目的 研究三焦祛湿方对阳离子化牛血清白蛋白（cationic bovine serum albumin，C-BSA）诱导的膜性肾病（membranous nephropathy，MN）小鼠的作用及其对烟酰胺腺嘌呤二核苷酸磷酸氧化酶4（nicotinamide adenine dinucleotide phosphate oxidase 4，NOX4）/活性氧（reactive oxygen species，ROS）/p38信号通路的影响，以阐明其抗氧化应激的作用机制。方法 将60只小鼠随机分为对照组10只和造模组50只，造模组尾iv 6.5 mg/kg C-BSA建立MN小鼠模型，随机分为模型组及三焦祛湿方低、高剂量（3.71、7.42 g/kg）组和盐酸贝那普利（1.3 mg/kg）组，各给药组ig相应药物干预4周，检测各组小鼠24 h尿蛋白；腹主动脉取血，检测血清丙二醛（malondialdehyde，MDA）水平和超氧化物歧化酶（superoxide dismutase，SOD）活性；取肾组织进行苏木素-伊红（HE）、Masson、过碘酸-六胺银（PASM）染色及透射电镜（TEM），观察肾组织病理形态变化，采用免疫荧光法检测肾组织免疫球蛋白G（immunoglobulin G，IgG）沉积和ROS表达情况；采用Western blotting法检测肾组织NOX4、p38和磷酸化p38（p-p38）蛋白表达情况。结果 与对照组相比，模型组小鼠24 h尿蛋白显著升高（P<0.01）；血清SOD活性显著降低，MDA水平显著升高（P<0.01）；肾脏病理改变明显，可见肾小球体积增大，基底膜增厚，嗜复红蛋白沉积，“钉突”形成，上皮下可见电子致密物沉积，足突融合，IgG沿毛细血管襻颗粒状沉积；肾组织ROS表达显著增多（P<0.01），NOX4、p-p38蛋白表达水平显著升高（P<0.01）。与模型组相比，各给药组小鼠24 h尿蛋白显著降低（P<0.01）；血清SOD活性明显增强，MDA水平显著降低（P<0.05、0.01）；肾组织病理损害明显改善；肾组织ROS表达显著减少（P<0.01），NOX4和p-p38蛋白表达水平显著降低（P<0.05、0.01）。结论 三焦祛湿方能够减少尿蛋白，减轻肾脏病理损伤，缓解肾组织氧化应激，其作用机制可能与调控NOX4/ROS/p38信号通路有关。

Objective To study the effects of Sanjiao Qushi Prescription (三焦祛湿方) on cationic bovine serum albumin (C-BSA) induced membranous nephropathy (MN) mice model and nicotinamide adenine dinucleotide phosphatase oxidase4 (NOX4)/reactive oxygen species (ROS)/p38 signaling pathway, and elucidate the mechanism of anti-oxidative stress.Methods Sixty mice were randomly divided into control group (n=10) and model group (n=50), mice were iv C-BSA (6.5 mg/kg) by tail to establish MN model,

which were randomly divided into model group, low-, high-dose Sanjiao Qushi Prescription (3.71, 7.42 g/kg) groups and benazepril hydrochloride (1.3 mg/kg) group. Mice were ig corresponding drugs for four weeks, 24 h urine protein were detected. Serum was collected, superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were measured. Renal tissue was collected, pathological morphology changes were observed by light microscope with hematoxylineosin (HE), Masson and periodic acid-silver metheramine (PASM) staining and transmission electron microscope (TEM). Deposition of immunoglobulin G (IgG) and ROS expression in renal tissue was observed by fluorescence immunoassay. Western blotting was used to detect NOX4, p38 and p-p38 protein expressions in renal tissue. Results Compared with control group, 24 h urine protein of mice in model group was significantly increased (P < 0.01), SOD activity was decreased and MDA level was increased (P < 0.01) in model group; Kidney exhibited enlarged volume of glomerular, significantly thickened glomerular basement membrane, polymyoglobin deposition and spike formation from the light microscope, epithelium deposition of electron dense, foot process fusion were observed from transmission electron microscope; Granular deposition of IgG along the capillary wall was observed by immunofluorescence staining; ROS expression in kidney was significantly increased (P < 0.01); NOX4 and p-p38 protein expressions were significantly increased (P < 0.01). Compared with model group, 24 h urine protein in each administration group was significantly decreased (P < 0.01), SOD activity was obviously increased and MDA level was decreased (P < 0.05, 0.01); pathological damages of renal were alleviated; ROS expression in renal tissue was significantly decreased (P < 0.01); NOX4 and p-p38 protein expressions in renal tissue were significantly decreased (P < 0.05, 0.01).Conclusion Sanjiao Qushi Prescription can decrease 24 h urine protein, ameliorate renal pathological damages, alleviate oxidative stress in kidney, its mechanism may be possibly related to the regulation of NOX4/ROS/p38 signaling pathway.

R285.5

河北省政府资助省级临床医学优秀人才项目（2016034829）；河北省中医药管理局科研计划项目（2021039）；河北省研究生创新资助项目（XCXZZBS2021021）