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  • Table of Content
      28 May 2019, Volume 35 Issue 3 Previous Issue    Next Issue
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    ORIGINAL ARTICLES
    Injury of rat blood vessels caused by acute ozone exposure and its mechanism
    YANG Hu, LI Ning, HAN Jie, ZHU Chen-li, TIAN Lei, LIN Ben-cheng, XI Zhu-ge, LIU Xiao-hua, CHU Nan
    CJAP. 2019, 35 (3): 193-198.   DOI: 10.12047/j.cjap.5748.2019.042
    Abstract   PDF (997KB) ( 236 )
    Objective: To investigate the vascular damage effects and possible mechanism of acute exposure to ozone (O3) in male Wistar rats. Methods: One hundred and twenty male Wistar rats were randomly divided into six groups, 20 in each group. The experimental animals were placed in a gas poisoning cabinet, the control group was exposed to filtered air, and the treatment group was exposed to ozone at concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, 2.0 ppm, and 4.0 ppm, respectively, for 4 hours. Arterial blood pressure data were obtained by PC-lab medical physiological signal acquisition system. Blood rheology indicators and blood biochemical indicators were detected by Tianjin Dean Diagnostic Laboratory. Serum endothelin-1 (ET-1), homocysteine (HCY), von Willebrand factor (vWF), 8-hydroxydeoxyguanosine (8-OhdG), interleukin (IL-6) and tumor necrosis factor alpha (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) microplate assay. Oxidative stress indicators superoxide dismutase (SOD) activity and malondialdehyde (MDA) were determined by xanthine oxidase method, thiobarbituric acid (TBA) method, reduced glutathione (GSH) and nitric oxide (NO) were tested by using microplate colorimetry. Paraffin sections were prepared from thoracic aorta tissue, and vascular structure was observed by HE staining. Results: Acute exposure to 0.12 ppm ozone could cause a significant increase in arterial systolic blood pressure (SBP). Exposure to different concentrations of ozone could cause a significant increase in plasma viscosity, and the K value of the ESR equation was significantly increased in the 1.0 ppm ozone exposure group. Both the relative and reduced viscosities were significantly reduced at ozone concentrations of 0.5 ppm and 4.0 ppm, while the red blood cell deformation index was increased significantly at ozone concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, and 2.0 ppm. Acute ozone exposure resulted in the decrease of total cholesterol content. The content of high-density lipoprotein cholesterol (HDL-C) was significantly reduced in the 0.12 ppm ozone exposure group. When the ozone concentration was higher than 1.0 ppm, the body may also had an inflammatory reaction (increased TNF-α) and oxidative stress (increased MDA, decreased GSH). Acute exposure to ozone could lead to elevated levels of ET-1 in the blood, with significant differences in the 4.0 ppm concentration group, while HCY levels were decreased firstly and then increased, reaching the highest in the 1.0 ppm concentration group. No obvious pathological changes were observed in the thoracic aorta. Conclusion: Acute ozone exposure can affect arterial blood pressure, blood rheology and cholesterol metabolism in rats. The possible mechanism is that ozone exposure leads to inflammatory reaction and oxidative stress reaction, causing vascular endothelial function damage, and vascular endothelial cells increase with ozone exposure concentration.
