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  • Table of Content
      28 September 2018, Volume 34 Issue 5 Previous Issue    Next Issue
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    Effects of TGF-β1 on the expression and secretion of cytokines induced by Aβ1-42 in hippocampal neuron and microglia co-cultures
    FANG Xiao-xia, ZHOU Yi, SUN Gao-lin, QIU Yi-hua, PENG Yu-ping
    CJAP. 2018, 34 (5): 385-388.   DOI: 10.12047/j.cjap.5668.2018.088
    Abstract   PDF (1151KB) ( 328 )
    Objective: To investigate the neuroprotective effects of transforming growth factor beta 1(TGF-β1) on the expression and secretion of cytokines induced by Aβ1-42 in hippocampal neurons and microglial co-cultures. Methods: Hippocampal neurons and microglia obtained from SD rat were co-cultured. TGF-β1 was applied on day 5 after the neurons and microglia co-cultures were incubated at the concentrations of 5 or 20 ng/ml, Aβ1-42 was added 1 h following TGF-β1 application at a concentration of 5 μmol/L. They were incubated for 72 h and then assessed for further studies. Western blot analyses were employed to examine the expression of inducible nitric oxide synthase (iNOS); Real-time PCR and ELISA were used to detect the mRNA expression and secretion of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and insulin-like growth factor-1 (IGF-1). Results: In the hippocampal neuron-microglia co-cultures, Aβ1-42 induced upregulation of iNOS, TNF-α and IL-1β, downregulation of IGF-1. TGF-β1 pretreatment ameliorated the pro-inflammatory effects caused by Aβ1-42. Conclusion: TGF-β1 significantly inhibits the increase in inflammatory cytokines and the decrease in neurotrophic factor which are caused by Aβ1-42-induced microglia activation.
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    The improvement effect of scFv17 on the gait of triple-transgenic mice
    WANG Ting, CAO Yue, HU Meng-ming, ZHANG Xiu-min, DONG Xue-fan, QI Jin-shun, ZHANG Jun, YANG Wei
    CJAP. 2018, 34 (5): 389-395.   DOI: 10.12047/j.cjap.5656.2018.089
    Abstract   PDF (2118KB) ( 177 )
    Objective: To observe the gait changes of Alzheimer's disease PS1M146V/APPswe/tauP301L triple-transgenic (3xTg-AD) mice and to investigate the improvement effect of single chain variable domain antibody fragment 17 (scFv17) on the gait.Methods: In the present study, a selection of 6-month-old 3xTg-AD mice (n=18) and C57BL/6 wild-type mice (n=24) was performed. First, we observed their gait changes and found that the gait of 12-month-old 3xTg-AD mice was severely damaged. Then, the two groups of mice were randomly divided into four groups:WT+PBS(n=12), WT+scFv17(n=12), 3xTg-AD+PBS(n=9) and 3xTg-AD+scFv17(n=9). The gait behavior test and pathological test were performed after 12 weeks'continuous administration of scFv17 (1.5 mg/kg) or an equal volume of PBS (0.01 mol/L) by nasal gavage twice a week. Results: Compared with the same month old wild type mice, the rear track width of 12 month old 3xTg-AD mice was increased(P<0.01), swing time percent was decreased (P<0.01), stance time percent was increased(P<0.01), so the ability of movement coordination and balance was seriously damaged. ScFv17 could improve the coordination and balance ability of 12 month old 3xTg-AD mice(P<0.01). The morphological structure of 3xTg-AD mice cerebellar Purkinje cells was improved. The treatment of scFv17 increased the Nissl body number of the cerebellar Purkinje cells of 3xTg-AD mice (P<0.01). scFv17 reduced the amyloid β protein (Aβ) plaques in the cerebellar cortex of 3xTg-AD mice (P<0.01), and scFv17 reduced the intracellular neurofibrillary tangles (NFT) of the cerebellar Purkinje cells of the 3xTg-AD mice (P<0.01). Conclusion: The coordination and balance ability of 3xTg-AD mice was significantly impaired. ScFv17 can improve gait behaviour in the 3xTg-AD significantly.The mechanism may be related to the improvement of the structure and protein function of cerebellar Purkinje cells, and the eliminating of the Aβ plaques and the neurofibrillary tangles.
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    Placenta-derived mesenchymal stem cells with HLA-G positive expression induce Treg in vitro
    CUI Gui-yu, BAI Jian, MIAO Lan-ying, LIN Da-yong, LIU Hong, LI Ya-li, LIU Xi-cheng
    CJAP. 2018, 34 (5): 396-400.   DOI: 10.12047/j.cjap.5621.2018.090
    Abstract   PDF (1144KB) ( 258 )
    Objective: To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro. Methods: placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4+ T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was accounted. Results: After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (P<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4+ T lymphocytes were cultured for 48 h, the ratio of CD4+CD25+Foxp3+Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (P<0.01). Conclusion: Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4+CD25+Foxp3+Treg in vitro.
