【摘要】 目的探讨半枝莲黄酮(SBF)对β-淀粉样蛋白25-35(Aβ25-35)损伤星形胶质细胞的干预作用。方法培养第3代星形胶质细胞分为空白对照组,模型对照组和半枝莲黄酮小、中、大剂量组。半枝莲黄酮小、中、大剂量组细胞分别给予17.5,35.0和70.0μg·m L-1半枝莲黄酮作用24 h后,模型对照组和半枝莲黄酮小、中、大剂量组细胞再分别加入终浓度Aβ25-35100μmol·L-1继续作用24 h。噻唑蓝(MTT)比色法测定细胞存活率,分光光度法测定培养液中乳酸脱氢酶(LDH)的含量,免疫组织化学方法测定细胞内LDH的合成量,酶联免疫吸附测定(ELISA)法测定培养液中细胞因子白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的水平。结果与空白对照组比较,模型对照组细胞存活率降低49.94%(P<0.01)。与模型对照组比较,半枝莲黄酮小、中和大剂量组细胞存活率分别提高12.50%(P<0.05),16.67%(P<0.01)和2.08%(P>0.05)。与空白对照组比较,模型对照组细胞培养上清液中LDH减少36.65%(P<0.01)。与模型对照组比较,半枝莲黄酮小、中和大剂量组LDH分别增加11.35%(P>0.05),26.40%(P<0.01)和32.44%(P<0.01)。与空白对照组比较,模型对照组星形胶质细胞LDH蛋白表达的阳性面积比例降低24.92%(P<0.05)。与模型对照组比较,半枝莲黄酮中和大剂量组LDH蛋白表达的阳性面积比例分别提高31.20%和53.60%。与空白对照组比较,模型对照组细胞培养液中IL-1β和IL-6水平分别降低23.60%和15.47%(P<0.01)。与模型对照组比较,半枝莲黄酮小、中和大剂量组IL-1β释放量分别增加5.74%(P>0.05),24.82%(P<0.01)和6.86%(P>0.05);IL-6释放量分别增加11.30%(P<0.05),17.39%(P<0.01)和3.87%(P>0.05)。结论半枝莲黄酮对Aβ25-35引起的星形胶质细胞损伤具有保护作用,这可能有利于阐述半枝莲黄酮治疗阿尔茨海默病的作用机制。更多还原

【Abstract】 Objective To study the effects of flavonoids from Scutellaria Barbata on β-amyloid protein 25-35( Aβ25-35)-induced injury of rats’ astrocytes. Methods Rats’ astrocytes of passage 3 were divided into blank control group,model control group,small,middle and large doses( 17. 5,35. 0 and 70. 0 μg · m L- 1,respectively) of Scutellaria Barbata flavonoids groups. After treatment by 17. 5,35. 0 and 70. 0 μg·m L- 1of Scutellaria Barbata flavonoids for 24 h,the cells of model control,small,middle and large dose groups were added with Aβ25-35( final concentration: 100 μmol·L- 1) and cultured for another 24 h. MTT colorimetric method was used to measure cell viability,spectrophotometry to determine LDH content in cultured medium,immunohistochemistry to detect cellular LDH,ELISA to detect IL-1β and IL-6 levels in culture medium.Results As compared with blank control group,cell survival rate of model control group was significantly decreased by 49. 94%( P < 0. 01). As compared with model control group,cell survival rate was increased by 12. 50%( P < 0. 05),16. 67%( P <0. 01) and 2. 08%( P > 0. 05) in small,medium and large dose groups,respectively. As compared with blank control group,LDH of supernatant fluid was significantly decreased by 36. 65%( P < 0. 01). As compared with model control group,LDH level was increased by 11. 35%( P > 0. 05),26. 40%( P < 0. 01) and 32. 44%( P < 0. 01) in the small,medium and large dose groups. As compared with blank control group,positive area proportion of LDH protein in astrocytes was decreased by 24. 92%( P < 0. 05). As compared with model control group,positive area proportion of LDH protein was increased by 31. 20% and53. 60% in medium and large dose groups,respectively. As compared with blank control group,IL-1β and IL-6 were significantly decreased in the model control group by 23. 60% and 15. 47%( P < 0. 01),respectively. As compared with model control group,IL-1β and IL-6 were increased in small,medium and large dose groups,and IL-1β was increased by 5. 74%( P > 0. 05),24. 82%( P < 0. 01) and 6. 86%( P > 0. 05),respectively; IL-6 was increased by 11. 30%( P < 0. 05),17. 39%( P < 0. 01)and 3. 87%( P > 0. 05),respectively. Conclusion Scutellaria Barbata falvonoids exert protective effect on Aβ25-35-induced astrocyte injury. Scutellaria Barbata flavonoids might treat Alzheimer’s disease through influencing astrocytes.更多还原