Huoxue Jiedu Huayu recipe(活血解毒化瘀方)alleviates contralateral renal fibrosis in unilateral ureteral obstruction rats by inhibiting the transformation of macrophages to myofibroblast

OBJECTIVE:To investigate the action and underlying mechanisms of Huoxue Jiedu Huayu recipe(活血解毒化瘀方,HJHR)against unilateral ureteral obstruction(UUO)-induced injury in the contralateral kidney.METHODS:Forty-eight male Sprague-Dawley rats weighing(200±10)g were used in this study and randomly assigned to 4 groups:a sham group,a UUO group,a UUO+eplerenone(EPL)group,and a UUO+HJHR group.The contralateral kidneys were harvested for further study 180 d after surgery.Histological analysis,immunohistochemistry and immunofluorescence were used to study the fibrosis of the contralateral kidneys obtained from UUO rats.Contralateral kidney damagerelated pathway proteins were detected by real-time polymerase chain reaction and Western blot analysis.RESULTS:HJHR significantly inhibited fibrosis of the contralateral kidney in UUO rats by attenuating the UUOinduced macrophage-to-myofibroblast transition(MMT)in the contralateral kidney.Moreover,HJHR attenuated fibrosis in the contralateral kidney of UUO rats by preventing MMT through the aldosterone/mineralocorticoid receptor/serum/glucocorticoid regulated kinase 1 pathway.CONCLUSIONS:Our findings suggest that HJHR may be a potential treatment for renal interstitial fibrosis of obstructive nephropathy.

益气活血解毒中药及肾上腺髓质素抑制醛固酮诱导的HK2细胞自噬作用研究

目的:观察益气活血解毒中药血清及肾上腺髓质素(Adrenomedullin,ADM)抑制醛固酮(Aldosterone,ALD)诱导的人近端肾小管上皮细胞HK2自噬的作用。方法:本实验采用1μmol/L的ALD诱导HK2细胞自噬,给予10%的益气活血解毒含药血清及100 nmol/L的ADM治疗。实验分为对照组(CON组),ALD组,ALD+益气活血解毒含药血清组(ALD+TCM组),ALD+ADM组,48 h后收集细胞并观察变化。采用免疫细胞荧光、Western blot法检测血清糖皮质激素诱导蛋白激酶1 (Serum and glucocorticoid induced kinase,SGK-1)、磷酸化细胞外信号调节激酶1/2 (Phosphorylated extracellular signal-regulated protein kinase,P-ERK1/2)、磷酸化哺乳动物雷帕霉素靶蛋白(Phosphorylated mammalian target of rapamycin,P-mTOR)/mTOR、自噬相关基因5(Autophagy associated gene,Atg5)、酵母自噬基因6 (Atg6)的同源物Beclin1、自噬标志微管相关蛋白1轻链3 (Microtubularassociated protein 1 light chain 3,LC3)的表达。结果:与CON组比较,ALD组SGK-1、P-ERK1/2、Atg5、LC3和Beclin1表达明显增强(P <0.01),P-mTOR/mTOR表达减弱(P <0.01);与ALD组比较,ALD+TCM组、ALD+ADM组这些指标表达均减弱(P <0.01),P-mTOR/mTOR表达增强(P <0.01)。结论:益气活血解毒中药、ADM均可抑制ALD诱导的HK2细胞自噬,从而减轻肾小管的损伤,减缓肾脏损伤的进程。