摘要
目的:探讨皮诺敛酸(PLA)对脂多糖(LPS)诱导小胶质细胞BV2炎性反应作用及机制研究。方法:MTT法评价PLA对BV2活力影响;细胞免疫荧光法检测自噬蛋白Beclin1、LC3B表达水平;ELISA法检测BV2上清液中IL-1β、IL-6和TNF-α表达影响;分子模拟技术分析PLA与p38MAPK、p65NF-κB蛋白之间的结合能力;Western blot法检测p38MAPK/p65NF-κB信号通路相关蛋白表达情况。结果:与模型组比较,PLA各剂量组细胞存活率显著升高(P<0.05,P<0.01),细胞内Beclin1、LC3B蛋白表达显著减少(P<0.05,P<0.01);PLA中、高剂量组细胞上清液中TNF-α、IL-6和IL-1β含量显著降低(P<0.05);PLA与p38MAPK、p65NF-κB蛋白对接最佳Ki值分别为502.89μmol/L、19.09mmol/L;除PLA中剂量组细胞内p38MAPK蛋白外,其余3组p38MAPK/p65NF-κB蛋白表达水平显著下降(P<0.05,P<0.01)。结论:PLA对LPS诱导BV2细胞炎症损伤具有保护作用,并抑制BV2细胞的自噬,降低炎症因子表达水平,其机制可能与抑制细胞内p38MAPK/p65NF-κB信号通路相关蛋白表达有关。
Objective: To investigate the effects and mechanism of pinoxidic acid(PLA) on lipopolysaccharide(LPS) induced inflammatory response of microglia BV2. Methods: MTT assay was used to evaluate the effect of PLA on BV2 activity. The expression of autophagy proteins Beclin1 and LC3 B was detected by immunofluorescence assay. The expression of IL-1β, IL-6 and TNF-α in BV2 supernatant was detected by ELISA kit. Molecular simulation technology was used to analyze the binding energy between PLA and p38 MAPK, p65 NF-кB;Western blot was used to detect the expression of p38 MAPK/p65 NF-кB signal pathway related proteins. Results: The cell survival rates of PLA low, medium and high dose groups were significantly increased(P<0.05, P<0.01). The protein expression of Beclin1 and LC3 B in PLA low, medium and high dose groups were significantly decreased(P<0.05, P<0.01). The contents of TNF-α, IL-6 and IL-1β of PLA medium and high dose group were significantly decreased(P<0.05);the optimal Ki values of PLA and p38 MAPK and p65 NF-кB proteins were 502.89μmol/L and 19.09 mmol/L, respectively. The expression levels of p38 MAPK/p65 NF-кB protein in PLA low, medium and high dose groups were decreased significantly(P<0.05, P<0.01), except for p38 MAPK protein in PLA medium does group. Conclusion: PLA can protect BV2 cells from LPS induced inflammatory damage, inhibit the autophagy of BV2 cells, and reduce the expression of inflammatory factors. The mechanism may be related to the inhibition of p38 MAPK/p65 NF-кB signal pathway related protein expression.
作者
阚默
宋凤媛
陈锡俊
兰兴成
王继凤
杨擎
李晶
李娜
林喆
KAN Mo;SONG Feng-yuan;CHEN Xi-jun;LAN Xing-cheng;WANG Ji-feng;YANG Qing;LI Jing;LI Na;LIN Zhe(Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2020年第5期2608-2612,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
吉林省科技发展计划项目(No.201603094YY)