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    Expression and significance of hypoxia-inducible factor-1α and vascular endothelial growth factor and receptor 2 in stage 3 pressure injury of rats
    WANG Xiao-hui, CHEN Xiao-ping, WANG Hong-ping, PAN Ying-ying, JIANG Li-ping
    CJAP. 2019, 35 (3): 199-203.   DOI: 10.12047/j.cjap.5730.2019.043
    Abstract   PDF (1135KB) ( 184 )
    Objective: To analyze the expression and relationship of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and kinase insert domain receptor (KDR) in local skin tissues of pressure injury and investigate the possible mechanism of stage 3 pressure injury refractory wound.Methods: Forty male SD rats were randomly divided into normal control group, compressed 3 d, 5 d, 7 d, and 9 d groups. Stage 3 pressure injury animal model were established by magnet compression. The morphology of skin was observed by HE staining. The expression of VEGF was detected by immunohistochemistry. The expression levels of HIF-1α, VEGF and KDR protein in skin tissue were detected by Western blot. One-way analysis of variance and LSD test were performed on the data. Results: ①The HE results showed that compared with the normal control group, the epidermis of the compressed group was gradually thickened, the number of blood vessels was decreased, the collagen arrangement disordered and inflammatory cells infiltration were increased. ②Immunohistochemical results showed that the expression of VEGF protein in the 3 d group was significantly higher than that in the normal control group (P<0.01). The expression of VEGF protein in the skin tissue of 5 d, 7 d and 9 d groups was lower than that in normal control group (P<0.05). WB results were consistent with immunohistochemistry results. ③WB results showed that the expression of HIF-1α in the skin tissues of the rats in 3 d, 5 d and 7 d groups was higher than that in the normal control group (P<0.01 or P<0.05). The expression of KDR protein was lower than that of the normal control group (P<0.05 or P<0.01). Conclusion: HIF-1α mediated reduction of VEGF and KDR protein expression and decreased tissue angiogenesis may be one of the important causes of chronic dysfunction of stage 3 pressure injury.
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    The effects of Xiaotan Huayu Liqiao formula on mesenteric artery function in rats exposed to chronic intermittent hypoxia
    YU Fu-yang, YANG Sheng-chang, GUO Ya-jing, ZHAO Ya-shuo, LIU Si-wen, JI En-sheng
    CJAP. 2019, 35 (3): 204-208.   DOI: 10.12047/j.cjap.5725.2019.044
    Abstract   PDF (1215KB) ( 200 )
    Objective: To investigate the effects of Xiaotan Huayu Liqiao formula (the Chinese Medicine) on mesenteric artery function in rats exposed to chronic intermittent hypoxia (CIH), and to explore the related mechanism. Methods: Forty-eight male SD rats were randomly divided into four groups as Normoxia, CIH, Formula+CIH and formula group. Rats were exposed to normoxia in the Normoxia and Formula group, or intermittent hypoxia in CIH or Formula+CIH group. Xiaotan Huayu Liqiao formula was given at 24g/kg by intragastric administration before intermittent hypoxia exposure. The pathological changes of mesenteric artery were determined by HE staining, and the relaxation of mesenteric artery (induced by acetylcholine(ACh) and L-arginine(L-Arg)) was recorded by microvascular ring technique. Serums of all rats were collected (0 d and 21 d) and the content of NO was detected by ELISA. The levels of endothelial nitric oxide synthase (eNOS) and p-eNOS were measured by Western blot. Results: Compared with Normoxia group, the mesenteric arterial endothelial injury and media thickening were observed and the relaxation of mesenteric artery was significantly reduced in rats exposed to CIH. The level of NO in serum and the ratio of p-eNOS/eNOS were also decreased in the CIH group. Xiaotan Huayu Liqiao formula administration improved the pathologic changes and dilatation function of mesenteric artery, increased the levels of NO and p-eNOS. Compared with Normoxia group,all the results were not observed significant difference in Formula group. Conclusion: Xiaotan Huayu Liqiao formula increased the bioavailability of NO, and ameliorated the CIH induced mesenteric artery function injury.
