基于microRNAs与炎症因子相关性探讨丹蛭降糖胶囊对GK大鼠糖尿病大血管病变治疗作用机制

Mechanism of Danzhi Jiangtang Capsules in treatment of diabetic macrovascular disease in GK rats based on correlation between microRNAs and inflammatory factors

探讨丹蛭降糖胶囊对Goto-Kakizaki(GK)大鼠糖尿病大血管病变治疗作用机制。采用高脂高糖联合内皮型一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸甲酯(L-NAME)(0. 1 g·L-1·d-1)喂养复制糖尿病大血管病变大鼠模型。按随机排列表将模型大鼠随机分为模型组、丹蛭降糖胶囊组(1 260,630,320 mg·kg-1)、阿托伐他汀组(105 mg·kg-1)、二甲双胍组(10 mg·kg-1),每组12只,连续灌胃给药8周。取Wistar大鼠12只作为正常对照组。观测大鼠体质量,饮水量,血糖及血浆TC,TG,LDL-C,HDL-C,IL-1β,IL-6,TNF-α,NO,ET-1变化;取主动脉组织,HE染色法观察病理改变;RT-PCR法检测大鼠主动脉IL-1β,IL-6,TNF-αmRNA水平,茎环RT-PCR法检测大鼠血浆与主动脉miRNA-126,miRNA-155,miRNA-146a,miRNA-21水平,对miRNAs与炎症因子之间进行典型相关性分析。结果显示,丹蛭降糖胶囊能增加大鼠体质量,降低饮水量,降低随机血糖,逆转大鼠主动脉组织损伤,降低血浆TC,TG,LDL-C,LDL-C,ET-1,IL-1β,IL-6,TNF-α,miRNA-155,miRNA-146a,miRNA-21水平,升高血浆HDL-C,NO含量,降低主动脉IL-1β,IL-6,TNF-αmRNA和miRNA-155,miRNA-146a,miRNA-21表达水平,升高主动脉miRNA-126表达;主动脉miRNA-126,miRNA-155,miRNA-146a,miRNA-21与炎症因子表达具有典型相关性,其中miRNA-126为负相关,miRNA-155,miRNA-146a,miRNA-21为正相关。结果表明,丹蛭降糖胶囊对糖尿病大血管病变具有治疗效果,作用机制可能与调节miRNA-126,miRNA-155,miRNA-146a,miRNA-21水平,减少炎症因子、减轻血管炎症反应有关。

This study was aimed to investigate the mechanism of Danzhi Jiangtang Capsules(DJC)in the treatment of diabetic macrovascular disease in Goto-Kakizaki(GK)rats.The diabetic macrovascular disease rat model was induced by feeding high-fat and high-sugar combined with endothelial nitric oxide synthase(NOS)inhibitor N-nitro-L-arginine methyl ester(L-NAME)(0.1 g·L-1·d-1).According to the random array table,the model rats were randomly divided into the model group,DJC groups(1 260,630,320 mg·kg-1),atorvastatin group(105 mg·kg-1)and metformin group(10 mg·kg-1),with 12 rats in each group.The rats received gavage administration for 8 weeks.Twelve Wistar rats were selected as the normal control group.The changes of body weight,water intake,blood glucose,plasma total cholesterol(TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),interleukin(IL-1β),IL-6,tumor necrosis factor(TNF-α),nitric oxide(NO),endothelin(ET-1)were observed in these rats.Aortic tissue was taken and the pathological changes were observed by HE staining.RT-PCR was used to detect the mRNA levels of IL-1β,IL-6,and TNF-αin rat aorta.RT-PCR of the stem loop was used to detect the levels of miRNA-126,miRNA-155,miRNA-146 a,and miRNA-21 in rat plasma and aortic tissue.The canonical correlation between miRNAs and inflammatory factors was then analyzed.The results showed that DJC increased the rat body weight,lowered water intake,reduced the random blood glucose,reversed the rat aorta tissue damage,reduced serum TC,TG,LDL-C,ET-1,IL-1β,IL-6,TNF-α,as well as miRNA-155,miRNA-146 a and miRNA-21 levels in serum,elevated plasma HDL-C,NO content,reduced the aorta mRNA of IL-1β,IL-6,TNF-α,and the miRNA-155,miRNA-146 a and miRNA-21,elevated miRNA-126 expression in aorta.Aortic miRNA-126,miRNA-155,miRNA-146 a and miRNA-21 expression levels were typically correlated with the expression of inflammatory factors,among which miRNA-126 was negatively correlated,miRNA-155,miRNA-146 a and miRNA-21 were positively correlated with the factors.These results suggested that DJC had therapeutic effects on diabetic macrovascular diseases,and the mechanism of action may be related to the regulation of miRNA-126,miRNA-155,miRNA-146 a and miRNA-21 levels,as well as the reduction of inflammatory factors and vascular inflammatory response.