肾疏宁干预HBV诱导的HK-2细胞转分化及迁徙能力的研究

Study on Shenshuning's intervention of HBV-induced transdifferentiation and migration of HK-2 Cells

关键词:  肾疏宁  乙型肝炎病毒相关性肾炎  转分化  Transwell

Key words:  Shenshuning  hepatitis B virus-associated nephritis  transdifferentiation  Transwell

摘要:

[目的]探讨肾疏宁对乙型肝炎病毒（HBV）质粒诱导HK-2细胞发生转分化的干预作用。[方法]制备小鼠含药血清；培养HK-2细胞并进行HBV质粒转染。将细胞分为：空白血清对照组、空质粒组、HBV组、肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组。运用酶联免疫吸附实验（ELISA）、免疫细胞化学法和Transwell细胞体外迁徙实验检测HK-2细胞中乙型肝炎病毒表面抗原（HBsAg）、乙肝病毒e抗原（HBeAg）、α-平滑肌肌动蛋白（α-SMA）、E-钙黏蛋白（E-cadherin）的表达以及细胞迁徙能力。[结果]ELISA检测：HBV组与空质粒组和空白血清组比较，HBsAg、HBeAg表达上调，差异有统计学意义（P<0.01）。免疫细胞化学检测：与HBV组比较，肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组α-SMA蛋白表达明显下调，差异有统计学意义（P<0.01）。与HBV组比较，肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组有上调E-cadherin蛋白表达的趋势，但差异无统计学意义（P>0.05）。与HBV组比较，肾疏宁低浓度组、肾疏宁中浓度组、肾疏宁高浓度组细胞迁徙能力明显减弱，差异有统计学意义（P<0.01）。[结论]HBV转染HK-2细胞会造成细胞转分化并增强细胞迁徙能力。而肾疏宁含药血清可以减轻由HBV导致的上皮细胞转分化并减弱其细胞迁徙能力。

Abstract:

[Objective] To investigate the effect of Shenshuning on the transdifferentiation of HK-2 cells induced by HBV plasmid.[Method] Mouse drug-containing serum was prepared. HK-2 cells were cultured and transfected with HBV plasmid. The cells were divided into:blank serum control group,empty plasmid group,HBV group,Shenshuining low concentration group,Shenshuinging middle concentration group,and Shenshuing high concentration group. The expression of HBsAg,HBeAg,α-SMA,E-cadherin and cell migration ability in HK-2 cells were detected by enzyme-linked immunosorbent assay (ELISA),immunocytochemistry and Transwell cell migration assay.[Results] ELISA detection:HBsAg and HBeAg expression were up-regulated in HBV group compared with empty plasmid group and blank serum group (P<0.01). Immunocytochemical detection:compared with the HBV group,the expression of α-SMA protein in the low-concentration group of Shenshuening,the concentration of Shenshuening and the high concentration of Shenshuing was significantly down-regulated,and the difference was significant (P<0.01). Compared with the HBV group,the low-concentration group of Shenshuing,the concentration of Shenshuing and the high concentration of Shenshuing had an up-regulation of E-cadherin protein expression,but the difference was not statistically significant (P>0.05). Compared with the HBV group,the migration ability of the kidney-sparing low-concentration group,the Shenshuing-Ning-concentration group,and the Shenshu-ning high-concentration group was significantly weakened,and the difference was significant (P<0.01).[Conclusion] HBV transfection of HK-2 cells will cause cell transdifferentiation and enhance cell migration ability. The drug-containing serum of Shen Shu Ning can alleviate the transdifferentiation of epithelial cells caused by HBV and reduce its cell migration ability.