早期糖尿病肾病“气阴两虚,络脉瘀阻”病机与AGE-RAGE轴相关性研究

The Study of Relationship between Material Bases of Early Diabetic Nephropathy as Deficiency of both Qi and Yin and Obstruction of Collat-erals and AGE-RAGE

[目的]探讨糖基化终末产物(advanced glycation end products,AGE)及其受体(receptor of AGE,RAGE)与早期糖尿病肾病(diabetic nephropathy,DN)大鼠"气阴两虚、络脉瘀阻"病机的关系。[方法]高脂高糖饲料联合1%链脲佐菌素(streptozotocin,STZ)30 mg·kg-1静脉注射复制早期DN大鼠模型,将造模成功的大鼠随机分为DN模型组、糖肾方低剂量组、糖肾方中剂量组、糖肾方高剂量组、缬沙坦组,同时设正常组。观察大鼠一般情况,检测空腹血糖(fasting blood glucose,FBG)、肌酐清除率(creatinine clearance,Ccr)等指标;ELISA技术检测尿白蛋白(urine albumin,U-alb)的浓度及血清中AGE的含量;分别用Real-time PCR、Western blot法检测肾组织中RAGE的m RNA及蛋白表达;血液流变仪检测血浆粘度和全血还原黏度;PAS染色观察肾脏病理。[结果]DN模型组大鼠精神萎靡、懒动、体型消瘦,出现多饮、多食、多尿症状,粪质稀软,毛色枯黄,竖立。DN模型组FBG升高,血浆粘度、全血还原黏度增加,U-alb排泄率增加,Ccr降低,AGE、RAGE明显升高。经糖肾方及缬沙坦干预后一般情况改善;U-alb排泄率有不同程度的降低,其中糖肾方中、高剂量组、缬沙坦组明显降低;Ccr有一定程度的升高,其中糖肾方高剂量组和缬沙坦组明显升高;血浆粘度和全血还原黏度降低;AGE含量降低,RAGE表达明显下降;血清中AGE含量与肾组织中RAGE蛋白表达、U-alb排泄率间呈正相关(r值分别为0.637、0.622),与Ccr呈负相关(r值为-0.456)。[结论]早期DN大鼠存在"气阴两虚、络脉瘀阻"病机,AGE-RAGE与早期DN"气阴两虚、络脉瘀阻"的病机可能具有相关性。

[Objective] To investigate the relationship between the AGE-RAGE and Qi and Yin deficiency and blood stasis of pathogenesis in diabetic nephropathy（DN）.[Methods] The model was established by intravenous injection low-dose streptozotocin（STZ, 30 mg·kg-1） after having the high fat/high glucose diets for one month. And then that were divided into DN group, Tangshenfang low, medium, high groups, Valsartan group. Fasting blood-glucose （FBG） and Creatinine clearance（Ccr） were monitored. The morphological changes of renal tissue were examined under microscopy on sections stained with PAS. The RAGE mRNA and protein expression were determined by Real-time PCR and Western blot respectively; the urine album（U-alb） and content of AGE in serum were determined by ELISA. [Results] The model group rats acted obviously as deficiency of both Qi and Yin and obstruction of collaterals. compared with normal group, FBG, urinary albumin excretion rate and the content of AGE, RAGE expression were markedly increased; Ccr was markedly decreased in DN model group; blood viscosity and whole blood reduction viscosity were increased. FBG, Urinary albumin excretion rate and the content of AGE, RAGE expression were markedly decreased, Cer was markedly increased by the intervention of Tangshenfang and valsartan. Blood viscosity and whole blood reduction viscosity were decreased; it was obviously and positively correlated between AGE with RAGE expression and U-alb, excretion rate and negatively correlated with the Ccr, r value was -0.456. [Conclusion] The early DN model rats were in the pathogenesis of Qi and Yin deficiency and blood stasis. And the AGE-RAGE may have close relationship with the pathogenesis of Qi and Yin deficiency and blood stasis.