weight loss of T2DM rats, decrease the blood lipid and blood glucose levels of T2DM rats, which can be used to control T2DM.
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Effects of endurance exercise on synaptic plasticity in cerebral cortex of aged rats and related regulatory mechanism
LIU Wen-feng, LIU Shao-peng, FU Rang, WANG Zhi-yuan, KUANG He-yu, XIA Yan, TANG Chang-fa
CJAP    2019, 35 (4): 339-345.   DOI: 10.12047/j.cjap.5803.2019.072
Abstract177)      PDF(pc) (1553KB)(388)       Save
Objective: To understand and analyze the rules of endurance exercise on the cerebral cortex adaptive mechanism in aged rats. Methods: In this study, 3-month-old (n=20), 13-month-old (n=24) and 23-month-old (n=24) specific-pathogen free (SPF) male Sprague-Dawley Rat (SD) rats were divided into young (Y-SED), middle-aged (M-SED) and old-aged (O-SED) sedentary control group, and the corresponding Y-EX, M-EX and O-EX in the endurance exercise runner group. The 10-weeks of regular moderate-intensity aerobic exercise intervention were carried out in the endurance exercise runner group. The exercise mode is treadmill exercise (slope 0), and the exercise intensity gradually increases from 60%~65% of the maximum oxygen consumption (V·O2max) to 70%~75%, and the exercise time is 10 weeks. Hematoxylin and eosin (HE) staining was used to detect age-related morphological changes. The expressions of superoxide dismutase(SOD) and brain-derived neurotrophic factor (BDNF) and the expressions of synapsin 1 (SYN1) and Ca2+/calmodulin- dependent protein kinases IIα (CaMK IIα) / AMP-activated protein kinase α1(AMPKα1) / mammalian target of rapamycin (mTOR) pathway -related genes were detected. Results: The cerebral cortex structure of the rats in each group showed age-related aging changes, the expression of SOD in the cortex showed a gradual decline, the expression of BDNF showed an age-increasing trend, and the expression levels of SYN1 and CaMK IIα were increased with age. The changes in AMPKα1 and SirT2 and IP3R, AKT1 and mTOR mRNA levels were increased slightly in middle-aged rats and decreased in aged rats. Compared with the rats in each sedentary control group, the nucleus of the cerebral cortex was tightly arranged and the number of nuclei observed under the microscope was increased significantly in each exercise group. Exercise promoted the expressions of SOD, BDNF and synaptophysin SYN1 in the cortex of rats, and the expression levels of SOD and BDNF in aged rats were up-regulated significantly (P< 0.01). The expression level of SYN1 in rats was up-regulated significantly (P<0.05) in the young and aged rats. The expression of CaMK IIα in the cortex of middle-aged and aged rats was up-regulated (P<0.01), while the expression level of CaMK IIα in young rats was down-regulated (P<0.01). Exercise could up-regulate the expression level of AMPKα1 in the cortex of young rats (P< 0.05), but not in middle-aged and old-age rats. Exercise could up-regulate the expression of SirT2 in the cortex of rats in all age groups (P<0.05). Exercise up-regulated the expression of phosphoinositide 3-kinase (IP3R)/ protein kinase B 1(AKT1) /mTOR in the cortex of rats, among which young IP3R was significantly up-regulated (P<0.01) in the young group, mTOR was significantly up-regulated in young and middle-aged group (P<0.01), and mTOR was also significantly up-regulated in the aged group (P<0.05). Conclusion: Endurance exercise up-regulates BDNF expression, regulates CaMKIIα signaling, activates AMPK signaling pathway and IP3R / AKT1 / mTOR signaling pathway, and improves synaptic plasticity in the cortex.
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Effects of Ringer’s solution with different concentrations of alcohol on biphasic compound action potentials of frog sciatic nerve trunk
HUANG Zhi-hua, WEI Pei-jian, JIANG Ling, CHEN Sui, CHENG Bi-hong, LIN Ying, WU Lin-geng, XU Qiu-xiong, WU Shao-wei, WANG Hai-yan, SHEN Jian-xin
CJAP    2019, 35 (3): 232-238.   DOI: 10.12047/j.cjap.5750.2019.050
Abstract237)      PDF(pc) (1864KB)(384)       Save
Objective: To quantitatively investigate the effects of Ringer’s solution with different concentrations of alcohol (1%~80%) on biphasic compound action potentials (AP) from frog sciatic nerve trunk, and their recoveries from alcohol effects. Methods: Individual segments of frog sciatic nerve trunk with a length of 6 to 8 cm were prepared. Ringer’s solution with different concentrations of alcohol (0%, 1%, 2%, 4%, 8%, 16%, 32%, 48%, 64% and 80%) was applied onto the segment of the trunk between the stimulus and ground electrodes via an agent reservoir which was newly armed in a nerve trunk shielded chamber for 5 minutes. The nerve trunk was respectively electro-stimulated to generate the biphasic compound AP which was recorded using the experimental system of BL-420F. This was followed by 5 times washout plus 5 min administration with Ringer’s solution before recovery recording of AP. Results: Compared to normal Ringer’s solution, Ringer’s solution with alcohol at ≤4% did not have dramatic impacts on the AP amplitude and conduction velocity, while Ringer’s solution with alcohol at ≥8% there was significant decrease in these two parameters. Ringer’s solution with alcohol at the conentrations of 16%, 32% and ≥48% could prevent a small proportion (30%), a large proportion (90%) and all (100%) of sciatic nerve trunks, respectively, from generating AP. Washout with normal Ringer’s solution after alcohol application at the concentration of ≤32%, AP could totally recover to normal status. While alcohol at the concentration of 48%, 64% and 80%, the probabilities to regenerate APs were 90%, 40% and 0%, and the AP amplitudes were decreased to 60%, 36% and 0%, respectively. After washout, AP conduction velocity showed no difference with alcohol at the concentration of ≤8% when compared with that before washout, while it could not be recovered to normal under alcohol at ≥16%. Conclusion: Ringer’s solution with different concentrations of alcohol exerts different effects on biphasic compound AP amplitude and conduction velocity. Hopefully, our findings could be helpful for the alcoholic usage and its recovery from alcoholic damage.