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    Effects of apple polyphenols on monocrotaline-induced pulmonary vascular remodeling in rats and its mechanism
    ZHANG Shu-hao, SHAO Si-ming, CHEN Fang-zheng, ZHU Jing, CHEN Luo-wei, WANG heng, XIANG Xin-hui, YUAN Lin-bo
    CJAP. 2019, 35 (3): 209-214.   DOI: 10.12047/j.cjap.5769.2019.045
    Abstract   PDF (1812KB) ( 294 )
    Objective: To investigate the effects of apple polyphenols on pulmonary vascular remodeling in rats with pulmonary arterial hypertension and its mechanism. Methods: Rats were randomly divided into 4 groups:control (Con) group, monocrotaline (MCT) group, apple polyphenol (APP) group,monocrotaline + apple polyphenol (MCT+APP) group. In Con group, rats received a subcutaneous injection of physical saline. In APP group, rats received intraperitoneal injection of 20 mg/kg APP, every other day. In MCT group, rats received a single subcutaneous injection of MCT(60 mg/kg). In MCT+APP group, rats received subcutaneous injection of 60 mg/kg MCT followed by an intraperitoneal injection of 20 mg/kg APP every other day. All the disposal lasted 3 weeks. Then the PAH-relevant indicators, such as mean pulmonary artery pressure(mPAP), pulmonary vascular resistance(PVR), right ventricular hypertrophy index (RVHI) ,wall thickness (WT%) and wall area (WA%) were tested. After that, the inflammatory pathway related indicators, such as interleukin1(IL-1),interleukin1(IL-6), tumor necrosis factor α(TNF-α), cyclooxygenase 2(COX-2) and myeloperoxidase(MPO) in pulmonary tissue and free intracellular Ca2+ in pulmonary smooth muscle cell(PASMC), content of eNOS and NO in endothelial cells were determined. Results: Compared with the control group, the levels of mPAP, PVR, RVHI, WA%, WT%, and IL-1, IL-6, TNF-α, COX-2, MPO in tissue and the expression of Ca2 + in PASMC of MCT group were increased significantly, while the contents of eNOS and NO in endothelial cells were decreased significantly (P<0.05). Compared with the MCT group, the apple polyphenol treatment could improve the above mentioned situation, and the COX-2 and Ca2+ indicators of the apple polyphenol treatment group were decreased significantly (P<0.05). Conclusion: MCT can increase COX-2 expression and intracellular Ca2+ in pulmonary artery smooth muscle cells, decrease the contents of eNOS and NO in endothelial cells, while apple polyphenols can significantly inhibit these effects.
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    The exploration of aerobic power and energy expenditure of Chinese rugby players
    FU Tao, YANG Yue-wen, WU Peng, LIU Guan-jun
    CJAP. 2019, 35 (3): 215-218.   DOI: 10.12047/j.cjap.5760.2019.046
    Abstract   PDF (1048KB) ( 252 )
    Objective: To explore aerobic power and energy expenditure of high level rugby players in China, which provide experimental basis for accurate training and nutritional strategy in match-play. Methods: Eighteen master rugby players were selected as research subjects. The parameters such as VO2max, lactic aicd threshold (LT) and modify conconi test were measured respectively. The differences of energy were compared between the forward and the defender. The data were analyzed by independent sample t test. Results: The VO2max(42.05±3.69 ml/min·kg-1) of rugby players was poorer. The VO2max of the forward was 38.83±3.52 (ml/min·kg-1), and that of the defender was 47.31±3.17 (ml/min·kg-1),and there was significant difference between the forwards and the defenders (P<0.05). The LT of the defenders was obviously higher than that of the forwards. Modifier conconi test had a high correlation (r = 0.772) with VO2max. The average energy consumption in the first half of the game was about(276.94±18.08)kcals, the second half was(225.58±22.86)kcals, and the second half was less than the first half (P<0.05).Conclusion: The aerobic power is different between the forwards and the defenders. The power of aerobic of Chinese players is weaker than that of the foreign rugby players.
