Department of Basic Medicine, Sichuan Vocational College of Health and Rehabilitation, Zigong, China.
Clin Exp Pharmacol Physiol. 2021 Dec;48(12):1621-1632. doi: 10.1111/1440-1681.13569. Epub 2021 Aug 22.
This study was presented to observe the therapeutic effects of azathioprine (AZA) pretreatment on myocardial ischaemia reperfusion (I/R) damage in diabetic rats. All rats were randomly separated into control + sham operation; control +I/R; diabetes mellitus (DM) +I/R and DM +I/R + AZA groups. Diabetic rat models were established by intraperitoneally injecting 60 mg/kg streptozotocin (STZ). Diabetic rats were given 3 mg/kg AZA daily by gavage for 5 days. Then, myocardial I/R rat models were constructed. Myocardial infarction size and myocardial damage were respectively detected by TTC and H&E staining. Cardiac injury markers (CK-MB and MPO) and oxidative stress factors (SOD and MDA) were measured via ELISA. The protein expression of apoptotic markers (Caspase8, Caspase3, BAX and Bcl2), inflammatory factors (TLR4 and TNF-α) and AKT1/GSK3β in myocardial tissues was measured by western blot, immunohistochemistry or immunofluorescence. Data showed that AZA pretreatment could lessen myocardial infarction size and myocardial damage, and could down-regulate serum CK-MB, MPO, SOD and MDA levels in diabetic rats under I/R. Furthermore, AZA pretreatment decreased Caspase8, Caspase3, BAX, TLR4 and TNF-α expression, and increased Bcl2 expression in myocardial tissues of diabetic rats following I/R. Also, AZA pretreatment lowered AKT1, p-AKT1, GSK3β and p-GSK3β expression in diabetic heart after I/R. This study found that AZA may reduce myocardial injury in diabetic rats following I/R via reducing oxidative stress, cardiomyocyte apoptosis, and inflammatory response, which could be related to AKT1/GSK3β pathway inactivation.
本研究旨在观察巯嘌呤（AZA）预处理对糖尿病大鼠心肌缺血再灌注（I/R）损伤的治疗作用。所有大鼠随机分为对照组+假手术组；对照组+I/R 组；糖尿病（DM）+I/R 组和 DM+I/R+AZA 组。通过腹腔注射 60mg/kg 链脲佐菌素（STZ）建立糖尿病大鼠模型。糖尿病大鼠每日灌胃 3mg/kg AZA，共 5 天。然后构建心肌 I/R 大鼠模型。通过 TTC 和 H&E 染色分别检测心肌梗死面积和心肌损伤。通过 ELISA 测定心肌损伤标志物（CK-MB 和 MPO）和氧化应激因子（SOD 和 MDA）。通过 Western blot、免疫组化或免疫荧光法测定心肌组织中凋亡标志物（Caspase8、Caspase3、BAX 和 Bcl2）、炎症因子（TLR4 和 TNF-α）和 AKT1/GSK3β 的蛋白表达。结果显示，AZA 预处理可减少糖尿病大鼠 I/R 后的心肌梗死面积和心肌损伤，并可下调血清 CK-MB、MPO、SOD 和 MDA 水平。此外，AZA 预处理可降低糖尿病大鼠 I/R 后心肌组织中 Caspase8、Caspase3、BAX、TLR4 和 TNF-α的表达，并增加 Bcl2 的表达。同时，AZA 预处理可降低糖尿病心脏 I/R 后 AKT1、p-AKT1、GSK3β 和 p-GSK3β 的表达。本研究发现，AZA 可能通过减少氧化应激、心肌细胞凋亡和炎症反应来减轻糖尿病大鼠 I/R 后的心肌损伤，这可能与 AKT1/GSK3β 通路失活有关。
