抗纤灵方对单侧输尿管梗阻大鼠肾组织肝细胞生长因子mRNA及细胞外信号调控蛋白激酶1/2和p38磷酸化的影响

Effect of Kangxianling Decoction on expression of hepatocyte growth factor mRNA and phosphorylations of extracellular signal-regulated protein kinase 1/2 and p38 in renal tissue of rats with unilateral ureteral obstruction

目的:研究中药复方抗纤灵方对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)所致肾纤维化大鼠肝细胞生长因子(hepatocyte growth factor,HGF)基因表达和丝裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)通路的影响,初步探讨其发挥疗效的作用机制。方法:雄性SD大鼠18只随机分为3组,每组6只。用单侧输尿管结扎术建立大鼠肾纤维化模型,抗纤灵灌胃给药2周后检测血肌酐(serum creatinine,Scr)、血尿素氯(blood urea nitrogen,BUN)及梗阻肾羟脯氨酸含量;采用逆转录-多聚酶链反应(reverse transcription polymerase chain reaction,RT-PCR)检测HGF mRNA表达;采用蛋白免疫印迹法检测其受体c-Met蛋白表达及MAPK通路信号分子细胞外信号调控蛋白激酶1/2(extracellular signal-regulated protein kinase 1/2,ERK1/2)和p38磷酸化情况。结果:与假手术组比较.模型组大鼠Scr、BUN及梗阻肾羟脯氧酸含量均显著增高;肾组织HGF mRNA水平显著下调,c-Met蛋白表达显著上调,且ERK1/2和p38明显磷酸化。中药干预后,BUN及羟脯氨酸含量显著降低,肾组织HGF mRNA水平明显上调,ERK1/2和p38磷酸化被显著抑制。结论:抗纤灵方可通过增加肾纤维化保护性因子HGF mRNA的表达及抑制肾组织MAPK通路分子ERK1/2和p38磷酸化以改善肾间质纤维化大鼠肾功能,减少其胶原含量。

Objective： To study the effect of Kangxianling Decoction （KXLD）, a compound traditional Chinese herbal medicine, on expression of hepatocyte growth factor （HGF） mRNA and phosphorylations of extracellular signal-regulated protein kinase 1/2 （ERK 1/2） and p38 in renal tissue of rats with unilateral ureteral obstruction （UUO）. Methods： Eighteen male SD rats were randomly divided into 3 groups： sham-operated group, untreated group and KXLD-treated group. A rat model of renal interstitial fibrosis was established by UUO. Rats with UUO were sacrificed after intragastric administration of KXLD for 14 days, and the parameters such as serum creatinine （SCr）, blood urea nitrogen （BUN） .and hydroxyproline in the kidney of rats in 3 groups were analyzed. The expression of HGF mRNA in kidney tissue was determined by reverse transcription polymerase chain reaction. The expressions of c-Met protein, ERK1/2 protein, p38 protein and the phosphorylations of ERK1/2 and p38 were determined by Western blotting method. Results： The levels of SCr, BUN and hydroxyproline in the untreated group were significantly increased as compared with those in the sham-operated group （P〈0.05）. The expression of HGF mRNA in the untreated group was significantly down-regulated. The expression of c-Met protein and the phosphorylations of ERK1/2 and p38 in the kidney tissue of rats with UUO in the untreated group were significantly up-regulated. After intervention with KXLD, the phosphorylations of ERK1/2 and p38 were all significantly inhibited except for c-Met expression. The HGF mRNA was increased in KXLD-treated group. Conclusion： KXLD can decrease the level of collagen in the obstructed kidney of rats with UUO and alleviate the renal interstitial fibrosis in rats with UUO through enhancing the HGF mRNA expression and inhibiting the phosphorylations of ERK1/2 and p38.