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    Therapeutic effect of massage on denervated skeletal muscle atrophy in rats and its mechanism
    WAN Xiao-feng, TANG Cheng-lin, ZHAO Dan-dan, AN Hui-yu, MA Xiang, QIAO Tong-xi
    CJAP. 2019, 35 (3): 223-227.   DOI: 10.12047/j.cjap.5726.2019.048
    Abstract   PDF (1689KB) ( 320 )
    Objective: To investigate the therapeutic effects of massage on denervated skeletal muscle atrophy in rats and its mechanism. Methods: Forty-eight male SD rats were randomly divided into model group (n=24) and massage group (n=24). Gastrocnemius muscle atrophy model was established by transecting the right tibial nerve of rat. On the second day after operation, the gastrocnemius muscle of the rats in the massage group was given manual intervention and the model group was not intervened. Six rats were sacrificed at the four time points of 0 d, 7 d, 14 d and 21 d. The gastrocnemius of the rats were obtained and measured the wet mass ratio after weighing. Cross-sectional area and diameter of the muscle fiber were measured after HE staining. The relative expressions of miR-23a, Akt, MuRF1 and MAFbx mRNA were tested with qPCR. Results: Compared with 0 d, the wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle showed a progressive decline in the model group and massage group. The wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle in the massage group were higher than those in the model group on 7 d, 14 d and 21 d (P<0.05, P<0.01). Compared with 0 d, the expressions of MuRF1, MAFbx and Akt mRNA were increased first and then were decreased in the model group and massage group. The expression of MuRF1 mRNA in massage group was lower than that in model group on 7 d and 21 d (P<0.05, P<0.01). The expression of MAFbx mRNA in massage group was lower than that in model group on 7 d, 14 d and 21 d (P<0.01, P<0.05, P<0.01). The expression of Akt mRNA in massage group was higher than that in model group on 7 d, 14 d and 21 d (P<0.05, P<0.01). Compared with 0 d, the expression of miR-23a mRNA was increased in the model group and massage group on 21 d, and the expression of miR-23a mRNA in massage group was higher than that in model group (P< 0.05). Conclusion: Massage can delay the atrophy of denervated skeletal muscle. The mechanism may be related to up-regulation of the expression of miR-23a and Akt mRNA, down-regulation of the expressions of MuRF1 and MAFbx mRNA, inhibition of protein degradation rate, and reduction of skeletal muscle protein degradation.
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    Effects of ginsenoside-Rg2 on mechanical allodynia, heat hyperalgeia, depressive state of rats with chronic sciatic nerve constriction injury
    ZHANG Qiao-lian, LI Shuang-ying, LI PING
    CJAP. 2019, 35 (3): 228-231.   DOI: 10.12047/j.cjap.5763.2019.049
    Abstract   PDF (925KB) ( 213 )
    Objective: To investigate the effects of ginsenoside-Rg2 on mechanical allodynia, heat hyperalgeia, depressive state of rats with chronic sciatic nerve constriction injury. Methods: Fifty SD rats were randomly divided into 5 groups: blank control group (Normal, normal + saline),sham operation group (Sham, sham operation + saline),chronic constriction injury of the sciatic nerve group (CCI, CCI + saline),ginsenoside-Rg2 low dose group (CCI + Rg2 5 mg/kg), and ginsenoside-Rg2 high dose group (CCI + Rg2 10 mg/kg).After the CCI model was established,drug were injected into the abdominal cavity through the syringe once a day,for 14 consecutive days.The mechanical shrinkage foot reflex threshold (MWT) and thermal withdrawal latency(TWL) were determined at 1 d before the operation and at 1,3,5,7,10 and 14 d after the operation.Light-dark transition test, forced swimming test were determined at 1 d before the operation and at 14 d after the operation.Results: Compared with the sham group, the MWL and TWL of the CCI rats were decreased significantly (P<0.01), time in the light compartment and number of transition were decreased (P<0.01), the immobility time in FST was also prolonged significantly (P<0.01). At 14 days after CCI operation, the MWL and TWL of the ginsenoside-Rg2 groups were increased significantly (P<0.01), time in the light compartment and number of transition were also shortened significantly (P<0.01), the immobility time in FST was also shortened significantly (P<0.01). Conclusion: Intraperitoneal injection of ginsenoside-Rg2 can inhibit the mechanical and thermal pain sensitivity of CCI rats,and can relieve depressive state.
