肾苏Ⅱ方对局灶节段性肾小球硬化大鼠Notch-PI3K/PKB通路串话及足细胞EMT的影响

Effects of Shensu Ⅱ Formula on Notch-PI3K/PKB signaling crosstalk and podocyte EMT in rats with focal segmental glomerular sclerosis

目的:观察中药肾苏Ⅱ方对局灶节段性肾小球硬化(FSGS)大鼠肾脏功能、病理、足细胞及上皮细胞向间充质细胞转分化(EMT)标志蛋白、Notch、磷脂酰肌醇3激酶/蛋白激酶B(PI3K/PKB)通路及其串话关键位点的影响。方法:采用左肾切除+阿霉素注射法诱发SD大鼠FSGS模型,依体质量随机分为3组,模型组、贝那普利组和肾苏Ⅱ方组。另设空白对照组,每组各12只。贝那普利组予贝那普利(9.25mg·kg-1·d-1)灌胃、肾苏Ⅱ方组予肾苏Ⅱ方(9.71g·kg-1·d-1)灌胃,空白对照、模型组予等量0.9%氯化钠溶液灌胃。检测大鼠24h尿微量蛋白(24h Upro)、血清总蛋白(TP)、血清白蛋白(ALB)。观察肾脏病理形态学改变,分析足细胞标志CD2相关蛋白(CD2AP)及EMT标志desmin蛋白表达,Notch、PI3K/PKB通路关键位点的notch1、Hes-1、染色体10上缺失的磷酸酶与张力蛋白同源物基因(PTEN)、PKB基因及p-PKB蛋白表达。结果:与模型组相比,治疗第8周末开始,贝那普利组及肾苏Ⅱ方组FSGS大鼠24h Upro、TP、ALB均明显改善(P<0.05);至治疗第12周末,贝那普利组及肾苏Ⅱ方组大鼠光镜、电镜肾脏病理改变明显减轻;CD2AP蛋白表达恢复与desmin蛋白表达减少明显(P<0.05);Notch1、Hes-1、PTEN m RNA及p-PKB蛋白表达与模型组比较,差异也有统计学意义(P<0.05)。结论:肾苏Ⅱ方可能通过调控Notch-PI3K/PKB通路串话,阻抑足细胞EMT进程,进而延缓FSGS大鼠疾病进展。

Objective： To observe the effects of Shensu Ⅱ Formula on the renal function, pathologic change, marker proteins of podocyte epithelial-mesenchymal transition（EMT）, key positions of Notch-PI3K/PKB pathways crosstalk in rats with focal segmental glomerular sclerosis（FSGS）. Methods： SD rat model of FSGS was established by Adriamycin Injection ＋ left nephrectomy. The rats were randomly divided into 3 groups： the model group（B group）, the benazepril group（C group）, and Shensu Ⅱ Formula group（D group） according to body weight. Besides, another 12 rats were taken as the control group（A group）. Then benazepril（9.25mg·kg-1·d-1） was given to rats in C group and Shensu Ⅱ（9.71g·kg-1·d-1） was given to rats in D group by gavage, while normal saline was given to rats in other groups. The changes of 24-hour urine protein（24h Upro）, total protein（TP）, albumin（ALB） were detected. The renal pathological morphological changes were observed. Expressions of CD2-associated protein（CD2AP） and desmin, m RNA expression of Notch1, Hes-1, phosphase and tensin homology deleted on chromosome ten（PTEN） and PKB, protein expression of p-PKB were determined. Results： At the end of the 8th week, significant difference of 24 h Upro, TP and ALB were shown in C and D groups compared with B group（P〈0.05）. At the end of the 12 th week, renal pathologic changes with light microscope and electron microscope were significantly reduced, expression recovery of CD2 AP and expression of desmin were significantly decreased（P〈0.05）, expressions of Notch 1, Hes-1, PTEN m RNA and p-PKB proteinwere significant differences compared with B group（P〈0.05）. Conclusion： Shensu Ⅱ Formula could postpone the development of renal fibrosis in rats with FSGS through regulation of Notch-PI3K/PKB signaling crosstalk and the inhibition of podocyte EMT process.