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目的以原代培养的大鼠大脑星形胶质细胞进行缺氧缺糖再给氧处理,观察党参总皂苷对星形胶质细胞损伤的保护作用。方法1 d龄W istar大鼠乳鼠大脑星形胶质细胞原代培养,含连二亚硫酸钠的无糖Earle’s液进行缺氧缺糖处理造模,MTT法测定细胞存活率,Hoechst 33342和PI原位双染法荧光显微镜观察细胞形态学变化和细胞坏死、凋亡率,比色法测定LDH漏出率,评价药效。结果与模型组比较,党参总皂苷Ⅰ、Ⅲ组均能显著提高细胞存活率(P<0.01),党参总皂苷Ⅰ、Ⅱ、Ⅲ组均能显著降低细胞坏死率(P<0.01),党参总皂苷各组对细胞凋亡率均无显著影响(P>0.05),但均能显著降低细胞LDH漏出率(P<0.01)。结论党参总皂苷对缺血再灌注损伤后星形胶质细胞的坏死具有显著抑制作用,但对凋亡过程无保护作用,提示党参总皂苷是党参治疗中风病急性期的主要效应成分。
Objective To observe the protective effects of Codonopsis pilosula total saponins on astrocytes after primary cultured rat astrocytes were treated with hypoxia and glucose deprivation. METHODS: One-day-old W istar rat neonatal brain astrocytes were cultured in primary culture. Sugar-free Earle’s solution containing sodium dithionite was used to model hypoxia-glucose deprivation. MTT assay was used to determine cell viability, Hoechst 33342 and PI were used. Double-staining fluorescence microscopy was used to observe the cell morphological changes, cell necrosis and apoptosis rate. The colorimetric method was used to determine the leakage rate of LDH and evaluate the efficacy. Results Compared with the model group, Codonopsis saponins I and III groups could significantly increase the cell survival rate (P<0.01). Codonopsis saponins I, II, and III groups could significantly reduce the rate of cell necrosis (P<0.01). Saponin had no significant effect on the cell apoptosis rate (P>0.05), but all of them could significantly reduce the cell LDH leakage rate (P<0.01). Conclusion The total saponins of Radix Codonopsis can significantly inhibit the necrosis of astrocytes after ischemia-reperfusion injury, but it has no protective effect on the apoptosis process, suggesting that the total saponins of Radix Codonopsis is the main effect component of Radix Codonopsitis in the treatment of apoplexy.