硫化氢对大鼠糖尿病心肌病心肌间质纤维化及JAK2/STAT3通路的影响

杨萍; 庞金伟; 刘星; 吴鹏; 范忠才

doi:10.13201/j.issn.1001-1439.2016.12.021

硫化氢对大鼠糖尿病心肌病心肌间质纤维化及JAK2/STAT3通路的影响

- 1 西南医科大学(四川泸州, 646000);
- 2 西南医科大学附属医院心内科
基金项目:
四川省科学技术厅-泸州市人民政府-泸州医学院联合科研专项资金(No:0903-100020802)

详细信息

通讯作者: 范忠才,E-1mail:lzyxyxnk02@163.com

中图分类号: R542.2

收稿日期: 2016-06-10

修回日期: 2016-08-04

The influence of hydrogen sulfide on myocardial interstitial fibrosis and JAK2/STAT3signal pathway in rats diabetic cardiomyopathy

- 1 The Southwest Medical University, Luzhou, Sichuan, 646000, China;
- 2 The Affiliated Hospital of The Southwest Medical University

More Information

Corresponding author: FAN Zhongcai, E-mail: lzyxyxnk02@163.com

Received Date: 10 June 2016

Revised Date: 04 August 2016

摘要

摘要: 目的:观察硫化氢(H₂S)对糖尿病心肌病(DCM)大鼠心肌组织纤维化、JAK2/STAT3通路及炎性因子表达的影响,探讨H₂S对DCM的作用及其机制。方法:运用糖尿病饮食联合链脲佐菌素(STZ)腹腔注射法建立SD大鼠2型糖尿病模型,选取随机血糖＞16.7 mmol/L的大鼠20只随机分成DCM+0.9%氯化钠组(DCM+Saline)、DCM＋硫化氢组(DCM+NaHS),每组10只。另选20只正常大鼠随机分成正常对照组(Control+Saline)、正常对照＋硫化氢组(Control+NaHS),每组10只。从造模成功后第8周起,DCM+NaHS组和Control+NaHS组大鼠腹腔注射NaHS溶液100 μmol·kg^-1·d^-1,Control+Saline组和DCM+Saline组腹腔注射等量0.9%氯化钠。第12周末处死大鼠,取左心室心肌组织提取蛋白和制作常规石蜡切片。运用Masson染色观察大鼠心肌间质纤维化并计算心肌间质胶原容积分数(CVF);运用Western blot技术检测大鼠心肌组织Ⅰ型、Ⅲ型胶原;磷酸化JAK2(p-JAK2)、磷酸化STAT3(p-STAT3)蛋白表达水平;运用酶联免疫吸附试验(ELISA)方法检测炎性因子白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)表达水平。结果:与Control+Saline组相比,DCM+Saline组大鼠CVF明显增加,同时心肌组织中Ⅰ型、Ⅲ型胶原蛋白表达水平明显升高,p-JAK2、p-STAT3蛋白的表达水平明显升高,IL-1β、IL-6、TNF-α的表达水平也明显升高(均P＜0.05)。与DCM+Saline组相比,DCM+NAHS组大鼠CVF明显降低,心肌组织Ⅰ型、Ⅲ型胶原蛋白、p-JAK2、p-STAT3和IL-1β、IL-6、TNF-α的表达水平明显降低(均P＜0.05)。与Control+Saline组相比,Control+NaHS组各检测指标无明显差异。结论:H₂S对正常大鼠心肌间质纤维化无明显影响,但给予外源性H₂S干预可改善DCM大鼠心肌间质纤维化程度,其机制可能与下调JAK2/STAT3通路的活化有关。
- 糖尿病心肌病 /
- 心肌间质纤维化 /
- 硫化氢 /
- JAK2/STAT3 /
- 炎症
Abstract: Objective: To investigate the effects of exogenous hydrogen sulfide (H₂S) administration on the myocardial interstitial fibrosis, the activation of JAK2/STAT3 pathway and the release of inflammatory cytokines in experimental diabetic cardiomyopathy (DCM) in rats.Method: DCM models were established using streptozocin intraperitoneal injection combing with diabetic diets in young male adult Sprague-Dawley rats. NaHS was used as the donor of exogenous H₂S. 20 male type 2 diabetic rats with blood glucose higher than 16.7 mmol/L were randomly designed into DCM+Saline and DCM+NaHS group (n=10). Another 20 male rats were randomly designed into Control+Saline and Control+NaHS group (n=10). 8 weeks after the establishment of type 2 diabetes, rats in Control+NaHS and DCM+NaHS groups were injected with NaHS 100 μmol·kg^-1·d^-1 intraperitoneally. Rats in Control+Saline and DCM+Saline groups were recieved equal saline instead. All rats were sacrificed 12 weeks after the establishment of type 2 diabetes. The left heart ventricular tissues of rats were harvested. Western blotting was used to measure the expression of p-JAK2 and p-STAT3. Enzyme-1linked immunosorbent assay (ELISA) was used to quantify the release of IL-1β, IL-6 and TNF-α. Collagen volume fraction (CVF) in masson staining was calculated to explore the extent of myocardial interstitial fibrosis. Results: Compared with the Control+Saline group, the CVF, expressions of type I and type III collagen in DCM+Saline group were dramatically increased, indicating that the myocardial interstitial fibrosis was significantly higher in DCM rats. Meanwhile, the release of inflammatory cytokines including IL-1β, IL-6 and TNF-α, the expressions of p-JAK2 and p-STAT3 were also significantly upregulated in DCM rats. Fortunately, after the administration of NaHS, the CVF in DCM+NaHS group rats was dramatically decreased while the expressions of type I and type III collagen were also significantly downregulated. Moreover, the release of inflammatory cytokines including IL-1β, IL-6 and TNF-α and the expressions of p-JAK2 and p-STAT3 were also significantly downregulated.Compared with the Control+Saline group, no statistic difference of the above mentioned parameters was observed in the Control+NaHS group. Conclusion: Exogenous H₂S administration have no significant influence on the normal myocardial internal fibrosis but can significantly suppress the abnormal deposition of collagens. These results suggest that the protective effects of H₂S may be related to the modulation on JAK2/STAT3 signal pathway that worth further studying.
- diabetic cardiomyopathy /
- myocardial interstitial fibrosis /
- hydrogen sulfide /
- JAK2/STAT3 /
- inflammation

