加味防己黄芪汤对UUO大鼠纤维化干预作用及对JAK2/STAT3信号通路影响

Intervention effect of Jiawei Fangji Huangqi Decoction on fibrosis and JAK2/STAT3 signal pathway in UUO rats

关键词:  加味防己黄芪汤  肾间质纤维化  JAK2/STAT3  UUO

Key words:  Jiawei Fangji Huangqi Decoction  RIF  JAK2/STAT3  UUO

摘要:

[目的] 证实加味防己黄芪汤对于单侧输尿管梗阻（UUO）大鼠纤维化的干预作用以及对JAK2/STAT3信号通路的影响。[方法] 选取SPF级雄性SD大鼠共60只，构建UUO模型，随机分为假手术组、模型组、中药低剂量组、中药中剂量组、中药高剂量组、缬沙坦组，每组10只，术后第14天取材。观测肾脏脏器系数、血肌酐、尿素氮、24 h尿蛋白定量；苏木精-伊红染色法（HE）、马松（Masson）病理染色；Real-time qPCR法检测NF-κB、IκBa前后表达变化；Western Blot法检测JAK2、STAT3、p-JAK2、p-STAT3、E-钙黏蛋白（E-cadherin）、α-平滑肌肌动特异性蛋白（α-SMA）前后表达变化。[结果] 各组大鼠肾脏脏器系数、血肌酐、尿素氮、24 h尿蛋白定量、HE、Masson病理染色结果显示，药物干预组较模型组明显改善，且中剂量、高剂量组整体改善较为明显。Real-time qPCR结果显示，NF-κB、IκBa药物干预组同模型组相比表达水平明显下降（P<0.01）。Western Blot结果，E-cad、α-SMA、p-Jak2、p-Stat3蛋白表达情况，药物干预组同模型组相比具有明显改善（P<0.05），且中药中剂量、高剂量组较模型组具有明显的下调（P<0.05）。[结论] 加味防己黄芪汤能够改善UUO大鼠的血肌酐、尿素氮、24 h尿蛋白、减轻UUO大鼠的病理损伤、下调UUO大鼠E-cad、α-SMA、p-Jak2、p-Stat3、NF-κB、IκBa的蛋白表达，从一定程度上能够证实加味防己黄芪汤能够干预JAK2/STAT3信号通路起到抗炎、抗纤维化的作用，对于RIF具有延缓作用。

Abstract:

[Objective] To confirm the intervention effect of modified Fangji Huangqi Decoction on fibrosis in UUO rats and the effect on JAK2/STAT3 signaling pathway.[Methods] A total of 60 SPF male SD rats were selected to establish a UUO model and randomly divided into sham operation group, model group, low-dose Chinese medicine group, medium-dose Chinese medicine group, high-dose Chinese medicine group and valsartan group, with 10 rats in each group. The samples were collected 14 d after surgery. Renal weight/body weight coefficient, serum creatinine, urea nitrogen and 24 h urinary protein were observed. HE and Masson pathological staining;the expressions of NF-κB and IκBa were investigated by Real-time qPCR. The expressions of JAK2, STAT3, p-JAK2, p-STAT3, E-cadherin and α-SMA were detected by Western Blot.[Results] The results of renal/body weight coefficient, blood creatinine, urea nitrogen, 24 h urinary protein quantification, HE and Masson pathological staining of rats in each group showed that the drug intervention group was significantly improved compared with the model group, and the overall improvement was more obvious in the medium-dose and high-dose groups. Real-time qPCR results showed that the expression levels of NF-κB and IκBa in drug intervention group were significantly decreased compared with model group(P<0.01). Western Blot results showed that the protein expressions of E-cad, α-SMA, p-Jak2 and P-Stat3 in the intervention group were significantly improved compared with the model group(P<0.05), and the medium-dose and high-dose groups were significantly down-regulated compared with the model group(P<0.05).[Conclusion] Fangji Huangqi Decoction can improve the serum creatinine, urea nitrogen and 24 h urine protein of UUO rats, reduce the pathological injury of UUO rats, and down-regulate the protein expression of E-cad, α-SMA, p-Jak2, p-Stat3, NF-κB and I-κBa of UUO rats. To a certain extent, it can be confirmed that modified Fangji Huangqi Decoction can interfere with JAK2/STAT3 signaling pathway and play an anti-inflammatory and anti-fibrosis role, which has a protective effect on RIF.