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    Effects of endothelial progenitor cells on proliferation and biological function of hepatic stellate cells under shear stress
    CHEN Xin-xin, ZHANG Xiao-yun, DING Yu-zhen, LI Xin, GUAN Xiu-mei, LI Hong, CHENG Min, CUI Xiao-dong
    CJAP. 2018, 34 (5): 404-407.   DOI: 10.12047/j.cjap.5624.2018.092
    Abstract   PDF (1169KB) ( 334 )
    Objective: To investigate the effects of endothelial progenitor cells (EPCs) under shear stress on the biological function such as proliferation, adhesion, migration, apoptosis and expression of α-smooth muscle actin (α-SMA), collagen-I and collagen-Ⅲ of hepatic stellate cells (HSCs). Methods: HSCs and EPCs were inoculated into the upper and lower layers of the co-culture chamber respectively and co-incubated for 24 hours. Then, 12 dyne/cm2 shear stress was applied to EPCs cells for another 24 hours. After that, proliferation, adhesion, migration and apoptosis of HSCs were detected by cell counting kit-8 (CCK-8) kit, cell adherent assay, Boyden cell migration assay and flow cytometry respectively. Fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of alpha -SMA, collagen I and collagen-Ⅲ in HSCs. Results: Under shear stress, EPCs ecological niche could obviously inhibit the proliferation, adhesion and migration of HSCs, promote the apoptosis of HSCs, and down-regulate the mRNA and protein expression of collagen-I, collagen-Ⅲ in HSC cells. Conclusion: Under shear stress, EPCs ecological niche could inhibit the fibrosis development of HSCs to a certain extent.
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    The effect of Yiqi Wenyang Huoxue Huatan Fang on hypoxia-hypercarbia induced pulmonary hypertension and its mechanism
    ZHANG Cong-cong, ZHANG Jing-jing, CHEN Jun-hao, WU Yuan-ling, HUANG Dan-na, DAI Yong-yue, WANG Wan-tie
    CJAP. 2018, 34 (5): 408-413.   DOI: 10.12047/j.cjap.5626.2018.093
    Abstract   PDF (3495KB) ( 262 )
    Objective: To investigate the effect of Yiqi Wenyang Huoxue Huatan Fang (YWHHF) on alleviating hypoxia-hypercarbia pulmonary hypertension by inhibiting endothelial-mesenchymal transition (EndoMT) via BMP-7/Smads pathway. Methods: Fifty male healthy SD rats of clean grede, weighting (180~220) g, were randomly divided into 5 groups (n=10):normoxia group (N), hypoxia-hypercarbia group (HH); YWHHF high dose group (YH), middle dose group (YM) and low dose group (YL). The rats in N group were kept in normal oxygen environment, the remaining four groups were intermittently exposed to hypoxia-hypercarbia environment (9%~11% O2, 5%~6% CO2) for 4 weeks, 6 days a week, 8 hours per day. The rats in YH, YM, YL groups were received YWHHF gavage in a dosageof 0.6, 0.3, 0.15g/kg respectively (3 ml/kg),the rats in N and HH groups were received equal volume of normal saline. After 4 weeks, the mean pulmonary arterial pressure(mPAP) was detected,the right ventricular free wall and left ventricle plus ventricular septum were isolated to determine the right ventricular hypertrophy index. Lung ultrastructural changes were surveyed under an electronic microscopy, the changes of pulmonary artery structure surveyed by immunofluorescence, the mRNA levels of alpha-smooth muscle actin (α-SMA)、platelet endothelial cell adhesion molecule-1 (CD31)、bone morphogenetic protein-7 (BMP-7)、drosophila mothers against decapentaplegic protein1/5/8 (Smad1/5/8) were detected by RT-PCR, and the protein levels of α-SMA、CD31、BMP-7、p-Smad1/5/8 and Smad1/5/8 were detected by Western blot. Results: Compared with N group, mPAP and the right ventricular hypertrophy index were increased,some significant injuries also were discovered under microscopic observation,the mRNA and protein expression of α-SMA was increased, and the mRNA expressions of CD31、BMP-7、Smad1/5/8 were decreased in the other four groups, the protein expressions of CD31、BMP-7、p-Smad1/5/8 were decreased(P<0.05). Compared with HH group, the above changes in YH、YM、YL groups were all improved (P<0.05). Conclusion: YWHHF can inhibit EndoMT to alleviate pulmonary hypertension, and the mechanism may be related to the promotion of the expression of BMP-7/Smads pathway.
