辛伐他汀对大鼠糖尿病所致心肌细胞凋亡的影响及其机制

doi:10.12047/j.cjap.5653.2018.096

中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (5): 422-426.doi: 10.12047/j.cjap.5653.2018.096

辛伐他汀对大鼠糖尿病所致心肌细胞凋亡的影响及其机制

李凡璐¹, 宛欣¹, 王茜¹, 刘鑫¹, 吴亚俐¹, 陈还珍², 崔香丽¹

1. 山西医科大学生理系省部共建细胞生理学重点实验室, 太原 030001;
2. 山西医科大学第一医院心血管内科, 太原 030001

收稿日期:2017-11-28 修回日期:2018-09-25 出版日期:2018-09-28 发布日期:2019-02-21
通讯作者: 崔香丽,Tel:0351-4135329;E-mail:cuixlcxl@sina.com E-mail:cuixlcxl@sina.com
基金资助:
国家自然科学基金（81272695）；山西省‘1331工程’重点学科建设计划经费资助；山西省自然科学基金（20160D011107）

Simvastatin prevented myocardium of diabetes rats from apoptosis through inhibition of oxidative stress

LI Fan-lu¹, WAN Xin¹, WANG Xi¹, LIU Xin¹, WU Ya-li¹, CHEN Huan-zhen², CUI Xiang-li¹

1. Department of Physiology, Shanxi Medical University, Taiyuan 030001;
2. Department of Cardiovascular Internal Medicine, First Hospital of Shanxi Medical University, Taiyuan 030001, China

Received:2017-11-28 Revised:2018-09-25 Online:2018-09-28 Published:2019-02-21
Supported by:
国家自然科学基金（81272695）；山西省‘1331工程’重点学科建设计划经费资助；山西省自然科学基金（20160D011107）

摘要/Abstract

摘要： 目的：探讨辛伐他汀对糖尿病所致心肌损伤的保护作用及可能机制。方法：24只SD大鼠（180~220） g随机分为对照组（control组，n=8）和模型组（n=16），模型组给予链脲佐菌素（STZ）腹腔注射建立糖尿病模型，然后随机将模型组分为糖尿病组（DM组，n=8）和糖尿病+辛伐他汀组（DM+S组，n=8），DM+S组给予辛伐他汀40 mg/（kg·d）灌胃四周，其余两组给予的等量生理盐水。实验结束后，取心脏，分光光度测定法测定大鼠心肌组织脂质过氧化物丙二醛（MDA）含量、超氧化物歧化酶（SOD）的活性，HE染色观察大鼠心肌病理学改变，TUNEL法检测大鼠心肌细胞凋亡，免疫组织化学法测定大鼠p53的表达；Western blot检测心肌组织p53，p53-phospho-serine15，Bax，Bcl-2等蛋白的表达。结果：①与对照组比较，DM组大鼠心肌组织MDA含量显著升高（P<0.01），SOD活性明显下降（P<0.01），与DM组比较，DM+S组大鼠SOD活性显著增加（P<0.01），MDA含量显著下降（P<0.01）。②HE染色结果表明，与对照组比较，DM组大鼠心肌细胞排列紊乱，结构不清晰，有大量炎性细胞浸润；与DM组比较，DM+S组的心肌细胞结构明显改善。③ TUNEL细胞凋亡检测结果表明，与control组相比，DM组心肌凋亡细胞阳性率显著增加（P<0.01），给予辛伐他汀后细胞凋亡明显减少（P<0.01）；④免疫组织学检测结果显示，与对照组比较，DM组p53表达量显著增加，且p53在细胞质和细胞核中均有表达（P<0.01）；与DM组比较，DM+S组p53表达量明显下降，p53在细胞质和核中均有表达，但核内表达量减少（P<0.01）。⑤ Western blot检测结果表明，与对照组比较，DM组大鼠p53，p53-Phospho-Serine15，Bax表达量显著升高（P<0.01），Bcl-2表达量减少（P<0.01）；与DM组比较，DM+S组p53（P<0.01），p53-Phospho-Serine15（P<0.01），Bax （P<0.05）表达量显著下降，Bcl-2（P< 0.05）表达量显著增加。结论：辛伐他汀通过改善心肌结构异常、抑制氧化应激和心肌细胞凋亡对糖尿病导致的心肌损伤起到保护作用，其机制与p53介导的细胞凋亡通路相关蛋白的调控有关。

