参芪蛭龙汤含药血清对PAN损伤MPC5细胞凋亡的抑制作用  

Effect of Serum Containing Shenqi Zhulong Decoction(参芪蛭龙汤)on Apoptosis of MPC5 Cells Damaged by PAN

作　　者：薛丕良 王文文 李星宇 刘星 王顺 XUE Peiliang;ZHUANG Yan;LI Xingyu;LIU Xing;WANG Shun(Heilongjiang Institute of Traditional Chinese Medicine,Harbin 150036,Heilongjiang,China;Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036 [2]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《辽宁中医杂志》2024年第4期196-199,I0004,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：黑龙江省自然科学基金项目(LH2020H104)。

摘　　要：目的探讨参芪蛭龙汤含药血清对puromycin aminonucleoside(PAN)损伤MPC5细胞凋亡的抑制作用。方法制备参芪蛭龙汤大鼠含药血清。MPC5细胞随机分为空白组、模型组、给药组,空白组和模型组用10%大鼠空白血清干预,给药组用10%参芪蛭龙汤含药血清干预,培养24 h后,模型组和给药组加入50 mg/L的PAN干预24 h后,倒置显微镜下观察MPC5细胞的形态结构,MTS法检测细胞增殖,Hoechst33342/PI染色法检测细胞凋亡,AnnexinV-FITC/PI双染法检测细胞凋亡,Western-blot法检测MPC5细胞中BAX、Bcl-2、Caspase-3、Nephrin、Podocin蛋白的表达。结果与模型组比较,足突结构较明显,数量较多;能够促进PAN处理后MPC5细胞的增殖(P<0.05);降低细胞凋亡率(P<0.05,P<0.01);降低细胞中Bax蛋白的表达(P<0.01)、降低cleaved-Caspase-3/Caspase-3蛋白比值(P<0.05),升高Bcl-2、Nephrin蛋白的表达(P<0.05,P<0.01)。结论参芪蛭龙汤能够修复裂孔隔膜蛋白、减少足细胞凋亡,对维持肾小球的正常滤过功能具有改善作用,从而减少蛋白尿。Objective To investigate the effect of serum containing Shenqi Zhulong Decoction(参芪蛭龙汤)on apoptosis of MPC5 cells damaged by puromycin aminonucleoside(PAN).Methods Serum containing Shenqi Zhulong Decoction was prepared.MPC5 cells were randomly divided into blank group,model group and drug administration group.The blank group and model group were treated with 10%blank serum,and the drug administration group was treated with 10%serum containing Shenqi Zhulong Decoction.After 24 h of culture,the model group and drug administration group were treated with 50 mg/L PAN for 24 h.The morphological structure of MPC5 cells was observed under inverted microscope.The cell proliferation was detected by MTS.The apoptosis was detected by Hoechst33342/PI staining.The apoptosis was detected by AnnexinV-FITC/PI double staining.The expressions of BAX,Bcl-2,Caspase-3,nephrin and podocin in MPC5 cells were detected by Western-blot.Results Compared with those of the model group,the structure of the foot process was more obvious and the number was more.It could promote the proliferation of MPC5 cells after PAN treatment(P<0.05).The apoptosis rate decreased(P<0.05,P<0.01).The expression of Bax protein was decreased(P<0.01),the cleaved Caspase-3/Caspase-3 ratio was decreased(P<0.05)and the expressions of Bcl-2 and Nephrin protein were increased(P<0.05,P<0.01).Conclusion Shenqi Zhulong Decoction can repair the hiatus membrane protein,reduce the apoptosis of podocytes and improve the normal filtration function of glomeruli,so as to reduce albuminuria.

关 键 词：膜性肾病 足细胞 PAN 细胞凋亡 裂孔隔膜 

分 类 号：R285.5[医药卫生—中药学]