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    Effects of astragaloside-IV on the expression of inflammatory factor and proliferation of glomerular mesangial cells induced by angiotensin Ⅱ
    XIONG Li, LYU Meng-juan, DOU De-yu, MA Yu-hong
    CJAP. 2018, 34 (5): 414-417.   DOI: 10.12047/j.cjap.5660.2018.094
    Abstract   PDF (1224KB) ( 248 )
    Objective: To study the effects of astragaloside-IV (As-IV) on the expression of inflammatory factor and proliferation of glomerular mesangial cells (GMCs) induced by angiotensin Ⅱ(AngⅡ). Methods: The in vitro model of diabetic nephropathy(DN) was mimic by angiotensin Ⅱ (10-6mol/L)inducing GMCs injury. Then the GMCs were treated with As-IV at different concentrations(25,50,100 μmol/L)for 48 hours. The proliferation of GMCs was detected by MTT. The level of reactive oxidative species (ROS) was measured by flow cytometry. The expression of monocyte chemoattractant protein-1(MCP-1) protein in supernatant was detected by enzyme- linked immunosorbent assay (ELISA). The expression of transforming growth factor-β1(TGF-β1) in GMCs was measured by Western blot. Results: Compared with model group, the proliferation of GMCs was inhibited in As-IV group. As-IV decreased the level of intercellular ROS, down-regulated the secretion of MCP-1 and the expression of TGF-β1 proteins. Conclusion: As-IV could inhibit cell proliferation and inflammatory factors expression on GMCs induced by AngⅡ.
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    Effects of laser stimulation with different power on oxygen consumption rate of C2C12 myoblasts in mice
    FENG Yu, CHENG Ze-peng, SUN Peng, SHI Reng-fei
    CJAP. 2018, 34 (5): 418-421.   DOI: 10.12047/j.cjap.5667.2018.095
    Abstract   PDF (1077KB) ( 198 )
    Objective: To investigate the effects of 650 nm laser irradiation on cell oxygen consumption rate in C2C12 myoblasts following different doses. Methods: C2C12 cells were irradiated with 650 nm laser(λ=650 nm, p=5 mW) with energy densities of 0, 0.4, and 0.8 J/cm2. Cell oxidative function was measured by oxygen consumption rate kit. Protein expression of myogenic determination factor (MyoD), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), mammalian target of rapamycin (mTOR) and its phosphorylation were detected by Western blot. Results: Compared to the control group, the expression levels of MyoD, PGC-1α protein were increased and cell oxygen consumption rate was promoted in the low dose group(P<0.05). MyoD and PGC-1α protein expressions were also increased(P<0.05), the ratio of mTOR and its phosphorylationwere decreased significantly in the high dose group(P<0.05). Conclusion: 650 nm laser irradiation that dose is 0.4 J/cm2 enhances cell oxidative function, it related to that proper dose laser irradiation promoted the expression of PGC-1a protein.
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    Simvastatin prevented myocardium of diabetes rats from apoptosis through inhibition of oxidative stress
    LI Fan-lu, WAN Xin, WANG Xi, LIU Xin, WU Ya-li, CHEN Huan-zhen, CUI Xiang-li
    CJAP. 2018, 34 (5): 422-426.   DOI: 10.12047/j.cjap.5653.2018.096
    Abstract   PDF (5913KB) ( 257 )
    Objective: To investigate the protective effects and the possible mechanisms of simvastatin on myocardial injury induced by diabetes.Methods: Twenty-four SD rats (180~220)g were randomly divided into control group (control, n=8) and modeled groups(n=16), the modeled groups were injected with streptozotocin intraperitoneally to induce diabetes. Then the modeled rats were randomly divided into diabetes mellitus group (DM group, n=8) and diabetes mellitus + simvastatin group (DM+S group, n=8). Rats in DM+S group were treated with simvastatin at the dose of 40 mg/(kg·d)by gavage for 4 weeks, and the other two groups were treated with the same amount of saline. At the end of experiments, the heart tissues were collected for further observation. