摘要
目的:研究西洋参茎叶总皂苷(PQS)对氧糖剥夺/复氧(OGD/R)引起的小鼠BV-2小胶质细胞炎性活化的抑制作用,并初步阐明其作用机制。方法:BV-2细胞分为对照组、OGD/R组、PQS各剂量+OGD/R组。对照组细胞在常规条件下培养;OGD/R组细胞在OGD/R条件下培养;PQS各剂量+OGD/R组细胞给予PQS(30、60、90μg/mL)预处理后,转入OGD/R条件培养。通过酶联免疫法检测上清液中炎性因子肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的含量,通过CCK-8法检测PQS对OGD/R条件下细胞增殖活性的影响,流式细胞术检测胶质细胞活化标志蛋白离子钙结合适配器分子1(Iba-1)和酸性溶霉菌糖蛋白(CD68)的表达,Western blot检测核因子-κB(NF-κB)p65磷酸化及其抑制因子NF-κB抑制蛋白α(IκB-α)和NF-κB抑制因子(NKRF)蛋白的表达。结果:OGD/R诱导能够明显增加BV-2细胞上清液中炎性因子TNF-α、IL-1β和IL-6含量(P<0.01),增强BV-2细胞对数期增殖活性(P<0.01),上调CD68表达和p-NF-κB p65(P<0.01),同时下调Iba-1和NKRF表达(P<0.01)。PQS预处理能够有效逆转BV-2细胞中OGD/R引起的上述指标变化趋势,且其作用呈浓度依赖性(P<0.05,P<0.01)。此外,各组BV-2细胞中IκB-α蛋白表达无明显差异(P>0.05)。结论:PQS能够有效抑制OGD/R引起的BV-2细胞炎性活化,其作用机制可能与上调NKRF蛋白表达有关。
Objective:To study the inhibitory effect of Panax quinquefolium saponin from stems and leaves(PQS)on the inflammatory activation of mouse BV-2 microglia induced by oxygen-glucose deprivation/reoxygenation(OGD/R),and to elucidate its mechanism.Methods:BV-2 cells were divided into the control group,OGD/R group and PQS low,medium,high dose+OGD/R group.The cells in control group were cultured under conventional condition;the cells in OCD/R group were cultured under OGD/R condition;the cells in PQS low,medium,high dose+OCD/R group were pretreated with PQS(30,60,90μg/mL),then cultured in OGD/R condition.The cells were cultured for 48 h,then the contents of inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 in the cell supernatant were detected by ELISA.The proliferative activity was detected by CCK-8 method,the expression of glial activation marker protein ion calcium-binding adapter molecule 1(Iba-1)and acid lysozyme glycoprotein(CD68)were detected by flow cytometry,and the phosphorylation of nuclear factor-KB(NF-kB)p65 and its inhibitory factors NF-kB inhibitory proteinα(IBK-α)as well as NF-kB inhibitory factor(NKRF)were detected by Western blot.Results:OCD/R induction could significantly increase the contents of inflammatory factors such as TNF-a,IL-1βand IL-6 in the supernatant of BV-2 cells(P<0.01),enhance the proliferation activity of BV-2 cells(P<0.01),up-regulate the expression of CD68 and the phosphorylation of NF-kB-p65(P<0.01),and down-regulate the expression of Iba-1 and NKRF(P<0.01).PQS pretreatment could effectively reverse the change trend of the above indexes caused by OGD/R in BV-2 cells,and the effects were concentration-gradient dependent(P<0.05,P<0.01).In addition,there was no significant difference in the expression of IkB-αprotein in BV-2 cells among all groups(P>0.05).Conclusion:PQS can inhibit the inflammatory activation of BV-2 cells induced by OGD/R,and its mechanism may be related to the up-regulation of NKRF protein expression.
作者
刘静
陈希
金宏飞
张英辉
LIU Jing;CHEN Xi;JIN Hongfei;ZHANG Yinghui(Department of Laboratory Medicine,General Hospital of Northern Theater Command,Shenyang 110016,Liaoning,China)
出处
《上海中医药大学学报》
CAS
2023年第2期7-13,共7页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
辽宁省自然科学基金资助项目(2022-MS-046)。
