半枝莲黄酮对β-淀粉样蛋白所致星形胶质细胞损伤的影响

Effect of Scutellaria Barbata Flavonoids on β-Amyloid Protein-induced Injury in Rats Astrocytes

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。

Objective To study the effects of flavonoids from Scutellaria Barbara on 13-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 70.0μg·mL-1 , respectively） of Scutellaria Barbata flavonoids groups. After treatment by 17.5, 35.0 and 70.0 μg·mL-1 of Scutellaria Barbata flavonoids for 24 h, the cells of model control, small, middle and large dose groups were added with A1325_35（final concentration： 100 μmol·L-1） and cultured for another 24 h. MTT eolorimetric 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% and 53.60% 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.