ABSTRACT
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.
ABSTRACT
Objective: To investigate the effects of long-chain noncoding RNA Linc00673 overexpression on proliferation and apoptosis of gastric cancer cells and its mechanisms. Methods: The recombinant lentivirus expressing plasmid pLVX-Linc00673 and the control empty plasmid pLVX-NC were packaged and amplified in 293T cells, and the recombinant lentivirus was transfected into gastric cancer cell line MGC-803 to establish a cell line stably overexpressing Linc00673. The expression of Linc00673 gene was detected by real-time fluorescence quantitative PCR. The growth and proliferation of cells were observed by MTT assay and clone formation assay. Cell cycle and apoptosis were detected by flow cytometry. The expressions of cell cycle related regulatory genes were detected by qPCR. The expressions of key molecules in the PI3K/Akt signaling pathway and tumor proliferation related proteins were detected by Western blot. Results: The expressions of Linc00673 in gastric cancer cell line MGC-803, BGC-823 and AGS were significantly higher than that in normal gastric mucosa cell line GES-1 (P＜0.05). MGC-803 cell line with stable overexpression of LINC00673 was established, and the expression level of LincC00673 was 200 times higher than that of the control empty carrier group. Overexpression of Linc00673 promoted proliferation of MGC-803 cells (P＜0.05) and clone formation (P＜0.05), inhibited cell apoptosis and affected the G1→S phase progression of cell cycle (P＜0.01). Overexpression of Linc00673 could affect the expressions of cell cycle regulatory gene CCNG2, P19 and CDK1 in MGC-803. Western blot showed that Linc00673 overexpression not only promoted the expressions of the key molecule pAkt in PI3K / Akt signaling pathway and its downstream target NF-κ B and Bcl-2 protein, but also up regulated the expressions of tumor related factors β-catenin and EZH2 proteins. Conclusion: Overexpression of Linc00673 may promote proliferation and inhibit apoptosis of MGC-803 cells through PI3K/Akt signaling pathway.
Subject(s)
Humans , Apoptosis , Cell Line, Tumor , Cell Proliferation , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt/metabolism , RNA, Long Noncoding/genetics , Stomach Neoplasms/pathologyABSTRACT
Long-term regular ethanol intake could change the gut microbiota and affect anxiety and depression-like behaviors․ It is not clear that whether ethanol withdrawal after short-term low-dose drinking has an effect on the gut microbiota or whether it is related to anxiety-like behaviors․ In this study, 30 male Sprague￣Dawley rats were randomly divided into three groups: Ethanol-C: ethanol treatment group, treated with (5 g / kg, 25% V / V) ethanol for 14 days; Ethanol-2: ethanol withdrawal group, treated with (5 g / kg, 25% V / V) ethanol for 14 days and then withdrawal for one day; Ethanol-0: Control group, the rats were given the same amount of distilled water for 14 days․ Feces were collected from all rats, and high-throughput sequencing methods were used to analyze the effect of ethanol withdrawal after short-term low-dose drinking on the gut microbiota․ The open-field test and elevated plus-maze test were used to determine anxiety-like behaviors, and analyze the correlation between gut microbes and anxiety-like behaviors caused by alcohol withdrawal․ Taxonomic analysis of gut microbiota found that the composition and abundance in the ethanol withdrawal group were significantly different from those in the control group and the alcohol-treated group․ The Alpha diversity of gut microbiota in the ethanol withdrawal group was not significantly different from the control group and the ethanol treatment group, whereas the microbial community structure was significantly different․ The percentage of time spent in the open arms and total distance of rats in the ethanol withdrawal group were significantly reduced (P < 0․ 05) , and behavioral parameters were significantly positive correlated with Bacteroides, Fusobacterium, and Escherichia-Shigella (P < 0․ 05) , but significantly negative correlated with Rumenococcus, Trichospirillum and other bacteria (P < 0․ 05) ․ This study suggests that short-term low-dose ethanol withdrawal does not affect the Alpha diversity, but can change the abundance and community structure of the gut microbiota in rats, and the gut microbiota are correlated with anxiety-like behaviors in rats․ This study clarified the changes of gut microbiota after short-term low-dose ethanol withdrawal, and provided a new direction for the study of anxiety-like behaviors caused by short-term low-dose ethanol withdrawal․
ABSTRACT
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.