基于醛固酮诱导的ERK/NF-κB信号通路探讨益肾散结复方对IgA肾病肾内小动脉病变的防治及机制

IgA nephropathy is the glomerular disease with the highest incidence rate and the leading cause of end-stage renal failure. Therefore, to clarify the pathogenesis and find out the prevention and treatment of chronic progress are the current research focus. The incidence of renal artery disease in this case is extremely high, and it is related to the prognosis, but the pathogenesis remains to be studied. ShenfukangII is a compound of kidney reinforcing, dissolving, which would have a positive effect Clinical treatment to the disease. This study is to explore the possible mechanism rate of renal artery disease in IgA nephropathy, take the aldosterone and ERK/NF-κB as targets, and will launch: 1. Clinical research. Choose IgA nephropathy with/without vascular disease and non-IgA nephropathy patients as samples, determine the serum aldosterone, and observe the characteristics of renal minor arterial lesions in renal biopsy specimens. And then detect the expression of renal tissue, renal minor arteries, VEGF, MMP-9, PCNA, ERK1/2 and NF-κB. 2. In vivo experiment. Prepare IgA nephropathy rat models, and then detect the fore-mentioned index after 12 weeks while considering the intervention of losartan, eplerenone, sunflower capsules and ShenfukangII. 3. In vitro experiments. Use the serum of IgA nephropathy rat models and the fore mentioned containing serum to induce the nonstriated musle cells and endotheliocyte culture, and then detect ERK1/2 and phosphorylate of NF-κB. Try to reveal the function of ShenfukangII to the IgA nephropathy from the cell molecular level.

IgA肾病是发病率最高的肾小球疾病，是导致终末期肾衰竭的主要病种，阐明发病机理以防治慢性化进展是目前研究热点。本病肾内小动脉病变发生率极高且与预后相关而机理尚待研究。肾复康Ⅱ号是益肾散结复方，临床治疗本病具有肯定疗效。本课题拟探讨IgA肾病肾内小动脉病变的可能发生机制，以醛固酮及ERK/NF-κB为靶点，开展①临床研究：选取本病伴或不伴血管病变及非本病患者，测定血清醛固酮，观察肾活检标本中肾内小动脉病变特点并检测肾组织、肾内小动脉VEGF、MMP-9、PCNA、 ERK1/2、NF-κB的表达；②体内实验：制备IgA肾病大鼠模型，用氯沙坦、依普利酮、黄葵胶囊、肾复康Ⅱ号进行干预，12周后检测上述指标；③体外实验：通过用IgA肾病模型大鼠血清与上述分组含药血清共同干预醛固酮诱导的平滑肌细胞及内皮细胞培养，检测ERK 1/2、NF-κB磷酸化。从细胞分子水平揭示肾复康Ⅱ号对本病的作用机理。

IgA肾病（IgAN）是最常见原发性肾小球肾炎，是导致终末期肾病的重要原因。既往研究发现本病有不同程度肾内小动脉病变，与其预后密切相关。本研究引入本病成因与“肾气不足，痰瘀互结”相关，观察益肾散结方制剂防治IgAN及肾内小动病变作用机制。①临床研究：选取IgAN伴或不伴血管病变及非本病患者，测量肾穿刺标本中肾内小动脉内膜/血管外径值、中膜/血管外径值、管壁/血管外径值，检测肾组织VEGF、MMP-9、PCNA、ERK1/2、NF-κB表达，及尿蛋白定量、血肌酐、尿素氮、醛固酮、血管紧张素Ⅱ，结果显示IgAN伴血管病变患者血肌酐、尿素氮显著升高，ERK1/2、MMP-9、NF-κB、PCNA、VEGF表达较强，且各细胞因子与IgAN肾内小动脉内膜增厚、中膜增厚呈正相关。②实验研究：建立IgAN大鼠模型，用肾复康Ⅱ号、黄葵胶囊、氯沙坦钾、螺内酯、依普利酮干预，监测尿蛋白定量、血肌酐、尿素氮、醛固酮、血管紧张素Ⅱ，测大鼠肾组织小动脉厚度，及VEGF、MMP-9、PCNA、ERK1/2、NF-κB表达，结果显示肾复康Ⅱ号能显著降低大鼠尿蛋白定量、血肌酐、尿素氮、醛固酮，下调VEGF、MMP-9、PCNA、ERK1/2、NF-κB表达，与模型组相比肾复康Ⅱ组内膜/血管外径、管壁/血管外径有差异。IgAN肾内小动脉管壁增厚与VEGF、PCNA、MMP-9呈正相关，醛固酮与VEGF、PCNA、MMP-9表达均呈中度正相关。③细胞实验研究：通过不同药物含药血清干预醛固酮诱导的平滑肌细胞、内皮细胞，观察细胞增殖、生长周期情况，检测VEGF、MMP-9及PCNA表达及ERK（1/2）/NF-κB磷酸化情况，发现醛固酮促使平滑肌细胞异常增殖、内皮细胞凋亡，促进两种细胞ERK1/2、NF-κB磷酸化，上调平滑肌细胞VEGF、MMP-9及PCNA表达，下调内皮细胞VEGF、MMP-9及PCNA表达。阐明IgAN肾内小动脉病变机制可能与醛固酮分泌增多，启动ERK(1/2)/NF-κB信号通路，调节VEGF、MMP-9及PCNA表达相关，促进平滑肌细胞增殖，加快内皮细胞凋亡。本研究揭示肾复康Ⅱ号可能通过阻断ERK（1/2）/NF-κB信号通路，调节VEGF、MMP-9及PCNA表达，抑制平滑肌细胞异常增生，恢复内皮细胞增殖活力、抑制其凋亡，改善肾内小动脉病变，从而延缓本病进展。