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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
    Abstract   PDF (1864KB) ( 384 )
    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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    The electrophysiological response of chorda tympani nerve to taste stimuli in rats with conditioned taste aversion to saltiness
    LI La-mei, LIAO Ying-ying, JIANG En-she
    CJAP. 2019, 35 (3): 239-244.   DOI: 10.12047/j.cjap.5768.2019.051
    Abstract   PDF (1284KB) ( 192 )
    Objective: To explore the characteristic changes of the peripheral chorda tympanic nerve (CT) electrophysiological responses to salty stimulus and other taste stimuli in rats with the conditioned taste aversion to saltiness. Methods: Fourteen adult SD male rats were divided into a conditioned taste aversion to salty group (CTA) and a control group (Ctrl) (n=7/group). On the first day of the experiment, rats were given a 0.1 mol/L NaCl intake for 30 min, then, the rats in CTA and Ctrl groups were injected intraperitoneally with 2 ml of 0.15 mol/L LiCl and the same amount of saline respectively. On day 2, 3 and 4, the 30 min consumption of NaCl and distilled water was measured for both groups of rats. On the 4th day after the behavioral test of that day, CT electrophysiological recording experiments were performed on CTA rats and control rats. Results: Compared with the rats in Ctrl group, the electrophysiological characteristics of CT in CTA group rats did not change significantly the responses to the series of NaCl and other four basic taste stimuli (P>0.05). The amiloride, the epithelial sodium channel blocker, strongly inhibited the response of CT to NaCl in CTA and Ctrl group rats (P<0.01). Conclusion: The electrophysiological responses of CT to various gustatory stimuli do not significantly change in rats after the establishment of conditional taste aversion to the saltiness.
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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
    Abstract   PDF (1199KB) ( 376 )
    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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    The effect of procyanidin on periprosthetic osteolysis caused by TCP wear particles in the mouse calvaria and its mechanism
    LIN Kun, CHEN Jia-hao, FANG Ze-hao, YE Cheng-long, HAN Chao-jie, YAN Ming, FANG Jian, ZHANG Yun
    CJAP. 2019, 35 (3): 250-255.   DOI: 10.12047/j.cjap.5765.2019.053
    Abstract   PDF (1488KB) ( 227 )
    Objective: To investigate the protective effects of procyanidin on periprosthetic osteolysis caused by tricalcium phosphate (TCP) wear particles in the mouse calvaria and its mechanism. Methods: Forty-eight male ICR mice were randomly divided into sham group, TCP group, and procyanidin (0.2 mg/kg, 1 mg/kg, 5 mg/kg)-treated group (n=12). A periprosthetic osteolysis model in the mouse calvaria was established by implanting 30 mg of TCP wear particles onto the surface of bilateral parietal bones following removal of the periosteum. On the 2nd day post-operation, procyanidin (1 mg/kg, 5 mg/kg) was locally injected to the calvaria under the periosteum every other day. After 2 weeks, all the mice were sacrificed to collect the blood samples and the calvaria. Periprosthetic osteolysis and osteoclastogenesis in the mouse calvaria were observed by tartrate resistant acid phosphatase (TRAP) staining and HE staining. mRNA levels of TRAP, capthesin K, c-Fos and NFATc1 in the periprosthestic bone tissue were examined by real-time fluorescence quantitative PCR. Serum contents of total anti-oxidation capacity (T-AOC) and MDA, and superoxide dismutase (SOD) activity were determined by chemical colorimetry. Protein expressions of autophagic biomarkers such as Beclin-1 and LC-3 in periprosthetic bone tissue of the calvaria were examined by Western blot. Results: Compared with sham group, periprosthetic osteolysis, osteoclastogenesis, mRNA levels of TRAP, capthesin K, c-Fos and NFATc1, and serum MDA content were increased significantly in the TCP group (P<0.05), whereas serum T-AOC level and SOD activity were decreased. The protein expressions of Beclin-1 and LC-3, and the conversion of LC3-II from LC3-I were both up-regulated markedly in the mouse calvaria of TCP group (P<0.05). Compared with TCP group, osteolysis, osteoclastogenesis, mRNA levels of TRAP, capthesin K, c-Fos and NFATc1 and serum MDA content were decreased obviously in the procyanidine group (P<0.05), serum T-AOC level and SOD activity were increased, the expressions of Beclin-1 and LC-3, and the conversion of LC3-II from LC3-I were down-regulated obviously in the mouse calvaria of procyanidin group (P<0.05). Conclusion: Procyanidin has a protective effect of periprosthetic osteolysis caused by TCP wear particles in the mouse calvaia, its mechanism may be mediated by inhibition of oxidative stress and autophagy.