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in heart tissues were measured by spectrophotometry; HE staining of rat heart slides was used to observe the pathological changes; TUNEL assay was used to determine the apoptosis index of myocardial cells in each groups; The distribution of p53 in the heart tissues was evaluated by immunohistochemistry; Western blot was used to detect the expressions of p53, p53-phospho-serine 15, Bax and Bcl-2 in the heart tissues. Results: ①Compared with control group, the content of malondialdehyde (MDA) was increased while the activity of superoxide dismutase (SOD) was decreased significantly in DM group (P<0.01). After simvastatin administration, the activity of SOD was increased and the content of MDA was decreased significantly (P<0.01). ② HE staining results showed that the myocardial cells in the DM group were disorganized, with unclear morphological structure and a large number of inflammatory cells infiltration. Compared with DM group, the myocardial morphology in DM+S group was improved significantly. ③TUNEL staining results showed that the apoptosis index of myocardial cells in DM group was increased significantly compared with that of control group, and the apoptosis index was decreased significantly after the treatment of simvastatin (P<0.01).④ Immunohistochemistry showed that compared with control group,the expression of p53 in DM group was increased significantly, and was expressed in both cytoplasm and nucleus, while the expression of p53 in DM+S group was decreased and the expression of p53 in nucleus was decreased significantly (P<0.01). ⑤ The results of Western blot showed that the expression levels of p53, p53-phospho-serine15 and Bax were higher than those in control group, and the expression of Bcl-2 was lower than that in control group (P<0.01). After simvastatin administration, the expression levels of p53,p53-phospho-serine 15 (P<0.01) and Bax were decreased significantly (P<0.05) and the expression of Bcl-2 was increased (P<0.05). Conclusion: Simvastatin exerted protective effects on myocardial injury caused by diabetes through improving the abnormal morphological changes of diabetic myocardium, alleviating oxidative stress and inhibiting apoptosis of myocardial cells. The mechanism is related to the regulation of apoptosis pathway mediated by p53.
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    Effects of irbesartan on Notch1 signaling pathway in diabetic rats with myocardial injury
    ZHANG Heng, TAO Min, KANG Pin-fang, GUO Jian-lu, XUAN Ling, GAO Qin, TANG Bi, LI Lu-jia, WANG Hong-ju
    CJAP. 2018, 34 (5): 427-431.   DOI: 10.12047/j.cjap.5674.2018.097
    Abstract   PDF (1410KB) ( 246 )
    Objective: To investigate the effects and mechanisms of irbesartan on myocardial injury in diabetic rats, and to analyze the changes of Notch1 signaling pathway in it. Methods: Thirty rats were randomly divided into four groups:normal control group (CON, n=6), high calorie group (HC, n=6) and diabetes mellitus group (DM, n=9), irbesartan + diabetes group (Ir + DM, n=9). After modeling 8 weeks later, the body weight ratio and left ventricular weight index were measured and the serum levels of triglyceride (TG) and total cholesterol (TC) were measured by automatic biochemical analyzer. The changes of superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium of rats were determined by the kit and the expressions of B-cell lymphoma-2 (Bcl-2) and Bcl-2 assaciated X protein (Bax) protein in myocardium were detected by immunohistochemistry. The expressions of Notch1, Hes-1 and jagged-1 in myocardium of rats were detected by Western blot. Results: Compared with CON group, the levels of heart weight/body weight (H/B), left ventricular weight index(LVWI) and fasting blood glucose(FBG) in HC group were not significantly changed, while the levels of blood lipids, MDA and Bax were increased significantly, and the expressions of SOD, Bcl-2 and Notch1, Hes-1 and Jagged-1 were decreased. Compared with HC group, the levels of H/B, LVWI, FBG, MDA and Bax in DM group were increased significantly, and the levels of SOD, Bcl-2 and Notch1, Hes-1 and Jagged-1 were decreased. The expression of H/B, LVWI, Notch1, Hes-1 and Jagged-1 in Ir+DM group were increased, but there was no significant difference between the other indexes. The H/B and LVWI in Ir + DM group were significantly lower than those in DM group, the levels of blood lipid and blood glucose did not change significantly, but the incidence of oxidative stress and apoptosis was reduced. While Notch1, Hes-1, Jagged -1 protein expressions were increased. Conclusion: Diabetes can induce myocardial injury, and irbesartan has myocardial protective effects through activation of Notch1.