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CJAP    2019, 35 (3): 279-282.   DOI: 10.12047/j.cjap.5774.2019.058
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Effects of metformin on depressive behavior in chronic stress rats
LI Gai-fen, ZHAO Ming, ZHAO Tong, CHENG Xiang, FAN Ming, ZHU Ling-ling
CJAP    2019, 35 (3): 245-249.   DOI: 10.12047/j.cjap.5775.2019.052
Abstract256)      PDF(pc) (1199KB)(376)       Save
Objective: To detect the effects of metformin on the depressive-like behaviors in rats. Methods: Forty male SD rats were randomly divided into four groups: control group (CON group), metformin group (MET group), model group (CUMS group), model + metformin group (CUMS + MET group), 10 rats in each group. Chronic unpredictable mild stress (CUMS) method was used to establish rat depression model in three weeks. After the model was established successfully, two metformin groups were intraperitoneally injected with metformin (100 mg/kg), while the control group and the model group were injected with the same amount of saline once a day for two weeks. After that, the changes of weight gain, sucrose water preference experiment, forced swimming test, tail suspension immobility test and open field test were detected. The morphological changes of hippocampus were observed by Nissl staining. Results: Compared with the control group, the weight gain of rats in CUMS group was significantly slowed down (P<0.05), the sucrose preference rate and the spontaneous activity were significantly reduced (P<0.05), and the immobility time in forced swimming and tail suspension immobility test was significantly prolonged (P<0.05), and the morphological structure of hippocampus was changed, which confirmed the success of CUMS depression model. Compared with CUMS group, metformin treatment had no significant effect on body weight of rats, but it could significantly improve sucrose water intake, immobility time and spontaneous activity of CUMS depression model rats (P<0.05), and improve the abnormal morphological changes of hippocampus in CUMS rats. Conclusion: Metformin has a therapeutic benefit against CUMS-induced depression, which provides a new treatment for patients with diabetes mellitus complicated with depression.
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Effects of mice macrophages on skeletal muscle cells under high glucose treatment
LUO Wei, AI Lei, WANG Bo-fa, WANG Li-ying, GAN Yan-ming, ZHOU Yue
CJAP    2020, 36 (2): 124-129.   DOI: 10.12047/j.cjap.5901.2020.028
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Objective: To study the effects of mice macrophages on myogenic differentiation and insulin sensitivity of skeletal muscle cells under high glucose condition. Methods: C2C12 myoblasts and RAW264. 7 macrophages were co-cultured in transwell and treated with 60 mmol/L glucose. They were randomly divided into single culture control group (SC group, n=12), co-culture control group (CC group, n=12), single culture high glucose group (SH group, n=12) and co-culture high glucose group (CH group, n=12). Cell morphology was observed by phase contrast microscope. C2C12 were collected after 1 and 3 days of co-culture. Cell viability was measured by CCK-8. Embryonic myosin heavy chain (E-MHC) and glucose transporters 4 (GLUT4) protein expressions were detected by immunofluorescence. The expressions of myogenic factor 5 (Myf5), myogenic determination gene (MyoD) and myogenin gene were detected by real-time PCR. 2-(N-(7-nitrobenz-2-oxa-13-diazol-4-yl) amino)-2-deoxyglucose (2-NBDG) assay was used to detect the cellular basis and insulin-stimulated glucose uptake. Results: Under normal glucose concentration, this co-culture with RAW264. 7 promoted C2C12 myotube formation, E-MHC protein expression (P<0. 01), MyoD and myogenin gene expressions (P< 0. 05), insulin-stimulated 2-NBDG uptake (P<0. 05), and basic GLUT4 level (P<0. 05). High glucose stimulation inhibited myotube formation, myogenic regulatory factor gene expression, 2-NBDG uptake and GLUT4 expression in C2C12 (P<0. 05). When co-cultured with C2C12 under high glucose treatment, compared with co-culture control group and high glucose group, cell activity, E-MHC protein expression, myogenic regulator gene expressions, 2-NBDG uptake and GLUT4 protein expression were significantly decreased (P<0. 05). Conclusion: Co-culture with RAW264. 7 promotes myogenic differentiation and increases insulin sensitivity in C2C12, but this effect is reversed under 60 mmol/L glucose treatment, which inhibits myogenic differentiation and induces insulin resistance.