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    The effects of optical genetic techniques on new neurons through the Wnt/β-Catenin pathway
    XIA Tian-guang, ZHU Xu, WANG Jing-jing, WEI Meng-guang, LYU Fang-fang, CHEN Chong, LIANG Jun, JIANG Wei, SUN Qian, SUN Hong-Tao
    CJAP. 2019, 35 (3): 256-261.   DOI: 10.12047/j.cjap.5731.2019.054
    Abstract   PDF (1928KB) ( 227 )
    Objective: To investigate the effects of optical genetic techniques on new neurons through the Wnt/β-Catenin pathway. Methods: Neural stem cells (ESCs)were extracted from the cerebral cortex of fetal rat and transfected by lentivirus carrying DCX-ChR2-EGFP gene and the expression of DCX of newborn neurons differentiated from neural stem cells were observed. All cells were divided into 3 groups(n=9): control group, NSCs+EGFP and NSCs+ChR2 groups. The control group was normal cultured NSCs (NSCs group); the neural stem cells in NSCs+EGFP group were transfected with lentivirus carrying EGFP gene. The neural stem cells in NSCs+ChR2 group were infected with lentivirus carrying DCX-ChR2-EGFP gene. After 48 hours of lentivirus infection, 470 nm blue laser irradiation was performed for 3 consecutive days. NeuN+ positive cell density(the maturation of neural stem cells)and the ratio of NeuN+/Hoechst in each group were observed. Western blot was used to detect the expression levels of MAP2, NeuN, Neurog2, NeuroD1 and GluR2. Western blot was used to detect the expressions of β-catenin and TCF4 associated with Wnt/β-catenin signaling channel. Verapamil (100 μmol/L, L-type calcium channel blockers) and Dkk1 (50 μg/ml, β-catenin inhibitor) were used to treat stem cells of the NSCs+ChR2 group and then the expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were detected by Western blot. Results: After 3 days of 470 nm blue laser irradiation, NeuN+ positive cell density(the maturation of neural stem cells)and the ratio of NeuN+/Hoechst, the expression levels of the protein MAP2, NeuN, Neurog2, NeuroD1, GluR and the protein β-catenin and TCF4 associated with Wnt/β-catenin signaling channel detected by Western blot were significantly increased in the group of NSCs+ChR2, compared with NSCs and NSCs+EGFP groups. The expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were remarkably decreased after treated by verapamil and Dkk1 in the group of NSCs+ChR2. It was proved that the opening of ChR2 channel producing cationic influx promoted the maturation of neural stem cells and induced by the Wnt/β-catenin signaling pathway. Conclusion: Optical genetic promoted the maturation of newborn neurons through the Wnt/β-catenin signaling pathway.