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    Protective effect of Ginkgo biloba extract on paracetamol-induced acute hepatic injury in mice
    ZHANG Quan-shu, WANG Xiang-peng, XIE Yan-ni, WU Lu-lu, LIU Hong
    CJAP. 2018, 34 (5): 432-435.   DOI: 10.12047/j.cjap.5655.2018.098
    Abstract   PDF (2462KB) ( 229 )
    Objective: To investigate the protective effects of Ginkgo biloba extract(GBE) on paracetamol(APAP)-induced acute hepatic injury in mice and its mechanism. Methods: Thirty mice were randomly divided into control group, model group, GBE low, medium and high-dose(50,100,and 200 mg·kg-1)groups,with 6 mice in each group. All mice except control group were administered with APAP(300 mg/kg)for one time by intraperitoneal injection. The mice in GBE low, medium and high-dose groups were intragastric administered with GBE for 2 d consecutively, then samples were harvested for analysis. The appearance and pathology of liver were observed. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum and the levels of superoxide dismutase (SOD), myeloperoxidase(MPO), glutathione (GSH) and malondialdehyde (MDA) in hepatic tissue were measured. Western blot was used to detect the protein expressions of Nrf2 and HO-1. Results: Compared with control group, in model group, the appearance and pathology of liver were bad, the levels of ALT,AST,TNF-α and IL-6 in serum were increased significantly(P<0.01),the levels of GSH and SOD were decreased while the levels of MDA and MPO were increased in hepatic tissue(P<0.01), the expressions of Nrf2 and HO-1 were increased in hepatic tissue(P<0.05). Compared with model group, in GBE groups, the appearance and pathology of liver were improved, the levels of ALT,AST,TNF-α and IL-6 in serum were decreased significantly(P<0.01), the levels of GSH and SOD were increased while the levels of MDA and MPO were decreased in hepatic tissue(P<0.01), the expression of Nrf2 and HO-1 were increased in hepatic tissue(P<0.05). The high-dose of GBE possessed the most obvious treatment effect among them. Conclusion: GBE may play a protective role in APAP-induced acute hepatic injury through Nrf2/HO-1 pathway.
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    Analysis of high-dose dexamethasone in different cycles on efficacy and safety of newly diagnosed ITP
    XIAO Yue, LI Tang-fei
    CJAP. 2018, 34 (5): 436-440.   DOI: 10.12047/j.cjap.5673.2018.099
    Abstract   PDF (919KB) ( 279 )
    Objective: To compare the efficacy and safety of high-dose dexamethasone in treating new-diagnosed primary immune thrombocytopenia (ITP) in monocycle, di-cycle, tri-cycle. Methods: Divided by the ratio of 1:1:1, 93 newly diagnosed patients were randomly accepted monocycle (Group A:dexamethasone 40 mg once a day, from day1 to day4), dicycle (Group B:dexamethasone 40 mg once a day, from day 1 to day 4, day 15 to day 18), tri-cycle (Group C:dexamethasone 40 mg once a day, from day 1 to day 4, day 15 to day 18, day 29 to day 32) of high-dose dexamethasone treatment. Its efficacy and safety on the patients in three groups were compared. Results: Ninety-three newly patients with new-diagnosed ITP were divided into Group A, B, and C, 31 patients in each group. In terms of short-term benefits, there was no statistically significant difference among the 7th and 14th day complete response rate after end of treatment. However, there was statistically significant difference after the end of treatment on the 7th day response rate (41.9% vs 70.0% vs 90.0%, P<0.01) and the 14th day response rate (16.1% vs 36.70% vs 63.3%, P< 0.01); in terms of long-term benefits, there was no statistically significant difference among the 120-day response rate, the complete response rate within the treatment on the 60th, 90th and 120th day and the relapse rate at 90th and 120th day; however, there was statistically significant difference among the 60- day response rate (10.0% vs 26.6% vs 53.3%, P<0.01), 90-day(0.0% vs 13.3% vs 30.0%, P<0.01) and 60-day relapse rate(88.9% vs 73.3% vs 46.7%, P<0.01). Mostly of the treatment-related adverse reactions in the three groups were mild, and most patients are tolerable. Conclusion: Although the complete response rate of ITP patients did not improved by increasing the cycle of high-dose dexamethasone, but improved response rate in three months, and adverse reactions were tolerable, which could be used as a reference for clinical use.
