妊娠激活醛固酮刺激肾小管上皮细胞增殖加重慢性肾脏病损伤机制以及益肾健脾泻浊解毒化瘀中药的治疗作用

Pregnancy is an independent risk factor of chronic kidney disease which activated aldosterone plays important role in it. Pregnancy could activate aldosterone, not only induce changes in gestational hypertension, but also stimulate kidney cells proliferation and aggravate kidney damage. Results from previous studies which Huayuditantongluo herbs inhibit cell proliferation have confirmed that unilateral ureteral ligation (UUO) was used to induce activated aldosterone and inflammation in renal interstitial fibrosis. In this study, “deficiency of kidney and spleen and pathogenic damp accumulation” as traditional Chinese pathogenesis, activated aldosterone-SGK-1/ERK-cell proliferation-ECM accumulation as research approaches, we will research the mechanism of kidney injury aggravated by pregnancy. UUO is used to induce kidney injury. After 8 weeks, the rats are caged together and observed dynamic changes of pregnancy blood pressure, renal function and aldosterone. PCNA is detected and the role of SGK-1/NF-kB/P-ERK signaling pathway in promoting cell proliferation is discussed, at the same time aldosterone receptor blockers and herbs of YishenJianpi xiezhuohuayujiedu are given. The effect of Chinese herbs will be observed, whether can inhibite aldosterone secretion, down-regulate SGK-1/ERK, future inhibite cell proliferation, and provide experimental basis for renal injury caused by pregnancy.

妊娠是加重慢性肾脏病损伤的独立危险因素，醛固酮活化在其中发挥关键作用。妊娠可激活醛固酮，作用于肾脏，不仅诱导血压改变，亦可刺激肾脏固有细胞增殖，加重肾脏损伤。在前期研究化瘀涤痰通络中药抑制细胞增殖的过程中，发现单侧输尿管梗阻可激活醛固酮，介导炎性损伤，促进纤维化进展。在此基础上，本研究以“肾脾亏虚，浊毒内蕴”为病机，以醛固酮活化-SGK-1/ERK-细胞增殖-ECM积聚为途径，探讨妊娠加重肾损伤的可能机制。结扎雌性未经产Wistar大鼠输尿管，8周后与雄鼠合笼制备肾脏损伤实验动物妊娠模型，动态观察血压、肾功能及醛固酮改变，检测细胞增殖标志物PCNA的表达，解析SGK-1/NF-κB/P-ERK信号通路促进细胞增殖的作用。同时给以醛固酮受体阻断剂及益肾健脾泻浊解毒化瘀中药，观察其抑制醛固酮活化、下调SGK-1/ERK表达、抑制细胞增殖、保护肾功能的效应，为临床预防妊娠加重肾脏损伤提供实验。

妊娠是加重慢性肾脏病损伤的独立危险因素，醛固酮活化在其中发挥关键作用，醛固酮被激活后可介导肾脏炎性损伤，促进纤维化进展。在此基础上，本研究以“肾脾亏虚，浊毒内蕴”为病机，以醛固酮活化-SGK-1/P-ERK-细胞增殖-ECM积聚为途径，探讨妊娠加重肾损伤的可能机制。雌性未经产Wistar大鼠随机取 60只采用UUO手术造成肾脏损伤，其余 30 只大鼠作为对照组，仅将输尿管游离但不结扎离断。对照组和 UUO 造模大鼠再各自分为2 组，1 组与雄性大鼠合笼，另 1 组不加处理。将 UUO 造模大鼠随机分为 UUO 组（UUO group）、模型组（UP group）、西药组（EPL group）、中药组（TCM group）；对照组大鼠分为假手术组（Sham group）、假手术＋妊娠组（SP group），共 6 组，每组 15 只。UUO术后给药，EPL组给以依普利酮 100mg.kg-1 .d -1 从饲料中加入，TCM组给以益肾健脾泻浊解毒化瘀中药煎剂灌胃 13.7g.kg-1 .d -1，其余 4 组给予等容量蒸馏水灌胃，连续给药至妊娠第 19 天，处死母鼠，留取血清，摘取梗阻侧肾脏，以备指标检测。结果显示：UP组多数大鼠动情周期紊乱，见栓怀孕率低，流产情况多。血清和组织醛固酮含量明显升高（P＜0.01），血清P、E2表达水平明显降低（P＜0.01）。肾脏病理学检测可见大量炎性细胞浸润，肾小管明显扩张，间质胶原成分显著增多；肾功能检测BUN、Scr明显升高，Ccr明显降低（P＜0.01）；PCNA、SGK-1、P-ERK、NLRP3、MCP-1、Caspase-1、IL-1β、NF-κB的蛋白表达明显增强，主要表达于肾小管以及肾间质。依普利酮及中药可减轻改善肾功能，肾脏病理损伤；降低血清醛固酮含量（P＜0.01），提高P、E2含量（P＜0.01, P＜0.05）；下调PCNA、SGK-1、P-ERK以及NLRP3、MCP-1、Caspase-1、IL-1β、NF-κB蛋白水平的表达（P＜0.01, P＜0.05）。结果表明益肾健脾泻浊解毒化瘀中药可通过阻断醛固酮活化，下调 SGK-1/P-ERK表达，抑制肾小管上皮细胞过度增殖，还可下调NLRP3 /caspase-1/IL-1β表达，减轻炎性坏死，减少 ECM 集聚，从而起到拮抗RIF作用。