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Exogenous spermine alleviates myocardial injury induced by hyperglycemia by inhibiting golgi stress
FAN Yu-qi, WANG Yue-hong, SHAO Yi-ying, SHAO Xiao-ting, HU Jing, XU Chang-qing, WEI Can
CJAP    2019, 35 (5): 385-389.   DOI: 10.12047/j.cjap.5795.2019.082
Abstract136)      PDF(pc) (1314KB)(370)       Save
Objective: To investigate whether Golgi stress (GAS) is involved in diabetic cardiomyopathy (DCM) and whether myocardial protection of exogenous spermine is associated with regulation of GAS. Methods: Sixty Wistar rats were randomly divided into normal control group (Control), diabetic group (T1D, STZ 60 mg/kg intraperitoneal injection) and spermine group (T1D+Sp, spermine 5 mg/(kg·d) intraperitoneal injection) for 12 weeks. H9C2 rat cardiomyocytes were randomly divided into control group (Control, 10% FBS-DMEM culture), high glucose group (HG, 10% FBS-DMEM+40 mmol/L glucose) and spermine group (HG+Sp, 10% FBS-DMEM+40 mmol/L glucose+5 μmol/L spermine). Rat serum creatine kinase isoenzyme (CK-MB) and cardiac troponin T (cTnT) were detected by ELISA; Golgi protein GOLPH3, GM130 and Cleaved Caspase3 protein expressions were analyzed using Western blot; immunofluorescence was used to detect GOLPH3 cell localization. Results: In the animal model, compared with the normal group, myocardial ultrastructural damage was obvious,the blood glucose, the serum myocardial enzymes CK-MB and cTnT, the expressions of GOLPH3 and cleaved caspase3 were increased or up-regulated significantly,meanwhile, the body weight,the ejection fraction (EF) and the expression of GM130 were decreased or down-regulated markedly in the diabetic group. In the cell model, similar results were obtained. Immunofluorescence showed stress fragmentation of Golgi apparatus. Exogenous spermine treatment could significantly alleviate the above mentioned damages. Conclusion: Golgi stress occurs in diabetic cardiomyopathy (DCM), and myocardial protection of exogenous spermine is associated with a reduction in Golgi stress (GAS).
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Effects of curcumin on liver fibrosis induced by cholestasis in mice
WU Chen, QIU Yu-bao, SUN Xue-qian, CHEN Dan, WU Ya-xian, PANG Qing-feng
CJAP    2019, 35 (5): 468-472.   DOI: 10.12047/j.cjap.5796.2019.102
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Objective: To investigate the protective effects of curcumin on bile duct ligation(BDL)-induced liver cholestasis in mice, so as to provide a new treatment strategy for liver fibrosis. Methods: Forty-two healthy adult male BALB/c mice were randomly divided into sham group (n =6), sham+curcumin group (n=6), BDL treatment group (n=10), BDL+curcumin group(n=10), BDL+curcumin+ZnPP group (n=10). Seven days after BDL operation, the sham operation + curcumin group and the BDL+ curcumin group were treated with curcumin at the dose of 30 mg/kg by intraperitoneal injection once a day for 7 days.The mice in BDL+ curcumin +ZnPP group were treated with curcumin (30 mg/kg) and ZnPP (50 μmol/kg) by intraperitoneal injection once a day for 7 days. For the sham group and the BDL group, mice were treated with equal-volume saline daily by intraperitoneal injection. After 14 days of BDL, the plasma and liver tissues were collected, the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured. The pathological changes of liver tissue and liver fibrosis were observed, and the protein expression of HO-1 in liver tissue was detected. Results: Compared with the sham group, mice in the BDL group had enlarged liver gallbladder and the serum levels of ALT and AST were increased significantly (P<0.05). Meanwhile, the results of Sirius red staining and qRT-PCR of pro-fibrosis related genes showed collagen deposition in the liver, and immunohistochemistry of macrophages and neutrophils showed inflammatory cell infiltration in the liver. Compared with the BDL group, the serum levels of ALT and AST in the curcumin treatment group were decreased significantly (P<0.05), collagen deposition and inflammatory cell infiltration were improved, and HO-1 expression was increased (P<0.05) after curcumin treatement. In the curcumin treatment group, the protective effect of curcumin on liver injury could be reversed by HO-1 active inhibitor ZnPP. Conclusion: Curcumin can improve liver inflammation and fibrosis caused by BDL, and this protective effect is related to the regulation of HO-1 activity by curcumin.
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