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    Effects of chloride channels on hemolysis induced by puerarin injection
    JIANG Feng-qi, YUAN Ying, XIANG Zuo-jun, HUANG Pan-rui, WU He, WANG Shi-qi, WANG Juan, PENG Hui, LUO Hai
    CJAP. 2018, 34 (5): 441-444.   DOI: 10.12047/j.cjap.5681.2018.100
    Abstract   PDF (1857KB) ( 248 )
    Objective: To observe the effects of blocking and activating chloride channels on hemolysis induced by puerarin injection in rabbits and to investigate the roles of chloride channels in hemolytic reaction induced by puerarin injection. Methods: Rabbit erythrocyte suspension was incubated with different concentrations of puerarin injection(0.75, 1.5, 3, 6, 12 mg/ml) at 37oC for 6 hours. The cell imaging system was employed to observe whether puerarin injection induced hemolysis. The hemolysis rate was detected by microplate reader and flow cytometry. Effects of activating and closing chloride channels on the hemolysis induced by puerarin injection were explored. Results: Puerarin injection could induce the hemolysis of rabbit erythrocytes in vitro. In the range of 1.5 mg/ml~12 mg/ml, puerarin injection could induce hemolysis in a concentration-dependent manner (n=3, P<0.01). The chloride channel blockers tamoxifen (20 μmol/L) and ATP (10 mmol/L) significantly inhibited the hemolysis induced by puerarin injection (n=3~5, P<0.01). Application of low concentration ATP (50 μmol/L) to activate the chloride channel significantly increased puerarin injection induced hemolysis (n=4, P<0.01). Conclusion: The hemolytic effect of puerarin injection is dose-dependent in vitro, and the activation of chloride channel is closely related to the hemolysis induced by puerarin injection.
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    Effects of aerobic exercise combined with chlorella pyrenoidos of disintegrated cell wall on some indicators of lipid metabolism in rats with high-fat diet
    ZHANG Jing, MIAO Gang, CAO Jian-min, ZHOU Hai-tao, NIU Yan-long, ZHANG Yue, REN Yi, BAO Xin-yue, XING Ya-wen
    CJAP. 2018, 34 (5): 445-449.   DOI: 10.12047/j.cjap.5688.2018.101
    Abstract   PDF (927KB) ( 278 )
    Objective: To study the effects of aerobic exercise combined with chlorella pyrenoidos of disintegrated cell wall on the lipid metabolism in rats with high-fat diet. Methods: Fifty-five male Wistar rats were subjected to adaptive feeding for 4 days and weight-free swimming training for 3 days, 20 min/d. After eliminating 5 rats that were not suitable for swimming training, the other rats were randomly divided into 5 groups according to their body weight:control group (C group), high fat diet group (H group), high-fat diet + chlorella group(HC group), high fat diet + aerobic exercise group (HM group), high fat diet + chlorella + aerobic exercise group (HMC group), 10 in each group. The HM and HMC group were subjected to 60 min/d swimming training for 6 weeks with non-weight-bearing. Group C were fed regular diet. The other groups were fed with high-fat diet, the rats in group HC and HMC were intragastrically treated with chlorella pyrenoidos of disintegrated cell wall at the dose of 3.9 g/(kg·d), the volume was 5 ml/kg, and the other groups are given equivalent saline. The Lee's index and biochemical indexes of blood and liver were measured after 6 weeks. Results: Compared with group C, Lee's index, serum levels of free fatty acids(FFA), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-c), liver FFA and interleukin-10 (IL-10) were increased significantly (P<0.01), the serum level of high-density lipoprotein cholesterol (HDL-c) was decreased significantly (P<0.01) in group H. Compared with group H, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-c, liver FFA and IL-10 were decreased significantly (P<0.05 or P<0.01), serum level of HDL-c was increased significantly (P<0.05 or P<0.01) in group HC, HM and HMC. Compared with group HC and HM, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-c, liver FFA and IL-10 were decreased significantly (P<0.05), serum level of HDL-c was increased significantly (P<0.05) in group HMC. Conclusion: Aerobic exercise and chlorella pyrenoidos of disintegrated cell wall can improve lipid metabolism in rats with high-fat diet and reduce the lipid toxicity caused by obesity. Joint intervention is more effective than single intervention.
