摘要
目的通过对β-淀粉样蛋白1-42(Aβ_(1-42))诱导的人神经母细胞瘤细胞(SH-SY5Y细胞)损伤模型的研究,以Caspase-1/GSDMD-N为切入点,探讨六味地黄丸含药血清对SH-SY5Y细胞焦亡的影响。方法以终浓度为20μmol·L^(-1)Aβ_(1-42)诱导SH-SY5Y细胞建立细胞损伤模型。实验分为正常对照组、模型组和六味地黄丸含药血清组。采用台盼蓝染色法观察各组细胞形态并计算细胞成活率;采用酶联免疫吸附法检测细胞上清液中的白细胞介素1β(IL-1β)和白细胞介素18(IL-18)的含量。采用蛋白质印迹法检测细胞中Caspase-1和GSDMD-N的蛋白表达。结果与正常对照组相比,模型组细胞成活率显著降低(P<0.01),上清液中IL-1β和IL-18的含量显著增多(P<0.01),Caspase-1和GSDMD-N的蛋白表达显著升高(P<0.01);与模型组相比,含药血清组细胞成活率显著增高(P<0.01),上清液中IL-1β和IL-18的含量显著减少(P<0.01),Caspase-1和GSDMD-N的蛋白表达显著降低(P<0.01)。结论六味地黄丸含药血清可通过调节Caspase-1/GSDMD-N通路抑制细胞免疫炎性反应,进而抑制细胞焦亡,从而对损伤的SH-SY5Y细胞有保护作用。
Objective To determine the injury model of human neuroblastoma cells(SH-SY5Y)induced by amyliod-beta protein 1-42(Aβ1-42),and to determine the effect of Liuwei Dihuang pills containing serum on SH-SY5Y cell death via Caspase-1/GSDMD-N pathway.Methods SH-SY5Y cell were induced by 20μmol·L^(-1)Aβ_(1-42).The experiment was divided into a normal control group,a model group and a Liuwei Dihuang pills group.Trypan blue staining was used to observe the cell morphology and calculate the cell survival rate.Enzyme-linked immunosorbent assay(ELISA)was used to detect the content of interleukin-1β(IL-1β)and interleukin-18(IL-18)in the cell supernatant.The protein expression of Caspase-1/GSDMD-N was detected by Western blot.Results Compared with the normal control group,the cell survival rate in the model group was lower(P<0.01),the content of IL-1βand IL-18 in the supernatant was increased(P<0.01),and the protein expression of Caspase-1 and GSDMD-N were significantly increased(P<0.01).Compared with the model group,the cell survival rate in the serum containing group was increased(P<0.01),the contents of IL-1βand IL-18 in the supernatant was decreased(P<0.01),and the protein expression of Caspase-1 and GSDMD-N was decreased(P<0.01).Conclusion Liuwei Dihuang pills containing serum can inhibit the cellular immune inflammatory responses by regulating the Caspase-1/GSDMD-N pathway,and further inhibit cell apoptosis,thus protecting SH-SY5Y cells from injury.
作者
高琳琳
赵丹玉
冯晓帆
张林
朱仲康
张欣
柳春
GAO Lin-lin;ZHAO Dan-yu;FENG Xiao-fan;ZHANG Lin;ZHU Zhong-kang;ZHANG Xin;LIU Chun(Liaoning University of Traditional Chinese Medicine,Shenyang 110847)
出处
《中南药学》
CAS
2021年第3期450-454,共5页
Central South Pharmacy
基金
辽宁省自然科学基金项目(No.20180551041)。
