基于PI3K/Akt/NF-κB信号通路探讨加味三仁汤对脂多糖诱导的大鼠肾小球系膜细胞增殖的影响  

Effect of Modified Sanrentang on Lipopolysaccharide-induced Proliferation of Rat Glomerular Mesangial Cells Based on PI3K/Akt/NF-κB Pathway

作　　者：王广伟 张禹[1] 张佳宁 郭登洲 WANG Guang-wei;ZHANG Yu;ZHANG Jia-ning;GUO Deng-zhou(Hebei University of Traditional Chinese Medicine(TCM),Shijiazhuang 050200,China;Shijiazhuang Hospital of TCM,Shijiazhuang 050051,China;Hebei Women's Vocational College,Shijiazhuang 050093,China;The First Affiliated Hospital of Hebei University of TCM,Shijiazhuang 050011,China)

机构地区：[1]河北中医学院,石家庄050200 [2]石家庄市中医院,石家庄050051 [3]河北女子职业技术学院,石家庄050093 [4]河北中医学院第一附属医院,石家庄050011

出　　处：《中国实验方剂学杂志》2022年第5期32-37,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省科技支撑计划项目(16277765D)。

摘　　要：目的:观察加味三仁汤对脂多糖诱导的大鼠肾小球系膜细胞增殖、细胞外基质及磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/核转录因子-κB(NF-κB)信号通路的干预作用,探讨其改善免疫球蛋白A肾病(IgAN)大鼠炎症病变的相关机制。方法:采用血清药理学方法,将18只大鼠分为3组,正常组,加味三仁汤高、低剂量(20.70,10.35 g·kg^(-1)·d^(-1))组,每组6只,加味三仁汤高、低剂量组给予加味三仁汤相应溶液灌胃,正常组灌胃等体积蒸馏水。分别灌胃5 d后,采血制取含药血清。采用大鼠肾小球系膜细胞(HBZY-1),分为正常组,模型组,贝那普利组(50μmol·L^(-1)),加味三仁汤高、低剂量组。采用噻唑蓝(MTT)比色法检测各组细胞增殖情况,酶联免疫吸附测定法(ELISA)检测各组细胞培养上清液中Ⅳ型胶原(ColⅣ)的含量,蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)分别检测PI3K/Akt/NF-κB通路相关蛋白表达量和mRNA表达水平。结果:与正常组比较,模型组大鼠肾小球系膜细胞增殖显著(P<0.01),ColⅣ分泌显著增多(P<0.01),磷酸化(p)-Akt,p-p65蛋白表达显著增加(P<0.01);与模型组比较,加味三仁汤高剂量组显著抑制细胞增殖(P<0.01)和ColⅣ分泌(P<0.01),降低Akt磷酸化水平(P<0.01),降低NF-κB p65阳性表达(P<0.01)。结论:加味三仁汤能够抑制脂多糖诱导的HBZY-1细胞增殖,降低ColⅣ蛋白含量,可能与其抑制PI3K/Akt/NF-κB信号通路激活密切相关。Objective: To observe the intervention of modified Sanrentang on the lipopolysaccharideinduced proliferation of rat glomerular mesangial cells,the phosphatidylinositol-3 kinase(PI3K)/protein kinase B(PKB/Akt)/nuclear factor kappa B(NF-κB)signaling pathway,and to investigate its mechanism in improving kidney inflammation in rats with immunoglobulin A nephropathy(IgAN). Method: The 18 rats were divided into 3 groups by serum pharmacology method:normal group,high-dose and low-dose(20.70,10.35 g·kg^(-1)·d^(-1))groups with 6 rats in each group. Modified Sanrentang high-and low-dose groups were intragastric with the corresponding solution of modified Sanrentang,and normal group was intragastric with equal volume of distilled water. After 5 days of intragastric administration,blood samples were collected to prepare drug-containing serum. Rat mesangial(HBZY-1)were divided into five groups of normal group,LPS 10 mg·L^(-1)in the model group,benazepril(50 μmol·L^(-1)),modified Sanrentang high-and low-dose group. Preclude the use of methyl thiazolyl tetrazolium(MTT) method detect the proliferation activity of HBZY-1 cells, enzyme-linked immunosorbent assay(ELISA)was used to determine the content of each group type Ⅳ collagen(Col Ⅳ),Western blot and Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)were used to detect protein and mRNA expression levels of PI3K/Akt/NF-κB signaling pathway. Result: As compared with the normal group,MTT assay showed that exposure to LPS significantly enhanced the proliferative activity,the Col Ⅳ was increased significantly of HBZY-1 cells(P<0.01),p-Akt,p-p65 was increased significantly(P<0.01). Compared with the model group,the proliferation and Col Ⅳ of rat chronic glomerulonephritis cells induced by LPS by inhibiting PI3K/Akt/NF-κB signaling pathway(P<0.01),and the phosphorylation of Akt was significantly inhibited(P<0.01),the expression levels of NF-κB p65 was reduced in modified Sanrentang highdose group(P<0.01). Conclusion: Modified Sanrentang coul

关 键 词：脂多糖(LPS) 加味三仁汤 磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/核转录因子-κB(NF-κB)信号通路 肾小球系膜细胞(HBZY-1) Ⅳ型胶原(ColⅣ) 

分 类 号：R2-0[医药卫生—中医学] R285