西格列汀对糖尿病小鼠心肌重构和自噬的影响及其机制

doi:10.12047/j.cjap.6129.2021.073

摘要/Abstract

摘要： 目的: 探讨西格列汀对糖尿病小鼠心肌重构和自噬的影响和可能的机制。方法: 10周龄的C57小鼠腹腔注射STZ 50 mg/(kg·d),连续注射5 d,7 d测血糖浓度＞16.7 mmol/L视为糖尿病小鼠造模成功,造模成功4周后给与药物干预。本实验分四组,对照组(control, 腹腔注射等体积的缓冲液, n=10)、模型组(Streptozocin, STZ腹腔注射诱导糖尿病模型,n=8)、处理组(在模型组基础上给与西格列汀灌胃10 mg/(kg·d),n=8)、抑制剂组(在处理组的基础上给与腹腔注射Compound C (AMPK通路抑制剂,10 mg/(kg·d),n=8),对照组腹腔注射等体积缓冲液,6周后称体重,处死,取小鼠心脏并分离心室称重,计算心室/体重比,HE染色观察心肌细胞形态,Masson染色观察纤维化程度,Western blot 检测心肌脑钠肽(BNP)、转化生长因子β(TGF-β)、缝隙连接蛋白43(Cx43)、AMP依赖的蛋白激酶(AMPK)、LC3B蛋白表达。结果: 给药6周后,与对照组相比,模型组小鼠体重没有明显改变,心室/体重比明显增加(P＜0.05),苏木素-伊红(HE)染色显示细胞增大,Masson染色显示心肌间隙纤维化增多,BNP、TGF-β蛋白明显升高,Cx43、LC3B、AMPK蛋白下降(P＜0.05)。与模型组相比,西格列汀组BNP、TGF-β蛋白明显下降,Cx43、LC3B、AMPK蛋白增多(P＜0.05)。然而Compound C会抑制Cx43、LC3B、AMPK蛋白表达的上调(P＜ 0.05)。结论: 西格列汀可以改善糖尿病小鼠心肌肥厚和纤维化,并且可以通过AMPK相关通路调节Cx43和自噬。

关键词: 糖尿病, 小鼠, 心肌重构, 西格列汀, AMPK

Abstract: Objective: To investigate the effects of Sitagliptin on myocardial remodeling and autophagy in diabetic mice and its possible mechanisms. Methods: C57 mice aged ten weeks were treated with streptozocin (STZ) at the dose of 50 mg/(kg·d) by intraperitoneal injections for five consecutive days, and the level of fasting blood glucose concentration was higher than 16.7 mmol/l after seven days indicated that the diabetic model was established successfully. Mice were divided into four groups, including control group (n=10) which was intraperitoneally injected with the same volum of saline, the model group (n=8), Sitagliptin treatment group(diabetic mice were treated with Sitagliptin at the dose of 10 mg/(kg·d)by gavage, n=8) and the inhibitor group(diabetic mice were treated with Compound C, an AMPK inhibitor, at the dose of 10 mg/(kg·d) by intraperitoneal injection, n=8). After six weeks, all the mices were weighted and then put to death and the hearts were separated to caculate ventricular /body weight ratio. Hemaloxylin-Eosin (HE) staining was used to observe the cell morphology and masson staining was used to observe interstitial fibrosis. Western blot was used to test the heart protein expressions of Connexin43(Cx43), adenosine 5'-monophosphate -activated protein kinase (AMPK), brain natriuretic peptide(BNP), transforming growth factor(TGF-β) and LC3B. Results: After six weeks of treatment, compared with control group, the ventricular /body weight ratio was improved (P＜0.05), The cardiomyocyte hypertrophy and increased fibrosis were observed in the model group. The expression levels of BNP and TGF-β were increased, while the expression levels of Cx43,LC3B and AMPK were decreased significantly(P＜0.05). However, compared with model group, treatment with Sitagliptin decreased BNP, TGF-β protein levels and increased Cx43 and LC3B protein levels, while Compound C could inhibit the upregulation of Cx43, LC3B and AMPK protein (P＜0.05). Conclusion: Sitagliptin could improve cardiac hypertrophy and decrease interstitial fibrosis and AMPK-related signaling pathways was involved in the regulation of Cx43 and autophagy.

Key words: diabetes mellitus, mice, myocardial remodelling, sitagliptin, AMPK

中图分类号:

R542.2

陈奕帆, 张李君, 刘清华, 李学文. 西格列汀对糖尿病小鼠心肌重构和自噬的影响及其机制[J]. 中国应用生理学杂志, 2021, 37(5): 534-537.