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    Effects of Exendin-4 on the differentiation of neural stem cells from subventricular zone of adult mice in vitro
    ZHAO Fei, XU Hui-you, MA Ke, JIANG Ji-peng, ZHANG Jian, DAI Chen, JIN Ying, LI Ping, SUN Hong-tao, WANG Zhen-guo, CHEN Xu-yi
    CJAP. 2019, 35 (3): 262-267.   DOI: 10.12047/j.cjap.5848.2019.055
    Abstract   PDF (3642KB) ( 230 )
    Objective: To study the effect of exendin-4(Ex-4) on the differentiation of neural stem cells(NSCs) in adult mouse subventricular zone(SVZ)and its mechanism . Methods: NSCs in the SVZ were derived from 5-week C57BL/6J mice and the expression of nestin was detected by immunofluorescence. The cell morphology was observed after the cells treatmed with 100 nmol/L Ex-4 for 14 days.The expressions of nestin and glucagon-like peptide-1 receptor (GLP-1R) were detected by immunofluorescence. GLP-1R was knocked down by using shRNA and the study was divided into four groups: control group, Ex-4 group, GLP-1R knockdown group, GLP-1R knockdown + Ex-4 group. After treatment with 100 nmol/L Ex-4 for 14 d, β-tublin III and glial fibrillary acidic protein (GFAP) were labeled by immunofluorescence and then the proportion of β-tublin III positive cells were counted. Western blot was used to detect the activation of cAMP-response element binding protein (CREB) in NSCs. In order to further study the effects of Ex-4 on mitogen-activated protein kinase(MAPK) and phosphatidylinositol 3-hydroxy kinase (PI3K) pathways, the cells were pretreated with MAPK inhibitor U0126 at a concentration of 0.07 μmol/L for 30 min or PI3K inhibitor LY294002 at 50 μmol for 2 h, respectively. The study was divided into six groups: control group, Ex-4 group, U0126 group, U0126 + Ex-4 group, LY294002 group, LY294002 + Ex-4 group. The activation of CREB in each group was detected by Western blot. The experiment was repeated three times independently. Results: NSCs were successfully extracted from SVZ of C57BL/6J mice. Immunofluorescence showed that nestin and GLP-1R were positive in NSCs. Compared with the control group, the proportion of neurons differentiated from Ex-4 group was higher. The percentage of neurons in GLP-1R knockdown + Ex-4 group was basically the same as that in control group (P<0.01). The positive cells of beta-tublin III showed positive activation of GLP-1R and CREB. Western blot showed that CREB was significantly activated in the Ex-4 group, and knockdown of GLP-1R abolished its activation (P<0.01). U0126 did not affect Ex-4-mediated CERB activation, and LY294002 significantly reduced Ex-4-mediated CREB activation (P<0.01). Conclusion: Ex-4 promotes the differentiation of NSCs into neurons in SVZ of adult mice through GLP-1R receptor, which may be achieved through PI3K/CREB pathway.
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    The expression of calcium-sensing receptor in rats with acute myocardial infarction and its effect on cells apoptosis
    YUAN Hui, YANG Guo-hong, LI Shu, LI Li, SONG Gao-chen, XU Chang-qing, SUN Jian
    CJAP. 2019, 35 (3): 268-272.   DOI: 10.12047/j.cjap.5753.2019.056
    Abstract   PDF (2112KB) ( 272 )
    Objective: To investigate the change of calcium sensing receptor (CaSR) expression at different time in rat tissue with acute myocardial infarction (AMI) and its effect on cardiomyocyte apoptosis. Methods: The healthy Wistar rats were randomly divided into Sham and AMI groups, the rat myocardial infarction model was established by ligating left anterior descending coronary artery. The changes of cardiac morphology and hemodynamics were detected at 1, 2 and 4 weeks,respectively. The expressions of CaSR mRNA and protein in myocardial tissue were detected by RT-PCR and Western blot, respectively. The expressions of Bax, Bcl-2, caspase-3 and caspase-9 proteins were detected by Western blot. The serum levels of lactate dehydrogenase (LDH), creatine kinase (CK) activity and cardiac troponin (cTnT) were determined. The apoptosis of cardiomyocytes were tested by TUNEL staining. Results: Compared with the sham group, the expressions of CaSR mRNA and protein, the apoptosis index were increased significantly with the development of AMI (P<0.05). The ultrastructural damage of cardiomyocytes was serious; the levels of LVSP, +dp/dtmax and -dp/dtmax were decreased,while the levels of LVEDP was increased (P<0.05); In AMI group, the cTnT level, CK and LDH activities were all increased (P<0.05). With the development of myocardial infarction, the cTnT level and CK activity were gradually decreased, while the activity of LDH was not significantly changed. The expressions of promote apoptosis-related Bax, caspase-3 and caspase-9 were significantly increased, and the expression of inhibited apoptosis-related protein(factor)Bcl-2 was significantly decreased (P<0.05). Conclusion: With the development of myocardial infarction,the expressions of CaSR mRNA and protein,the apoptosis index in rat myocardial tissue were increased with time prolongation after AMI. The increased expression of CaSR is involved in rat myocardial infarction, which is related with apoptosis.
