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Liraglutide is an analog of glucagon⁃like peptide⁃1 and plays an important role in the treatment of diabetes. But the specific mechanism of liraglutide in improving the function of pancreatic β cells is still unclear. Therefore, high glucose (33 mmol/ L) was used to induce islet MIN6 cells for 48 hours to establish a high glucose injury model in this study. The CCK⁃8 assay was used to detect cell viability, and the results showed that the viability of MIN6 cells in the high glucose group decreased (P<0. 05) compared with the control group, and the cell viability increased after liraglutide treatment (P<0. 05). Using the mouse insulin detection kit and ATP content detection kit, we found that both the insulin re⁃ lease (P<0. 01) and ATP content decreased (P<0. 001) in the high glucose group compared with the control group, and after liraglutide treatment both insulin release (P < 0. 05) and ATP content (P < 0. 001) increased. We used the mitochondrial membrane channel pore (MPTP) fluorescence detection kit in MIN6 cells and found that the green fluorescence intensity of the high glucose group was significant⁃ ly decreased (P<0. 001) compared with the control group, and after liraglutide treatment the green fluo⁃ rescence intensity was significantly increased (P<0. 001). The DCFH⁃DA probe combined with flow cy⁃ tometry was used to detect the level of reactive oxygen species (ROS). We found that compared with the control group, the ROS level in the high glucose group was significantly increased (P<0. 001), and de⁃ creased by liraglutide treatment (P<0. 01). Intracellular malondialdehyde (MDA) contents, superoxide dismutase (SOD) and catalase (CAT), and lactate dehydrogenase (LDH) activities in the cell superna⁃ tant were measured, and the levels of MDA and LDH were significantly increased (P<0. 05), and the levels of SOD and CAT were significantly decreased (P<0. 01) in the high glucose group compared with the control group. After liraglutide treatment, the levels of MDA and LDH were decreased (P<0. 05), and the levels of SOD and CAT were increased (P<0. 05). The results of Western blotting showed that the expression of UCP2 was upregulated in the high glucose group (P<0. 01) compared with the control group, and after liraglutide treatment the expression of UCP2 decreased (P<0. 05). The results indica⁃ ted that liraglutide has significant effects in high⁃glucose induced mitochondrial damage, oxidative stress and insulin secretion in MIN6 cells, which will provide a theoretical basis for clinical utilization of liraglu⁃ tide.
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Genipin (Gen) is an important antioxidant that plays a crucial role in the process of intracellular resistance to oxidative stress. In order to study the effect of genipin on MIN6 cells injured by high glucose, the CCK-8 method was used to detect the cell survival rate. The cell viability of the high-glucose injury group decreased (P <0. 05). But after genipin acted on the cells injured by high glucose, the cell viability increased (P <0. 05). The mouse insulin detection kit and ATP content detection kit found that the insulin release in the high glucose injury group decreased (P < 0. 001) and the ATP content decreased (P <0. 001). After genipin was given, the insulin release increased (P <0. 01), ATP content increased (P <0. 01). The fluorescent probe DCFH-DA detected the intracellular reactive oxygen species (ROS) level and found that the ROS content in the high glucose-injury group was significantly increased (P <0. 01). After genipin was administered, ROS content decreased (P < 0. 05). Glutathione(GSH) / oxidized glutathione (GSSG), intracellular malondialdehyde (MDA), superoxide dismutase(SOD) and catalase (CAT) and lactate dehydrogenase (LDH) activity in the cell supernatant revealed that the GSH / GSSH ratio, SOD and CAT levels in the high glucose injury group decreased (P <0. 05), and the intracellular MDA and LDH levels were significant increased (P<0. 001) .After administration of genipin, the GSH / GSSH ratio, SOD and CAT levels increased (P <0. 01), MDA and LDH levels were significantly reduced (P <0. 01). Mitochondrial membrane potential (MMP) levels decreased in the highglucose injury group (P <0. 01). After genipin acted on the cells injured by high glucose, the MMP level increased (P < 0. 05). Western blot detected uncoupling protein 2 (UCP2), antioxidative proteinsglutathione reductase (GR) and glutaredoxin 1 (Grx1) contents. The results showed that UCP2 contents in the high glucose injury group increased (P <0. 01) and the content of oxidized protein decreased (P < 0. 01). After genipin was administered, the expression of UCP2 decreased (P < 0. 05), and the expression of antioxidative protein increased (P < 0. 05). Experiments suggest that genipin has anantioxidant protective effect on MIN6 cells damaged by high glucose and maintains the function of MIN6cells to promote insulin secretion. This experiment provides experimental data for the antioxidation of genipin on MIN6 cells injured by high glucose, and also provides new ideas for the follow-up study of genipin in the treatment and prevention of diabetes.