CHEN Yi-fan, ZHANG Li-jun, LIU Qing-hua, LI Xue-wen. Effects of Sitagliptin on myocardial remodeling and autophagy in diabetic mice and its mechanism[J]. CJAP, 2021, 37(5): 534-537.

参考文献

[1] Cho NH, Shaw JE, Karuranga S, et al. IDF diabetes atlas: Global estimates of diabetes prevalence for 2017 and projections for 2045[J]. Diabetes Res Clin Pract, 2018, 138: 271-281.
[2] Sauve M, Ban K, Momen M, et al. Genetic deletion or pharmacological inhibition of dipeptidyl peptidase-4 improvescardiovascular outcomes after myocardial infarction in mice[J]. Diabetes, 2010, 59: 1063-1073.
[3] Al-Damrya NT, Attia HA, Al-Rasheeda NM, et al. Sitagliptin attenuates myocardial apoptosis via activating LKB-1/AMPK/Aktpathway and suppressing the activity of GSK-3β and p38α/MAPK in a ratmodel of diabetic cardiomyopathy[J]. Biomed Pharmacother, 2018, 107: 347-358.
[4] Boudina S, Abel ED. Diabetic cardiomyopathy revisited[J]. Circulation, 2007, 115: 3213-3223.
[5] Viollet B, Guigas B, Sanz Garcia N, et al. Cellular and molecular mechanisms of metformin[J]. Clin Sci(Lond), 2012, 122 (6): 253-270.
[6] Zhang H, Xiong Z, Wang J, et al. Glucagon-likepeptide-1 protects cardiomyocytes from advanced oxidation protein product-induced apoptosis via the PI3K/Akt/Bad signaling pathway[J]. Mol Med Rep, 2016, 13: 1593-1601.
[7] Yamada H, Tanaka A, Kusunose K, et al. Effect of sitagliptin on the echocardiographic parameters of left ventricular diastolic function in patients with type 2 diabetes: asubgroup analysis of the PROLOGUE study[J]. Cardiovasc Diabetol, 2017, 16(1): 63-75.
[8] 潘孙雷, 林辉, 骆杭琪, 等. 黄酒多酚对糖尿病心肌病大鼠心肌细胞凋亡的影响[J]. 中国应用生理学杂志, 2017, 33 (5): 431-435.
[9] Mizushima N, Levine B, Cuervo AM, et al. Autophagyfights disease through cellular self-digestion[J]. Nature, 2008, 451(7182): 1069-1075.
[10] Shen ZD, Chen Q, Jin TT, et al. Theaflavin 3,3′‐digallate reverses the down regulation of connexin43 and autophagy induced by high glucose via AMPK activation in cardiomyocytes[J]. J Cell Physiol, 2019, 234(10): 17999-18016.
[11] Zhou Y, Wang HY, Man FL, et al. Sitagliptin protects cardiac function by reducing nitroxidative stressand promoting autophagy in zucker diabetic fatty (ZDF) rats[J]. Cardiovascular Drugs and Therapy, 2018, 32: 541-552.
[12] Lin C, Zhang M, Zhang Y, et al. Surface peptide attenuates diabetic cardiomyopathy via AMPK-dependent autophagy[J]. Biochem Biophys Res Commun, 2017, 482(4): 665-671.
[13] Xie Z, Lau K, Eby B, et al. Improvement of cardiacfunctions by chronic metformin treatment is associated with enhanced cardiacautophagy in diabetic OVE26 mice[J]. Diabetes, 2011, 60: 1770-1778.
[14] Chen Y, Qiao X, Zhang LJ, et al. Apelin-13 regulates angiotensin ii-induced Cx43 downregulation and autophagy via the AMPK/mTOR signaling pathway in HL-1 cells[J]. Physiol Res, 2020, 69(5): 813-822.
[15] Li XT, Yu L, Gao J, et al. Apelin ameliorates high glucose-induced downregulation of connexin43 via AMPK-dependent pathway in neonatal rat cardiomyocytes[J]. Aging Dis, 2018, 9(1): 66-76.
[16] Zou MH, Xie Z. Regulation of interplay betweenautophagy and apoptosis in the diabetic heart: new role of AMPK[J]. Autophagy, 2013, 9(4): 624-625.
[17] 邵立群, 白婷婷, 赵强, 等. 黄芪注射液对缺血性心肌病大鼠心肌自噬及心肌重塑的影响[J]. 中国应用生理学杂志, 2020, 36(4): 346-349.