化瘀通络中药对糖尿病肾病大鼠肾脏足细胞骨架蛋白的干预作用

Effect of Huayu Tongluo Traditional Chinese Medicine on Skeleton Protein in Podocytes of Rats with Diabetic Nephropathy

通过化瘀通络中药对糖尿病肾病(DN)大鼠肾脏足细胞骨架蛋白α-actinin-4、Synaptopodin的干预作用,探讨化瘀通络中药减少蛋白尿,防治DN的可能作用机理。选取健康雄性SD大鼠50只并按随机数字表法分为正常组(C组)10只,其余40只采用高糖高脂饲料联合1%链脲佐菌素(STZ)腹腔注射复制糖尿病(DM)大鼠模型,72 h后尾静脉取血检测血糖,血糖≥16.7 mmol/L为糖尿病大鼠造模成功,成模大鼠再随机分为3组,分别为模型组(M组)、化瘀通络中药组(Z组)、厄贝沙坦组(I组)。各治疗组分别给予不同干预,16周后分别检测各组大鼠24 h尿蛋白(24 h UTP)、血凝及血小板参数等指标;采用蛋白质印迹法(Western-Blot)、实时荧光定量(Real-time PCR)方法检测各组大鼠肾脏足细胞骨架蛋白α-actinin-4、Synaptopodin的表达情况。结果显示,与C组比较,其余各组大鼠24 h UTP明显升高(P<0.01),血凝、血小板相关指标均出现明显变化;与M组相比,24h UTP均明显减少(P<0.01),血凝、血小板相关指标均有明显改善;与I组比较,中药组大鼠24 h UTP明显减少(P<0.05)。与C组相比,其他各组大鼠肾组织α-actinin-4、Synaptopodin表达水平均明显降低(P<0.05,P<0.01);与M组相比,两治疗组两种蛋白表达均明显升高(P<0.05,P<0.01);与I组比较,中药组大鼠Synaptopodin的基因和蛋白表达均明显上调(P<0.05)。以上结果说明化瘀通络中药可以降低糖尿病肾病大鼠尿蛋白,其机制可能与改善血液高粘、高凝状态,上调足细胞骨架蛋白α-actinin-4、Synaptopodin,维持足细胞正常结构和功能有关。

To explore the intervention effect of Huayu Tongluo traditional Chinese medicine on diabetic nephropathy （DN） and its effect on skeleton protein. Ten rats were randomly selected as the normal group （ C group ） from 50 male Sprague-Dauley rats, the other 40 rats were injected with 1% streptozotocin （35 mg/kg） after fed high-fat and high-glucose diet for one month. The successful model rats （ blood glucoses〉 16.7 mmoL/L） were randomly divided into three groups：model group （M group ）, Huayu Tongluo traditional Chinese medicine group （Z group ）, Irbesartan group （I group）. Each treatment group was given corresponding intervention, once a day for 16 weeks. At the end of the 16th week ,24 h UTP,blood coagulation and platelet parameters were tested. The protein and mRNA expressions of α-actinin- 4 and Synaptopodin in kidney tissue were examined by Western-blot and Real-time PCR methods. The results showed that 24 h UTP was significantly increased in other groups （P 〈 0.01 ） compared with C group, and in two treatment groups it was also significantly improved （ P 〈 0. 01 ） compared with M group, and it reduced significantly in Z group than that in Ⅰ group （P 〈 0. 05）. Similar results appeared that compared with C group, the expression of α-actinin-4 and Synaptopodin reduced in other groups （P〈0.01）,it was significantly higher in two treatment groups （P 〈0.01 ） than that in M group, and expression in Z group was significantly higher （ P 〈 0.05 ） than that in Ⅰ group. The results indicated that Huayu Tongluo traditional Chinese medicine can reduce urinary protein in DN rats. The mechanism may be related to improve the high blood viscosity, hypercoagulable state and up regulating the expression of a-actinin-4 and synaptopodin. Key words ： diabetic nephropathy; Huayu-tongluo traditional Chinese medicine; urine protein; skeleton protein