当归补血汤对糖尿病肾病大鼠PERK通路的影响

Effect of Danggui Buxue decoction on the expression of GRP78-PERK-eIF2α in diabetic nephropathy rats

关键词:  当归补血汤  内质网应激  PERK通路  糖尿病肾病

Key words:  Danggui Buxue decoction  endoplasmic reticulum stress  PERK pathway  diabetic nephropathy

摘要:

[目的]通过观察当归补血汤干预糖尿病肾病（DN）大鼠的葡萄糖调节蛋白78（GRP78）、蛋白激酶R样内质网激酶（PERK）、真核生物翻译起始因子2α（eIF2α）表达的变化，探讨其对防治DN的作用机制。[方法]实验设对照组、模型组、格列齐特组、当归补血汤低剂量组（低剂量组）、当归补血汤高剂量组（高剂量组），每组10只大鼠。以腹腔注射链脲佐菌素（STZ）联合高脂饲料建立DN大鼠模型，连续喂养8周，光镜和电镜观察肾脏结构变化，酶偶联比色法测血脂、血糖、肾功能相关指标；免疫组化和Western Blot测肾组织中GRP78、PERK、eIF2α蛋白的表达，Real-time PCR测GRP78的mRNA表达。[结果]模型组大鼠一般情况差，肾功能明显受损，肾脏结构严重损伤，GRP78、PERK、eIF2α蛋白及mRNA的表达明显高于对照组；当归补血汤治疗后肾功能及肾结构改善，GRP78、PERK、eIF2α的蛋白及mRNA表达明显下降，高剂量组下降更明显，呈浓度依赖性。[结论]当归补血汤能抑制模型组大鼠肾脏GRP78、PERK、eIF2α的表达，缓解内质网应激，具有保护肾功能作用。

Abstract:

[Objective] To observe the effects of Danggui Buxue decoction on the expression of glucose-regulated protein 78(GRP78), protein kinase R-like ER kinase (PERK), eukaryotic translation initiation factor 2 alpha(eIF2α) in diabetic nephropathy rats and explore the mechanism of its effect on the prevention and treatment of DN.[Methods] Fifty rats were randomly divided into blank group(N), model group, gliclazide gloup, low and high dose of Dangguibuxue decoction (DL/DH). The rat model of DN was established by intraperitoneal injection of streptozotocin (STZ) combined with high fat diet. After 8 weeks, the changes of renal structure were observed under light microscope and electron microscope;the serum glucose,lipids and the renal function related parameters were detected by enzyme coupling colorimetric method; the expression of GRP78, PERK and eIF2α protein in renal tissue were detected by immunohistochemistry and western blot and the mRNA expression of GRP78, PERK and eIF2α was detected by Real-time PCR.[Results] The rats in the DN group were in poor condition, the renal function and the kidney structure were seriously damaged, the expression of GRP78, PERK and eIF2α protein and mRNA in the DN group were significantly worse than those in the N group, the expression of GRP78,PERK and eIF2α protein and mRNA decreased significantly after treatment, the DH group decreased more significantly in a concentration-dependent manner.[Conclusion] Danggui Buxue decoction can inhibit the expression of GRP78, PERK and eIF2α in DN rats kidney, meanwhile, it can relieve endoplasmic reticulum stress and protect renal function.