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    Effects of MD2 gene silencing on high glucose-induced proliferation inhibition, apoptosis and inflammation in rat cardiomyocytes
    LIN Zhong-min, CHEN Guo-rong, ZHANG Quan-bo, WANG Fang, XIANG Lan-ting, CAO Qiong-jie
    CJAP. 2019, 35 (3): 273-278.   DOI: 10.12047/j.cjap.5834.2019.057
    Abstract   PDF (1905KB) ( 248 )
    Objective: To investigate the effects of myeloid differentiation-2 (MD2) gene silencing on high glucose-induced proliferation inhibition, apoptosis and inflammation in rat cardiomyocytes. Methods: The immortalized rat cardiomyocyte cell line H9C2 were transfected with MD2 small interfering RNA (si-MD2) and negative control for 24 h, then stimulated with high glucose (HG) for 48 h. RT-qPCR was performed to detect the mRNA levels of MD2 and inflammatory factors TNF-α, IL-1β and IL-6. MTS and flow cytometry were used to evaluate cell proliferation, cell cycle and apoptosis rate. Western blot was used to detect protein expression levels and phosphorylation levels. Results: The mRNA and protein levels of MD2 in H9C2 cells were dramatically decreased after transfected with si-MD2 (P<0.01). After stimulation of high glucose, the mRNA levels of inflammatory factors, the cells in G0/G1 phase , the cell apoptosis rate and the protein level of cleaved Caspase-3 were significantly increased, while the cell proliferation ability was decreased (P<0.01). MD2 gene silencing antagonized the effects of high glucose on cell proliferation, cell cycle, cell apoptosis and the mRNA levels of TNF-α, IL-1β , IL-6(P<0.05). Western blot analysis showed that the phosphorylation levels of extracellular signal-regulated kinase(ERK1/2), P38 mitogen-activated protein kinase(P38 MAPK) and C-Jun N-terminal kinase(JNK) protein were increased significantly in H9C2 cells treated with high glucose, which could be reversed by silencing of MD2 (P<0.01). Conclusion: This study demonstrates that MD2 gene silencing reverses high glucose-induced myocardial inflammation, apoptosis and proliferation inhibition via the mechanisms involving suppression of ERK, P38 MAPK, JNK signaling pathway.
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    The effects of tectochrysin on prostate cancer cells apoptosis and its mechanism
    WANG Yu, KE Rui-jun, JIANG Pan-ruo, YING Jia-hao, LOU En-zhe, CHEN Jia-yu
    CJAP. 2019, 35 (3): 283-288.   DOI: 10.12047/j.cjap.5754.2019.059
    Abstract   PDF (1248KB) ( 225 )
    Objective: To investigate the effects of tectochrysin on prostate cancer cell line 22Rv.1 and reveal its molecular mechanism. Methods: Tectochrysin at the concentrations of 0~20 μg/ml was applied to 22Rv.1 cells and normal prostate cell RWPE-1. The proliferation activity of the cells was detected by MTS assay. Flow cytometry and hoechst 33342 staining were used to analyze the effects of drugs on cell apoptosis, death, cell cycle and nuclear type changes. LDH release test was used to analyze the cytotoxicity of the drug to 22Rv.1 cells. QPCR and Western blot were used to analyze the effects of the drug on the expressions of genes in 22Rv.1 cells. Finally, the tumor inhibited effect of the drug on the bearing tumor BALB/c mice were confirmed though anti-tumor experiment. Results: Tectochrysin could significantly inhibit the proliferation activity of 22Rv.1 cells and induced their apoptosis, and promoted the expressions of genes dr4, dr5, trail, p53, caspase-3, caspase-8, caspase-9, bid, bax and foxo3, inhibited the expressions of anti-apoptotic genes akt, pi3k and bcl-2. Conclusion: Tectochrysin can induce prostate cancer cells apoptosis through affecting TRAIL and PI3K/AKT signaling pathways, and has anti-prostate cancer effect.
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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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