关键词: SD大鼠, 糖尿病, 心脏, 辛伐他汀, 凋亡, 氧化应激

Abstract: Objective: To investigate the protective effects and the possible mechanisms of simvastatin on myocardial injury induced by diabetes.Methods: Twenty-four SD rats (180~220)g were randomly divided into control group (control, n=8) and modeled groups(n=16), the modeled groups were injected with streptozotocin intraperitoneally to induce diabetes. Then the modeled rats were randomly divided into diabetes mellitus group (DM group, n=8) and diabetes mellitus + simvastatin group (DM+S group, n=8). Rats in DM+S group were treated with simvastatin at the dose of 40 mg/(kg·d)by gavage for 4 weeks, and the other two groups were treated with the same amount of saline. At the end of experiments, the heart tissues were collected for further observation. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in heart tissues were measured by spectrophotometry; HE staining of rat heart slides was used to observe the pathological changes; TUNEL assay was used to determine the apoptosis index of myocardial cells in each groups; The distribution of p53 in the heart tissues was evaluated by immunohistochemistry; Western blot was used to detect the expressions of p53, p53-phospho-serine 15, Bax and Bcl-2 in the heart tissues. Results: ①Compared with control group, the content of malondialdehyde (MDA) was increased while the activity of superoxide dismutase (SOD) was decreased significantly in DM group (P<0.01). After simvastatin administration, the activity of SOD was increased and the content of MDA was decreased significantly (P<0.01). ② HE staining results showed that the myocardial cells in the DM group were disorganized, with unclear morphological structure and a large number of inflammatory cells infiltration. Compared with DM group, the myocardial morphology in DM+S group was improved significantly. ③TUNEL staining results showed that the apoptosis index of myocardial cells in DM group was increased significantly compared with that of control group, and the apoptosis index was decreased significantly after the treatment of simvastatin (P<0.01).④ Immunohistochemistry showed that compared with control group,the expression of p53 in DM group was increased significantly, and was expressed in both cytoplasm and nucleus, while the expression of p53 in DM+S group was decreased and the expression of p53 in nucleus was decreased significantly (P<0.01). ⑤ The results of Western blot showed that the expression levels of p53, p53-phospho-serine15 and Bax were higher than those in control group, and the expression of Bcl-2 was lower than that in control group (P<0.01). After simvastatin administration, the expression levels of p53,p53-phospho-serine 15 (P<0.01) and Bax were decreased significantly (P<0.05) and the expression of Bcl-2 was increased (P<0.05). Conclusion: Simvastatin exerted protective effects on myocardial injury caused by diabetes through improving the abnormal morphological changes of diabetic myocardium, alleviating oxidative stress and inhibiting apoptosis of myocardial cells. The mechanism is related to the regulation of apoptosis pathway mediated by p53.

Key words: SD rats, diabetes mellitus, heart, simvastatin, apoptosis, oxidative stress

李凡璐, 宛欣, 王茜, 刘鑫, 吴亚俐, 陈还珍, 崔香丽. 辛伐他汀对大鼠糖尿病所致心肌细胞凋亡的影响及其机制[J]. 中国应用生理学杂志, 2018, 34(5): 422-426.

LI Fan-lu, WAN Xin, WANG Xi, LIU Xin, WU Ya-li, CHEN Huan-zhen, CUI Xiang-li. Simvastatin prevented myocardium of diabetes rats from apoptosis through inhibition of oxidative stress[J]. CJAP, 2018, 34(5): 422-426.

