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微小RNA-145/TGF-β参与游泳运动对2型糖尿病大鼠血管钙化的调控

徐港港¹, 马春莲^1,2,*, 杨翼^1,2

¹武汉体育学院运动医学院运动训练监控湖北省重点实验室，武汉 430079；²武汉体育学院体质监测与慢病干预研究中心，武汉 430079

摘要

本文旨在探索游泳运动对2型糖尿病大鼠血管钙化的作用及相关分子机制。将雄性Sprague Dawley (SD)大鼠随机分为正常对照组(NC组)、糖尿病对照组(DC组)和糖尿病运动组(DE组)。DC组和DE组腹腔注射链脲佐菌素(streptozotocin, STZ)联合喂养高脂饲料进行2型糖尿病造模。NC、DC组不进行运动，DE组进行8周游泳运动。用ELISA法检测大鼠血清糖化血红蛋白A1c (glycated hemoglobin A1c, HbA1c)水平；取大鼠主动脉，用试剂盒检测血管钙含量和碱性磷酸酶(alkaline phosphatase, ALP)活性，用Von Kossa染色检测钙盐沉积，用qRT-PCR检测主动脉微小RNA-145 (microRNA-145, miR-145)表达，用Western blot检测平滑肌收缩标志分子、钙化标志分子及相关蛋白的表达。结果显示，相比NC组，DC组血糖、血清HbA1c水平、血管钙含量和ALP活性显著升高，平滑肌收缩标志分子平滑肌蛋白22α (smooth muscle protein 22α, SM22α)、α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)蛋白表达水平显著下调，钙化标志分子骨桥蛋白(osteopontin, OPN)蛋白表达水平显著上调；相较于DC组，DE组血清HbA1c水平、血管钙含量和ALP活性显著降低，SM22α、α-SMA蛋白表达水平显著上调，OPN蛋白表达水平显著下调。相比NC组，DC组miR-145-5p表达水平显著下调，转化生长因子-β (transforming growth factor-β, TGF-β)、SMAD2、ERK1/2和p-ERK1/2蛋白表达水平显著上调；相比DC组，DE组miR-145-5p表达水平显著上调，TGF-β、ERK1/2和p-ERK1/2蛋白表达水平显著下调。上述结果提示，miR-145/TGF-β参与8周游泳运动对2型糖尿病状态下糖代谢紊乱、血管平滑肌细胞表型转换和血管钙化的改善作用。

关键词： 游泳运动; 2型糖尿病; 血管钙化; 微小RNA-145

Involvement of microRNA-145/TGF-β in the regulation of swimming exercise on vascular calcification in type 2 diabetic rats

XU Gang-Gang¹, MA Chun-Lian^1,2,*, YANG Yi^1,2

¹Hubei Key Laboratory of Exercise Training and Monitoring, Department of Sports Medicine, Wuhan Sports University, Wuhan 430079, China；²Physical Fitness Monitoring and Chronic Disease Intervention Research Center, Wuhan Sports University, Wuhan 430079, China

Abstract

The present study aimed to explore the effect of swimming exercise on vascular calcification in type 2 diabetic rats and its related molecular mechanism. Male Sprague Dawley (SD) rats were randomly divided into normal control (NC), diabetes control (DC) and diabetes+exercise (DE) groups. The DC and DE groups were intraperitoneally injected with streptozotocin (STZ) and fed with high-fat diet to establish type 2 diabetes mellitus model. The NC and DC groups did not exercise, and the DE group performed swimming exercise for 8 weeks. ELISA was used to detect the serum glycated hemoglobin A1c (HbA1c) level. The aortas of rats were taken as sample. Assay kits were used to detect vascular calcium content and alkaline phosphatase (ALP) activity. Von Kossa staining was used to detect calcium deposition. qRT-PCR was used detect the expression of microRNA-145 (miR-145). Western blot was used to detect the protein expression levels of smooth muscle contraction markers, calcification marker and related proteins. The results showed that, compared with the NC group, the blood glucose, serum HbA1c level, vascular calcium content and ALP activity in the DC group were significantly increased, the protein expression levels of smooth muscle contraction markers smooth muscle protein 22α (SM22α) and α-smooth muscle actin (α-SMA) were significantly down-regulated, and the protein expression level of calcification marker osteopontin (OPN) was significantly up-regulated; Compared with the DC group, the serum HbA1c level, vascular calcium content and ALP activity in the DE group were significantly decreased, the protein expression levels of SM22α and α-SMA were significantly up-regulated, and the protein expression level of OPN was significantly down-regulated; Compared with the NC group, the expression of miR-145-5p in the DC group was significantly down-regulated, and the protein expression levels of transforming growth factor-β (TGF-β), SMAD2, ERK1/2 and p-ERK1/2 were significantly up-regulated; Compared with the DC group, the expression of miR-145-5p was significantly up-regulated in the DE group, while the expressions of TGF-β, ERK1/2 and p-ERK1/2 were significantly down- regulated. These results suggest that miR-145/TGF-β signaling is involved in the improving effects of 8-week swimming exercise on glucose metabolism disorder, vascular smooth muscle cell phenotype switching and vascular calcification in type 2 diabetes mellitus.

Key words: swimming exercise; type 2 diabetes mellitus; vascular calcification; miR-145

收稿日期：　　录用日期：

通讯作者：马春莲　　E-mail:

DOI: 10.13294/j.aps.2024.0059

引用本文：

徐港港, 马春莲, 杨翼. 微小RNA-145/TGF-β参与游泳运动对2型糖尿病大鼠血管钙化的调控[J]. 生理学报 2024; 76 (4): 526-536.

XU Gang-Gang, MA Chun-Lian, YANG Yi. Involvement of microRNA-145/TGF-β in the regulation of swimming exercise on vascular calcification in type 2 diabetic rats. Acta Physiol Sin 2024; 76 (4): 526-536 (in Chinese with English abstract).