基于miRNA靶向SPRED-1/PIK3R2调控VEGF-Notch信号通路研究肾纤维化"脉络瘀阻"及“益肾活血法”干预机制

Renal fibrosis is the final common pathway of the chronic kidney disease developing to end stage kidney disease and the hypoxic/ischemic injury caused by pathological changes of renal interstitial capillary plays the key role. Promoting the restoration of the injured renal interstitial capillary network is the key to antagonize renal interstitial fibrosis.The UUO rats were used as experimental model in this study and the differential expression of renal angiogenesis related miRNAs were screened after they were intragastrically administrated with Yishenhuoxue prescription and losartan; at the same time, the hypoxia induced culturing of RMVEC was performed in vitro. After the intervention of pastille serum and losartan, miRNA126, SPRED1/PIK3R2,VEGF,VEGFR-2,Notch1 mRNA and protein expression of the kidney and RMVEC were respectively detected;MVD was detected with immunohistochemistry and the ability of RMVEC canalization was also observed in this study.The RMVEC were transfected in vitro and the effects of inhibited miRNA126 on angiogenesis were observed. This study was to explore the biology nature of renal fibrosis "stasis resistance in vein blood" from molecular level and clarify the mechanism of "kidney benefiting and blood circulation activation" by intervening miroRNA and regulating VEGF-Notch signal path to promote microvascular, which could provide new ideas and methods for microcosmic syndrome differentiation and traditional Chinese medicine treatment for chronic renal fibrosis.

肾纤维化是所有CKD发展至终末期肾脏病的最后共同通路，肾间质微血管病变所导致的低氧/缺血性损伤在慢性肾脏病中起关键作用，促进已损伤肾间质毛细血管网的修复，是拮抗肾脏间质纤维化的关键。本研究以UUO大鼠为实验模型，予益肾活血方、氯沙坦灌喂给药，筛选肾血管新生相关miRNAs差异表达，同时体外低氧诱导培养RMVEC，予含药血清、氯沙坦进行干预，分别检测肾脏及RMVEC 的miRNA126、SPRED1/PIK3R2、VEGF、VEGFR-2、Notch1 mRNA及蛋白表达；免疫组化学检测MVD；观察RMVEC细胞成管能力；体外进行RMVEC转染，观察miRNA126抑制后对血管新生的影响。本研究旨在从分子水平探索肾纤维化"脉络瘀阻"的生物学本质；阐明"益肾活血方"通过干预miroRNA调控VEGF-Notch信号通路发挥促微血管新生机制，为慢性肾纤维化的"微观辨证"及中医药治疗提供新的思路。

肾纤维化是所有CKD发展至终末期肾脏病的最后共同通路，肾间质微血管病变所导致的低氧/缺血性损伤在慢性肾脏病中起关键作用，促进已损伤肾间质毛细血管网的修复，是拮抗肾脏间质纤维化的关键。本研究以UUO大鼠为实验模型，予益肾活血方、氯沙坦灌喂给药，筛选肾血管新生相关miRNAs差异表达，同时体外低氧诱导培养RMVEC，予含药血清、氯沙坦进行干预，分别检测肾脏及RMVEC的miRNA126、SPRED1/PIK3R2、VEGF、VEGFR-2、Notch1mRNA及蛋白表达；免疫组化学检测MVD；观察RMVEC细胞成管能力；体外进行RMVEC转染，观察miRNA126抑制后对血管新生的影响。本研究旨在从分子水平探索肾纤维化"脉络瘀阻"的生物学本质；阐明"益肾活血方"通过干预miroRNA调控VEGF-Notch信号通路发挥促微血管新生机制，为慢性肾纤维化的"微观辨证"及中医药治疗提供新的思路