健脾补肾泻浊方对妊娠加重慢性肾病肾脏ERK/NF-κB信号通路及细胞增殖的影响

Effects of Jianpi Bushen Xiezho Formula on the Renal ERK/NF-κB Signaling Pathway and Cell Proliferation in Pregnancy-Aggravated Chronic Kidney Disease

目的:观察健脾补肾泻浊方抑制妊娠加重慢性肾脏病大鼠肾脏细胞增殖的作用及机制。方法:健康雌性未经产Wistar大鼠48只随机分为假手术未孕组、假手术妊娠组、模型未孕组、模型妊娠组、依普利酮0.1 g/kg组、健脾补肾泄浊方13.7 g/kg组,每组8只。除假手术未孕组和假手术妊娠组大鼠外,其余4组大鼠采用单侧输尿管结扎(Unilateral ureteral obstruction, UUO)手术复制肾间质纤维化模型,两个治疗组分别给予依普利酮和健脾补肾泻浊方治疗,8 w后把处于动情周期的雌鼠(假手术妊娠组、模型妊娠组、依普利酮0.1 g/kg组、健脾补肾泄浊中药13.7 g/kg组)与雄鼠进行合笼,妊娠之后第19 d处死母鼠,取血清标本,摘取梗阻对侧肾脏。检测大鼠的肾功能和导致肾间质纤维化的相关指标。结果:与假手术妊娠组比较,模型对照组大鼠肾脏有明显的胶原纤维沉积和大量炎性细胞浸润,肾小管扩张明显,血清肌酐(Serum creatinine, Scr)、尿素氮(Blood urea nitrogen, BUN)、醛固酮(Aldosterone, ALD)、肿瘤坏死因子-α(Tumor necrosis factor,TNF-α)、白介素-1(Interleukine-1,IL-1)的含量显著升高(P<0.01),大鼠肾脏组织中增殖细胞核抗原(Proliferating ell nuclear antigen, PCNA)、核因子κB(Nuclear factor-kappa B,NF-κB)、细胞外信号调节激酶(Extracellular Signal regulated Kinase, ERK)、磷酸化细胞外信号调节激酶(Phospho-extracellular signal-regulated kinase, P-ERK)的蛋白表达显著上调(P<0.01)。与模型妊娠组比较,依普利酮0.1 g/kg组和健脾补肾泻浊方13.7 g/kg组胶原纤维的沉积明显减轻,炎性细胞的浸润明显减轻,血清Scr、BUN、ALD、TNF-α、IL-1的含量显著降低(P<0.01),肾脏组织中PCNA、NF-κB、ERK、P-ERK的蛋白表达显著下调(P<0.01)。结论:健脾补肾泻浊方可通过调控ERK/NF-κB信号传导通路所导致的细胞增殖来减少细胞外基质(Excetral Cellular Matrix, ECM)的沉积,延缓妊娠加重慢性肾脏病损伤的进展。

Objective:To observe the inhibitory effects and mechanism of the Jianpi Bushen Xiezho Formula(JPBSXZ) on the proliferation of kidney cells in rats with chronic kidney disease aggravated by pregnancy. Methods:Forty-eight healthy unpregnancy Wistar rats were randomly divided into two sham operation groups(unpregnancy and pregnancy),two model groups(unpregnancy and pregnancy),an eplerenone(0.1 g/kg) group, and a JPBSXZ(13.7 g/kg) group, with eight rats in each group. Rats, except for those in the two sham operation groups, were treated with unilateral ureteral ligation(UUO) to induce the model of renal interstitial fibrosis. The groups with drug intervention were administered correspondingly. After eight weeks of treatment, the female rats in the estrus cycle(the sham operation-pregnancy group, the model-pregnancy group, the eplerenone group, and the JPBSXZ group) were caged with male rats. On the 19 th day after pregnancy, the pregnant rats were sacrificed. Serum samples were taken and the contralateral kidneys were removed. The renal function of the rats and the indexes related to renal interstitial fibrosis were detected. Results:Compared with the sham operation-pregnancy group, the model-pregnancy group showed obvious collagen fiber deposition, infiltration of a large number of inflammatory cells, renal tubule dilation, increased content of serum creatinine(Scr), blood urea nitrogen(BUN), aldosterone(ALD), tumor necrosis factor-α(TNF-α), and interleukin-1(IL-1)(P<0.01),and up-regulated protein expression of proliferating cell nuclear antigen(PCNA), nuclear factor-kappa B(NF-κB), extracellular signal-regulated kinase(ERK), phosphorylated extracellular signal-regulated kinase(P-ERK) in kidney tissues(P<0.01). Compared with the model-pregnancy group, the eplerenone group and the JPBSXZ group displayed reduced deposition of collagen fibers, relieved inflammatory cell infiltration, decreased serum levels of Scr, BUN, ALD, TNF-α, and IL-1(P<0.01), and down-regulated expression of PCNA,NF-κB,ERK,and P-ERK in kidney tissues(P<0.01). Conclusion: JPBSXZ can reduce the deposition of extracellular matrix(ECM) by regulating the cell proliferation caused by the ERK/NF-κB signaling pathway to delay the progression of pregnancy-aggravated chronic kidney disease.