参考文献

[1] Chavali V, Tyagi SC, Mishra PK. Predictors and prevention of diabetic cardiomyopathy[J]. Diabetes Metab Syndr Obes, 2013, 6:151-160.
[2] Zhao MX, Zhou B, Ling L, et al. Salusin-beta contributes to oxidative stress and inflammation in diabetic cardiomyopathy[J]. Cell Death Dis, 2017, 8(3):e2690.
[3] Kumar S, Prasad S, Sitasawad SL. Multiple antioxidants improve cardiac complications and inhibit cardiac cell death in streptozotocin-induced diabetic rats[J]. PLoS One, 2013, 8(7):e67009.
[4] Crisby M. Modulation of the inflammatory process by statins[J]. Timely Top Med Cardiovasc Dis, 2005, 9:E3.
[5] Kumai T, Matsumoto N, Koitabashi Y, et al. Pleiotropic effects of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors:candidate mechanisms for anti-lipid deposition in blood vessels[J]. Curr Med Chem Cardiovasc Hematol Agents, 2005, 3(3):195-201.
[6] 王妍妍, 陈还珍, 李艳, 等. 辛伐他汀对糖尿病大鼠低血糖时血清炎症因子及血管内皮的影响[J]. 中西医结合心脑血管病杂志, 2013, 11(7):851-852.
[7] Li H, Zhao QQ, Liu R, et al. Protective effect and potential mechanism of simvastatin on myocardial injury induced by diabetes with hypoglycemia[J]. Exp Clin Endocrinol Diabetes, 2018, 126(3):148-161.
[8] 许良中, 杨文涛. 免疫组织化学反应结果的判断标准[J]. 中国癌症杂志, 1996(4):229-231.
[9] Huynh K, Bernardo BC, McMullen JR, et al. Diabetic cardiomyopathy:mechanisms and new treatment strategies targeting antioxidant signaling pathways[J]. Pharmacol Ther, 2014, 142(3):375-415.
[10] Chen YH, Feng B, Chen ZW. Statins for primary prevention of cardiovascular and cerebrovascular events in diabetic patients without established cardiovascular diseases:a meta-analysis[J]. Exp Clin Endocrinol Diabetes, 2012, 120(2):116-120.
[11] Olafsdottir E, Aspelund T, Sigurdsson G, et al. Similar decline in mortality rate of older persons with and without type 2 diabetes between 1993 and 2004 the Icelandic population-based Reykjavik and AGES-Reykjavik cohort studies[J]. BMC Public Health, 2013, 13:36.
[12] Silva S, Lourenço P, Paulo C, et al. Statin-induced low cholesterol is not associated with poor outcome in chronic heart failure[J]. J Cardiovasc Pharmcol Ther, 2012, 17(3):284-290.
[13] Koh GC, Vlaar AP, Hofstra JJ, et al. In the critically ill patient, diabetes predicts mortality independent of statin therapy but is not associated with acute lung injury:a cohort study[J]. Crit Care Med, 2012, 40(6):1835-1843.
[14] Janda S, Young A, Fitzgerald JM, et al. The effect of statins on mortality from severe infections and sepsis:a systematic review and meta-analysis[J]. J Crit Care, 2010, 25(4):656.
[15] Wemmelund H, H￠gh A, Hundborg HH, et al. Statin use and rupture of abdominal aortic aneurysm[J]. Br J Surg, 2014, 101(8):966-975.
[16] Yu Y, Ohmori K, Chen Y, et al. Effects of pravastatin on progression of glucose intolerance and cardiovascular remodeling in a type Ⅱ diabetes model[J]. J Am Coll Cardiol, 2004, 44(4):904-913.
[17] Ozaki T, Nakagawara A. p53:the attractive tumor suppressor in the cancer research field[J]. J Biomed Biotechnol, 2011, 2011:603925.
[18] Appella E, Anderson CW. Post-translational modifications and activation of p53 by genotoxic stresses[J]. Eur J Biochem, 2001, 268(10):2764-2772.
[19] Chen YW, Chenier I, Chang SY, et al. High glucose promotes nascent nephron apoptosis via NF-κB and p53 pathways[J]. Am J Physiol-Renal, 2011, 300(1):F147-156.
[20] Huang J, Li X, Li M, et al. Mitochondria-targeted antioxidant peptide SS31 protects the retinas of diabetic rats[J]. Curr Mol Med, 2013, 13(6):935-945.

辛伐他汀对大鼠糖尿病所致心肌细胞凋亡的影响及其机制

Simvastatin prevented myocardium of diabetes rats from apoptosis through inhibition of oxidative stress

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