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AIM:To explore the effects of genipin (GEN) on high glucose (HG) -induced oxidative stress injury and apoptosis in H9c2 cardiomyocytes.METHODS:H9c2 cells were cultured in vitro and HG-induced injury model was established.H9c2 cells were divided into 4 groups:normal control (NC) group (glucose at 5.6 mmol/L) , HG group (glucose at 50 mmol/L) , NG+GEN group and HG+GEN group.The concentration of genipin was used at 10μmol/L.The viability of the H9c2 cells was measured by CCK-8 assay.The intracellular malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined by enzyme labeling and WST-1 methods, respectively.The activity of lactate dehydrogenase (LDH) in the cell culture supernatant was detected by microplate method.Fluorescent probe DCF was used to detect intracellular levels of reactive oxygen species (ROS).Nucleosome fragments was measured to evaluate cell apoptosis by ELISA.The intracellular mitochondrial membrane potential was detected by JC-1 method.The protein levels of Mn-SOD, cytochrome C (Cyt C) , Bax and cleaved caspase-3 were determined by Western blot.RESULTS:Compared with HG group, the cell viability in HG+GEN group was increased significantly (P<0.05) , the levels of MDA and LDH were decreased (P<0.05) , SOD activity was increased (P<0.05) , the levels of ROS and nucleosome fragments in HG+GEN group were decreased (P<0.05) , and the mitochondrial membranes potential was notably increased (P<0.05).Compared with NG group, the activation of Mn-SOD was decreased, but the protein levels of Cyt C, Bax and cleaved caspase-3 were increased in HG group (P<0.05).Compared with HG group, the activation of Mn-SOD was increased, and the protein levels of Cyt C, Bax and cleaved caspase-3 were decreased in HG+GEN group (P<0.05).CONCLUSION:Genipin protects HG-induced H9c2 cardiomyocytes against oxidative stress injury and apoptosis.
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AIM:To observe the effects of ginsenoside Rh 1 on the levels of inflammatory factors in serum and bronchoalveolar lavage fluid(BALF),and the pathological changes of the lung tissues in an experimentally induced mouse asthma model.METHODS:Male BALB/c mice(n=40)were divided into 4 groups:normal control group,asthma mo-del group,and low-dose(40 mg· kg-1 · d-1 )and high-dose(80 mg· kg-1 · d-1 )ginsenoside Rh1 groups.The bron-chial asthma mouse model was established by the method of ovalbumin induction and excitation ,and during the excitation period,the mice were daily treated with ginsenoside Rh 1 for 2 weeks.At 24 h after the final dose of ginsenoside Rh 1,the mice were sacrificed.The number of eosinophils(EOS)and the concentrations of interleukin(IL)-4,IL-5 and interferon(IFN)-γin BALF were determined.The levels of IgG and IgE in serum were measured ,and the expression of transforming growth factor(TGF)-β1 and the pathological changes in lung tissues were evaluated.RESULTS:Ginsenoside Rh1 inhibi-ted the increases in the number of EOS and the concentrations of IL-4,IL-5,IFN-γand IgE,reversed the increased ex-pression of TGF-β1,and improved the pathological changes of the lung tissues in asthmatic mice.CONCLUSION:Gin-senoside Rh1 improves the immuno-inflammatory profile and pathological changes in the experimentally induced mouse asth -ma model,implying its potential therapeutic effect on asthma.