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    Study on 4 weeks medium intensity aeraobic exercise induce the difference expression of the proteomics in rat atrial muscle
    PENG Yong, SHI Shao-rong, HUANG Si-min, GUO Yuan-pan, XU Zhe, WAN Li-li, LEI Xiong
    CJAP. 2018, 34 (5): 450-456.   DOI: 10.12047/j.cjap.5683.2018.102
    Abstract   PDF (1500KB) ( 234 )
    Objective: To investigate the effect of moderate-intensity aerobic exercise on the differential expression of rat atrial muscle Proteomics and genes, which provide research basis for the rehabilitation of chronic cardiovascular diseases and exercise -induced cardiac remodeling research. Methods: Twenty male SD rats were randomly divided into control group and experimental group (n=10) according to body weight. Rats in the experimental group were trained (6 days per week),which lasted for 4 weeks of moderate-intensity aerobic exercise at a rate of 24 m·min-1 for 40 min (load intensity equivalent to 60%~70% VO2max). The proteins were separated by two dimensional gel electrophoresis, and the tandem time-of- flight mass spectrometer technique was used to identify 13 candidate target protein spots. The expression levels of these 13 protein spots were up-regulated more than 5 times or down -regulated to below 1/5. The mRNA of six target proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results: By software analysis, the experimental group compared with the control group, there were 8 protein points which their expression reduced more than 4/5 and 5 protein points up-regulated more than 5 times, 13 proteins were identified by mass spectrometry protein spots, the final identification results acquired 8 proteins and a unknown protein of molecular mass 54 KDa, such as:pyruvate dehydrogenase E1α1, mitochondrial aconitate hydratase, protein disulfide isomerase A3, methylmalonic acid semialdehyde dehydrogenase, mitochondrial dihydrolipoic acid dehydrogenase, isovaleryl coenzyme A dehydrogenase, glutathione synthetase, mitogen-activated protein kinase 3 and so on. Compared with the control group, the mRNA expression of methylmalonic acid semialdehyde dehydrogenase in the atrial muscle of rats was decreased after 4 weeks of moderate aerobic exercise (P<0.05), the mRNA expression levels of mitochondria Ⅱ lipoic acid dehydrogenase, protein disulfide isomerase A3, mitochondrial aconitate hydratase and glutathione synthetase were decreased (P>0.05); The mRNA expression level of isopentenyl-CoA dehydrogenase was increased (P>0.05). The results indicated that the mRNA expression level was not completely consistent with the changes in mass spectrometry identification results. Conclusion: The 4 weeks moderate-intensity aerobic exercise induced ignificant changes of rats atrial muscle protemics. The majority of the 13 identified target proteins in this experiment are energy metabolism enzymes. The majority of the expression of the target protein and the mRNA expression in the atrial muscle is inconsistent and different. Exercise may affect the regulation of gene transcription or downstream translation and modification of these target proteins, resulting in the change of differential expression.
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    Early cardiac injury in patients with obstructive sleep apnea
    LI Jie-ru, GAO Xiu-hua, HAN Ju-qiang, WANG Gui-ying, KANG Li-ying, JI En-sheng
    CJAP. 2018, 34 (5): 457-461.   DOI: 10.12047/j.cjap.5707.2018.103
    Abstract   PDF (929KB) ( 203 )
    Objective: To evaluate the early cardiac injury caused by obstructive sleep apnea (OSA) before the development of cardiovascular symptoms of OSA. Methods: Ninety-two patients without any known cardiovascular disorders who underwent polysomnography (PSG) were enrolled in the study. Subjects were divided into mild, moderate, and severe OSA groups by their apnea hypopnea index (AHI), and 25 healthy individuals were identified as controls. After PSG examination, fasting blood samples for the evaluation of N-terminal pro-brain natriuretic peptide (NT-proBNP) and heart-type fatty acid binding protein (h-FABP) were collected in the morning, and left ventricular(LV) functions were assessed by using echocardiographic methods. Thirty moderate and severe OSA patients were treated with continuous positive airway pressure respectively (CPAP).Results: The levels of h-FABP and NT-proBNP were obviously higher in all OSA groups than those in the control group (P<0.01), and were positively correlated with AHI (P<0.01). The Em/Am values of all OSA groups and E/A values of the moderate and severe OSA groups were significantly reduced (P<0.01). The difference in Em/Am values among the groups was statistically significant (P<0.01). Compared with those before treatment, h-FABP and NT-BNP levels in serum of OSA patients after CPAP treatment were significantly reduced (P<0.01), and Em/Am and E/A values were significantly increased (P<0.01). Conclusion: Left ventricular diastolic dysfunction and early myocardial microtrauma are major manifestations of early heart damage in patients with OSA. CPAP therapy could significantly improve early cardiac damage in OSA patients.
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    Effect of Beclin-1 shRNA on hypoxia-induced autophagy in SH-SY5Y cells
    LU Na, LI Cheng-zhang, LUO Xiao-qiu, YANG Kun-li
    CJAP. 2018, 34 (5): 464-469.   DOI: 10.12047/j.cjap.5661.2018.105
    Abstract   PDF (1403KB) ( 346 )
    Objective: To observe the effect of hypoxia on autophagy in Beclin-1-knockdown SH-SY5Y cells by constructing a stable transfected SH-SY5Y cell lines of silencing Beclin-1 gene. Methods: Beclin-1shRNA lentiviral vector and negative control lentiviral vector were constructed; the vector was transfected into SH-SY5Y cells; then the expression of Beclin-1 mRNA was detected by RT-PCR, the level of Beclin-1 protein was detected by Western blot. CCK-8 method was used to determine the effect of Beclin-1 knockdown on the viability of SH-SY5Y cells. Next, the blank control, negative control and transfected cells were cultured under 21% normoxia and 5% hypoxia conditions. The expression of LC3 protein in each group was detected by Western blot and the autophagic bodies were observed by electron microscopy. Results: Beclin-1 shRNA significantly inhibited the expression of Beclin-1 mRNA and protein in SH-SY5Y cells; after silencing Beclin 1 gene, the survival rate of Beclin-1 shRNA group cells was no different from that of negative control (NC) group. After 5% hypoxia treatment, compared with NC group, the ratio of LC3Ⅱ/LC3Ⅰand the number of autophagy bodies were all decreased in Beclin-1 shRNA group. Conclusion: Beclin-1 knockdown SH-SY5Y cell lines and negative control cell lines were successfully established. Lentivirus-mediated Beclin-1 shRNA has no effect on the viability of SH-SY5Y cells, but can inhibit hypoxia-induced autophagy.
