摘要
目的探讨趋化因子fractalkine(FKN)对小鼠足细胞炎症因子分泌的影响及机制、对nephrin表达的影响及其在足细胞凋亡中的可能机制。方法培养小鼠足细胞,将细胞分为:①对照组;②rmFKN干预组;③rmFKN+SC-514(抑制剂)组。采用流式细胞术检测各组中足细胞的凋亡率。用蛋白质印迹法(Western blot)检测足细胞nephrin、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、细胞核内核因子-κB(NF-κB)、Bax、Bcl-2及细胞色素C(Cytochrome C,Cyt-c)蛋白表达水平。结果凋亡结果显示,各组细胞培养48 h后,与对照组相比,rmFKN+SC-514组凋亡率差异无统计学意义(P>0.05),rmFKN组凋亡率升高,而rmFKN组与rmFKN+SC-514组相比凋亡率升高,差异均有统计学意义(P<0.05)。Western blot结果显示,与对照组相比,rmFKN组nephrin的表达水平下调,rmFKN组IL-6、TNF-α分泌增多,NF-κB p65、Bax、Cyt-c表达水平上调,Bcl-2表达水平下调,差异均有统计学意义(P<0.05)。与rmFKN组相比,rmFKN+SC-514组IL-6、TNF-α蛋白分泌减少,NF-κB p65表达水平下调,差异有统计学意义(P<0.05)。结论①rmFKN可抑制足细胞nephrin的表达;②rmFKN可以促进足细胞IL-6、TNF-α的产生和分泌,NF-κB的活化可能是其潜在机制;③rmFKN促进足细胞的凋亡,其作用可能通过NF-κB信号通路调节Cyt-c的表达及Bax/Bcl-2平衡有关。
Objective To investigate the effect of fractalkine(FKN) on the secretion of inflammatory factors,the expression of nephrin and the possible mechanism of apoptosis in podocyte.Methods Podocytes were cultured and randomly divided into three groups:control group,rmFKN group and rmFKN+SC-514(Inhibitor of NF-κB p65) group.Podocyte apoptosis was detected by flow cytometry.The expression of nephrin,interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),nuclear kernel factor-κB(NF-κB),Bax,Bcl-2,and Cyt-c were detected by western blot.Results The results of flow cytometry showed that there was no significant difference in the apoptosis rate of rmFKN+SC-514 group compared with the control group(P>0.05),and the apoptosis rate of rmFKN group was higher than that of rmFKN + SC-514 group(P<0.05).Western blot showed that compared with the control group,the expression level of nephrin decreased,the secretion of IL-6 and TNF-α increased,the expression level of NF-κB p65,Bax and Cyt-c increased,and the expression level of Bcl-2 decreased in rmFKN group(P<0.05).Compared with the rmFKN group,the secretion of IL-6,TNF-α and the expression level of NF-κ B p65 were decreased in the rmFKN+SC-514 group(P<0.05).Conclusion ①FKN can inhibit the expression of nephrin;②FKN can promote the production and secretion of IL-6 and TNF-α in potocyte,which may be associated with activation of the NF-κB signaling pathway;③FKN can promote the apoptosis of podocytes,which may be related to the regulation of Cyt-c expression and the balance of Bax/Bcl-2 by activating NF-κB signaling pathway.
作者
马敬雪
黄丹
巩奇明
卢俊玲
尤燕舞
MA Jingxue;HUANG Dan;GONG Qiming;LU Junling;YOU Yanwu(Department of Nephrology,Affiliated Hospital,Youjiang Medical University for Nationalities,Baise 533000,China;Graduate School,Youjiang Medical University for Nationalities,Baise 533000,China)
出处
《右江医学》
2020年第1期17-22,共6页
Chinese Youjiang Medical Journal
基金
国家自然科学基金(81860296)
广西研究生教育创新计划项目(YCSW2019221)
