Intervention effects of Compound Houttuyniae Herba to diabetic renal damage based on SOCS-JAK/STAT negative feedback regulation
Article
Figures
Metrics
Preview PDF
Reference
Related
Cited by
Materials
摘要:
目的：糖尿病肾病（diabetic nephropathy, DN）是糖尿病（Diabetic mellitus，DM）临床常见的慢性并发症之一，本实验旨在研究复方鱼腥草三种提取部位及其完整成分对db/db小鼠肾损害的保护作用，并通过探究各提取物对JAK2/STAT3通路及其下游蛋白的影响来探讨复方鱼腥草防治糖尿病肾病的可能机制.方法：1、提取复方鱼腥草水提物、醇提物、挥发油成分,并将三种成分按照所得质量比配制得到复方鱼腥草完整成分。2、选取8只db/m小鼠为正常组；56只db/db小鼠分为模型组、盐酸二甲双胍组、AG490组、复方鱼腥草水提物组、复方鱼腥草醇提物组、复方鱼腥草挥发油组以及复方鱼腥草完整成分组，每组8只，给药8周，记录小鼠体重以及血糖，8周后检测小鼠血糖、肾功能指标及肾脏指数，酶联免疫试剂盒法检测血浆胰岛素水平（FINS）、转化生长因子-β1（TGF-β1）、纤连蛋白（FN）、HE染色观察小鼠肾脏病理变化。3、RT-PCR法检测各组小鼠肾组织中的JAK2mRNA、STAT3mRNA，SOCS-1mRNA表达的变化情况； Western blot法检测肾组织中的JAK2、p-JAK2、STAT3、p-STAT3、SOCS-1、c-fos及c-jun蛋白表达变化，运用免疫荧光法检测肾组织c-fos、c-jun蛋白的表达。结果：1、给药八周后，与正常组比较，模型组db/db小鼠血浆转化生长因子-β1（TGF-β1）、纤连蛋白（FN）、治疗组小鼠血清胰岛素水平、胰岛素敏感指数较模型组升高，转化生长因子-β1（TGF-β1），纤连蛋白（FN），水平降低（P<0.05），各治疗组小鼠肾脏组织病理改善明显。2、模型组小鼠肾组织p-JAK2，p-STAT3及JAK2 mRNA，STAT3mRNA的表达均对比正常组小鼠肾组织的表达明显增高(P<0.05)；与模型组相比，各给药组小鼠肾组织p-JAK2，p-STAT3表达均有降低，挥发油组及完整成分组小鼠肾组织SOCS-1基因及蛋白明显升高（p<0.05）；各给药组的JAK2、STAT3基因表达及蛋白变化不显著。各治疗组的c-fos、c-jun在肾小管及肾间质的表达均有不同程度下降。结论：复方鱼腥草对糖尿病肾损伤具有改善作用，其保护肾脏的作用可能与降低尿蛋白排泄，调节血糖，维持肾脏结构、功能的完整性有关。其改善糖尿病肾损伤的分子机制可能通过SOCS 负反馈调节JAK/STAT信号转导通路相关基因及蛋白表达，从而抑制降低下游原癌基因c-fos、c-jun的表达，减少炎症因子的活化，降低FN的分泌，减少ECM聚积有关。
Abstract:
Objective To research the protective effects of different extracts from Compound Houttuyniae Herba (CHH) and its mechanism through JAK/STAT-SOCS-1 signaling pathway. Methods The normal group comprised db/m mice (n = 8). db/db mice were randomly divided into seven groups with eight mice in each group according to the applied treatment method: model group, metformin hydrochloride (MH) group, AG490 group, water extract (WE) group, ethanol extract (EE) group, volatile oil (VO) group, and mixture (MG) group (mixture of above three extract) of CHH. After 8 weeks, the general status, biochemical indicators, and renal histological changes in the mice were evaluated, the total RNA and protein were collected and RT-PCR method was used to examine the effect on mRNA expression of JAK2, STAT3, SOCS1, and Western blot method was used to detect the protein expression of JAK2, P-JAK2, STAT3, P-STAT3, SOCS1, c-fos, and c-jun. Immunofluorescence was used to observe the protein expression of c-fos and c-jun in kidney tissue. Results Compared with normal group, the serum level of TGF-β1, FN of EE, VO, and MG groups were decreased (P < 0.05). Renal function was also improved but not significantly. Also, the renal histology was improved especially in the mixture group. The protein expression of JAK2, STAT3, P-JAK2, P-STAT3, and the genetic expression of JAK2 and STAT3 in kidney tissue were significantly increased in the model group (P < 0.05). Compared with the model group, the expression of P-JAK2 and P-STAT3 was down-regulated in treatment groups; The expression of SOCS-1 of VO and mixture groups were elevated (P < 0.05). Conclusion CHH has beneficial effects on diabetic renal injury, which may protect and improve the kidney function and reduce urinary protein, maintain the integrity of the kidney structure and function.
关键词:
复方鱼腥草;糖尿病肾病;JAK/STAT-SOCS;c-fos;c-jun
Keywords:
Project Supported:
National Natural Science Foundation of China (No. 81403345) and Innovation Program of Shanghai Municipal Education Commission (No. 14YZ059)
