Effects Of Geniposide On Astrocytes In Mice With Experimental Autoimmune Encephalomyelitis

Objective: The effects of geniposide on the release of tumor necrosis factor-?(TNF-?)and interleukin-6(IL-6)by lipopolysaccharide-induced astrocytes were studied by isolating and culturing astrocytes from mouse cerebral cortex in vitro,and the molecular mechanism of anti-inflammatory response and immunity induced by Geniposide in astrocytes of experimental autoimmune encephalomyelitis mice was explored.Methods: 1.ICR newborn mice born 1-2 days were selected to isolate the cerebral cortex of mice.Primary astrocytes were purified and cultured.The cells were identified by morphology and immunofluorescence with glial fibrillary acidic protein.2.By means of morphological observation and CCK-8 experiment,we determined that LPS acted on astrocytes at different concentrations and at different times,and simulated and manufactured a qualified central inflammatory environment model.3.By morphological observation and CCK-8 test,the toxicity and activity of gardenoside on astrocyte were detected,and the optimum concentration and time of action were determined.4.Enzyme-linked immunosorbent assay(ELISA)was used to determine the effect of geniposide on the secretion of inflammatory factors TNF-alpha and IL-6 by astrocytes activated by LPS.5.The experimental results were analyzed by SPSS17.0.Results: 1.After culture and identification,the purity of primary astrocyte is above95%,which meets the requirements of the next experiment.2.Lipopolysaccharide stimulated astrocyte,the number of cells increased,cell body hypertrophy,processes increased.3.Compared with the control group,the levels of IL-6 and TNF-a in the supernatant of LPS group increased significantly(P < 0.01),and increased in a dose-dependent manner over time.4.Lipopolysaccharide can enhance cell proliferation activity,and the effect is best when the concentration is 1 g/ml and the duration is 24 hours.5.When gardenoside concentration was 500 mol/L,1000 mol/L and 2000 mol/L,the activity of astrocytes decreased and cytotoxicity was produced.6.Geniposide had no effect on the proliferation of astrocytes in the range of 0.5-100 mol/L.7.The concentration ofGeniposide below 100 mol/L had no significant difference compared with the control group at different incubation time of 3 h,6 h,12 h,24 h,48 h and 72 h(P > 0.05),indicating that the concentration of Geniposide below 100 mol/L had no effect on the activity of astrocyte and had no cytotoxicity.8.Compared with model group,1,10 and 100 mol/L geniposide could significantly reduce the levels of TNF-a and IL-6 inflammatory factors(P < 0.01),especially IL-6.9.100 mol/L geniposide could inhibit the secretion of IL-6 and TNF-alpha at 24 hours.Conclusion:1.Lipopolysaccharide(1 g/ml,24h)can significantly enhance cell proliferation activity and increase the release of inflammatory factors.2.Geniposide can inhibit the activity of astrocytes and reduce the release of inflammatory factors IL-6 and TNF-alpha.