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Objective To analyze the effect of bone peptide injection in patients with osteoarthritis of the activity of alkaline phosphatase. Methods A total of 74 patients with osteoarthritis of the bone were selected from March 2015 to May 2017 in our hospital and were randomly divided into two groups,with 37 cases in each group.The control group was treated with glucosamine sulfate, the observation group was treated with ossotide injection.The therapeutic effect and alkaline phosphatase activity were compared between two groups. Results The total effective rate was 97.30% in the observation group, which was significantly higher than that in the control group (70.27%) (P<0.05) .The score of knee function was (79.62±5.80) in the observation group.which was significantly higher than that in the control group (71.06±5.41) (P<0.05).The alkaline phosphatase activity after the treatment of 30d and 45d (0.97±0.12), nkat/L (1.39±0.12) nkat /L was significantly higher than the control group (0.71±0.10), nkat/L, (0.88±0.10) nkat/L, the incidence rate of adverse reaction(5.41 %) was significantly lower than the control group (24.32 %) (P<0.05). Conclusion Bone peptide injection has significant therapeutic effect on bone and joint osteoarthritis, can effectively improve the knee joint function of the patients, improve the patients with alkaline phosphatase activity, and high safety, is worthy of promotion.
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Objective To analyze the effect of bone peptide injection in patients with osteoarthritis of the activity of alkaline phosphatase. Methods A total of 74 patients with osteoarthritis of the bone were selected from March 2015 to May 2017 in our hospital and were randomly divided into two groups,with 37 cases in each group.The control group was treated with glucosamine sulfate, the observation group was treated with ossotide injection.The therapeutic effect and alkaline phosphatase activity were compared between two groups. Results The total effective rate was 97.30% in the observation group, which was significantly higher than that in the control group (70.27%) (P<0.05) .The score of knee function was (79.62±5.80) in the observation group.which was significantly higher than that in the control group (71.06±5.41) (P<0.05).The alkaline phosphatase activity after the treatment of 30d and 45d (0.97±0.12), nkat/L (1.39±0.12) nkat /L was significantly higher than the control group (0.71±0.10), nkat/L, (0.88±0.10) nkat/L, the incidence rate of adverse reaction(5.41 %) was significantly lower than the control group (24.32 %) (P<0.05). Conclusion Bone peptide injection has significant therapeutic effect on bone and joint osteoarthritis, can effectively improve the knee joint function of the patients, improve the patients with alkaline phosphatase activity, and high safety, is worthy of promotion.
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To observe the inhibitive effect of Baicalin against influenza A H1N1 virus infection in epithelial cell line A549, the cell proliferation and cytotoxicity were assayed by MTT, the cell cycle and the apoptosis were analyzed by flowcytometer using PI staining, the morphology of cellular nucleolus was observed by Hoechst 33258 staining and the effects of activation on caspase 3 and caspase 8/9 were also detected by immunofluorescent staining with a fluorescence microscope. The results showed that Baicalin exerted an inhibitive effect on CPE after influenza A H1N1 virus infection. The FACS with PI staining showed that the cell cycle of the infected cell was arrested at S phase, the Baicalin-treated group decreased S phase cell ratio and subG0 phase peak in comparison with the control (P < 0.05) and significantly promoted cell proliferation (# P < 0.05). Hoechst33258 staining suggested that Baicalin protected the cellular nucleolus against the influenza virus-induced apoptosis. Observation under the immunofluorescent microscope suggested that the activities of caspase-8 and caspase-3 were enhanced at 36 h post the influenza virus infection, but 100 microg/mL Baicalin suppressing the activation of caspase-8 and caspase-3 rather than that of caspase-9. In summary, this research confirmed that Baicalin inhibited the influenza A H1N1 virus strain infection in vitro, the drug obviously protected cells from apoptosis damages through regulating cell cycle and suppressed the activation of caspase-8 and caspase-3. The down-regulation was significant and showed a dose-dependent relationship.