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    The receptor-dependent and non-receptor-dependent mechanism of melatonin activated BKCa channels in middle cerebral artery myocyte
    CHEN Yu, XU Zhao-xia, ZHANG Hui-rong, WU Ying, SHI Li-jun
    CJAP. 2018, 34 (5): 470-475.   DOI: 10.12047/j.cjap.5675.2018.107
    Abstract   PDF (2157KB) ( 262 )
    Objective: To investigate the mechanisms through which myocyte large-conductance Ca2+-activated K+ (BKCa) channels mediate the vasodilation effects of melatonin on cerebral arteries (CAs). Methods: Middle cerebral arteries (MCA) were obtained from 8-week-old male Wistar rats after anaesthetized. Middle cerebral arterial smooth muscle cells were enzymatically isolated. Whole cell recording mode of patch clamp technique was used to measure the current density of BKCa channel and voltage-gated potassium (KV) channel before and after adding melatonin. Currents density of melatonin on BKCa channels with melatonin receptor inhibitor 2-phenyl-N-acetyl (luzindole) was recorded using whole cell recording mode and open probability (Po) was recorded using single-channel attached recording mode. The conductance (G) and average open time (To) and off time (Tc) of the BKCa channel were detected before and after the addition of melatonin in the internal-outward mode. Results: ① Melatonin markedly increased the whole-cell BKCa channel current density but not the voltage-gated potassium (KV) channel current density. ② Luzindole (1 μmol/L) greatly suppressed melatonin-induced increase of BKCa channel current density. ③ The Po of BKCa channel was significantly increased by melatonin (100 μmol/L) under cell attached recording mode, which was markedly inhibited by luzindole (1 μmol/L). ④ In inside-outside recording mode, melatonin (1 μmol/L, 100 μmol/L) reduced both To and Tc of BKCa channel, and Tc was reduced much more than To. Conclusion: Melatonin mediates vasodilation of MCA through the activation of BKCa channels via both melatonin receptor dependent and independent mode.
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    The effects of anthocyanin on chronic inflammatory pain induced by complete Freund's adjuvant and its mechanism
    WANG Jing, CHEN Ze-feng, HE Chun-xia, WANG Li-li, ZHENG Jia-hui, ZHANG Hao, WANG Zhan-lei, LU Ya-ping
    CJAP. 2018, 34 (5): 476-480.   DOI: 10.12047/j.cjap.5671.2018.106
    Abstract   PDF (1143KB) ( 301 )
    Objective: To investigate the effects and mechanisms of anthocyanin from Ligustrum vicaryi on chronic inflammatory pain induced by complete Freund's adjuvant. Methods: Thirty male SD rats were randomly divided into three groups (n=10):normal saline control group (NS), chronic inflammatory pain model group(Mod, injected with complete Freund's adjuvant(CFA) 100 μl to the left hind leg), anthocyanin treatment group(Ant, dosed with anthocyanins (90 m), mechanical pain threshold (MPT), and left toe volume in each group were measured before modeling and 1,3,5,7,9,11,13 days after operation. Antioxidant indexes in serum were mensurated by spectrophotometer, and the total capsaicin receptor (TRPV1) and phosphorylated capsaicin receptor (p-TRPV1) in hippocampus were detected by Western blot. Results: In comparison with controls, HPT and MPT were improved (P<0.05),toe swelling was reduced(P<0.05), the serum level of SOD was increased (P<0.01), while the levels of MDA and NO were decreased (P<0.05), the ratio of P-TRPV1/TRPV1 protein was depressed in Mod rat hippocampal region treated with anthocyanin. Conclusion: The results show that anthocyanins has an analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism may be related to the improvement of antioxidant capacity and the reduction of TRPV1 phosphorylation.
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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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