目的 探究益肾通络方对膜性肾病大鼠肾组织表皮生长因子受体（epidermal growth factor receptor，EGFR）及足细胞和肾小管上皮细胞凋亡的影响。方法 SD大鼠尾iv阳离子牛血清白蛋白（cationic bovine serum albumin，C-BSA）建立膜性肾病大鼠模型，给予益肾通络方和盐酸贝那普利进行干预，检测各组大鼠24 h尿蛋白（urine total protein，UTP）、血清中白蛋白（albumin，ALB）和总胆固醇（total cholesterol，TC）水平；观察各组大鼠肾组织病理变化；采用TUNEL法检测各组大鼠肾组织足细胞和肾小管上皮细胞凋亡情况；采用免疫组化法检测各组大鼠肾组织磷酸化EGFR（p-EGFR）和剪切型半胱氨酸蛋白酶-3（cleaved Caspase-3）的表达情况；采用Western blotting法检测各组大鼠肾组织p-EGFR、Ras相关的C3肉毒素底物1（Ras-related C3 botulinum toxin substrate 1，Rac1）、c-jun、c-fos、FasL和cleaved Caspase-3蛋白表达情况。结果 与对照组比较，模型组大鼠24 h UTP以及血清中TC水平明显升高（P<0.01），ALB水平显著降低（P<0.01）；肾小球基底膜、足细胞结构发生改变；肾组织足细胞和肾小管上皮细胞凋亡明显增多（P<0.01）；肾组织p-EGFR、Rac1、c-jun、c-fos、FasL和cleaved Caspase-3蛋白表达水平显著升高（P<0.01）。与模型组比较，益肾通络方中、高剂量组大鼠24 h UTP以及血清中TC水平明显降低（P<0.01），ALB水平明显升高（P<0.01）；肾组织病理损伤得到改善；肾组织足细胞和肾小管上皮细胞凋亡明显减少（P<0.05、0.01），肾组织p-EGFR、Rac1、c-jun、c-fos、FasL和cleaved Caspase-3蛋白表达水平明显降低（P<0.05、0.01）。结论 益肾通络方可以减轻膜性肾病大鼠肾脏病理损伤，抑制肾组织足细胞和肾小管上皮细胞凋亡，其机制可能与抑制EGFR及下游Rac1/c-jun、c-fos/FasL信号途径有关。

Objective To explore the effect of Yishen Tongluo Recipe (益肾通络方) on epidermal growth factor receptor (EGFR), apoptosis of podocytes and renal tubular epithelial cells in renal tissue of rats with membranous nephropathy. Methods SD rats were tail iv cationic bovine serum albumin (C-BSA) to establish a rat model of membranous nephropathy. Yishen Tongluo Recipe and benazepril hydrochloride were given for intervention. The level of 24 h urine total protein (UTP), levels of albumin (ALB) and total cholesterol (TC) in serum of rats in each group were tested; Pathological changes of kidney tissues in rats of each group were observed; TUNEL method was used to detect podocytes and renal tubular epithelial cell apoptosis in kidneys tissue of rats in each group; Immunohistochemical method was used to detect the expressions of phosphorylated EGFR (p-EGFR) and cleaved Caspase-3 in renal tissues of rats in each group; Western blotting was used to detect expressions of p-EGFR, Ras-related C3 botulinum toxin substrate 1 (Rac1), c-jun, c-fos, FasL and cleaved Caspase-3 protein. Results Compared with control group, 24 h UTP and TC level in serum of rats in model group were significantly increased (P < 0.01), ALB level was significantly reduced (P < 0.01); Glomerular basement membrane and podocyte structure were changed; Apoptosis of podocytes and renal tubular epithelial cells in kidney were significantly increased (P < 0.01); Expressions of p-EGFR, Rac1, c-jun, c-fos, FasL and cleaved Caspase-3 in renal tissue were significantly increased (P < 0.01). Compared with model group, 24 h UTP and TC level in serum of rats in medium- and high-dose Yishen Tongluo Recipe groups were significantly reduced (P < 0.01), and ALB level was significantly increased (P < 0.01); Pathological damage of kidney tissue was improved; Podocytes and renal tubular epithelial cell apoptosis in renal tissue were significantly reduced (P < 0.05, 0.01); Expressions of p-EGFR, Rac1, c-jun, c-fos, FasL and cleaved Caspase-3 in renal tissue were significantly reduced (P < 0.05, 0.01). Conclusion Yishen Tongluo Recipe alleviates pathological damage of kidney in rats with membranous nephropathy, inhibit the apoptosis of renal podocytes and renal tubular epithelial cells, mechanism of which may be related to the inhibition of EGFR and downstream Rac1/c-jun, c-fos/FasL signal pathways.

R285.5

河北省自然科学基金青年项目（H2020423049）；河北省中医药管理局科研计划项目（2021008）