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参考文献(21)

[1]	SAELY C H,ACZEL S,MARTE T,et al.Cardiovascular complications in Type 2diabetes mellitus depend on the coronary angiographic state rather than on the diabetic state[J].Diabetologia,2004,47:145-146.
[2]	ZHOU X,AN G,LU X.Hydrogen sulfide attenuates the development of diabetic cardiomyopathy[J].Clin Sci(Lond),2015,128:325-335.
[3]	PATEL V B,MCLEAN B A,CHEN X,et al.Hydrogen sulfide:an old gas with new cardioprotective effects[J].Clin Sci(Lond),2015,128:321-323.
[4]	SZABO C.Roles of hydrogen sulfide in the pathogenesis of diabetes mellitus and its complications[J].Antioxid Redox Signal,2012,17:68-80.
[5]	HUYNH K,BERNARDO B C,MCMULLEN J R,et al.Diabetic cardiomyopathy:mechanisms and new treatment strategies targeting antioxidant signaling pathways[J].Pharmacol Ther,2014,142:375-415.
[6]	ANEJA A,TANG W H,BANSILAL S,et al.Diabetic cardiomyopathy:insights into pathogenesis,diagnostic challenges,and therapeutic options[J].Am J Med,2008,121:748-757.
[7]	赵春容,胡文星,岳志杰,等.糖尿病大鼠早期左心室心肌细胞收缩舒张功能变化[J].中国医学影像技,2014,30(2):161-166.
[8]	李倩,汪翼,孙书珍,等.糖尿病大鼠心肌胶原代谢的变化及与MMP-2/TIMP-2表达的关系[J].山东大学学报(医学版),2010,48(3):1-6.
[9]	LI C J,LV L,LI H,et al.Cardiac fibrosis and dysfunction in experimental diabetic cardiomyopathy are ameliorated by alpha-lipoic acid[J].Cardiovasc Diabetol,2012,11:73.doi:10.1186/1475-2840-11-73.
[10]	WESTERMANN D,VAN LINTHOUT S,DHAYAT S,et al.Tumor necrosis factor-alpha antagonism protects from myocardial inflammation and fibrosis in experimental diabetic cardiomyopathy[J].Basic Res Cardiol,2007,102:500-507.
[11]	肖婷,罗健,吴志雄,等.硫化氢对糖尿病大鼠心肌间质纤维化及PPARγ、NF-κB表达的影响[J].中国病理生理杂志,2015,31(4):635-639.
[12]	吴志雄,罗健,李芳,等.硫化氢对糖尿病大鼠心肌间质纤维化及NF-κB p65和TNF-α表达的影响[J].第三军医大学学报,2015,37(2):128-131.
[13]	ARUMUGGAM N,BHOWMICK N A,RUPASINGHE H P.A Review:Phytochemicals targeting JAK/STAT signaling and IDO expression in cancer[J].Phytother Res,2015,29:805-817.
[14]	CHIBA T,YAMADA M,AISO S.Targeting the JAK2/STAT3axis in Alzheimer's disease[J].Expert Opin Ther Targets,2009,13:1155-1167.
[15]	ABO-OUF H,HOOPER A W,WHITE E J,et al.Deletion of tumor necrosis factor-alpha ameliorates neurodegeneration in Sandhoff disease mice[J].Hum Mol Genet,2013,22:3960-3975.
[16]	KIM B K,TRAN H Y,SHIN E J,et al.IL-16attenuates trimethyltin-induced cognitive dysfunction via activation of JAK2/STAT3,M1 mAChR and ERK signaling network[J].Cell Signal,2013,25:1348-1360.
[17]	ZHANG M,WANG X,WANG X,et al.Oxymatrine protects against myocardial injury via inhibition of JAK2/STAT 3signaling in rat septic shock[J].Mol Med Rep,2013,7:1293-1299.
[18]	WANG X Y,ZHANG M H,YANG M L,et al.[Effect of oxymatrine on JAK2/STAT3signaling in renal tissues of rats with septic shock[J].Zhongguo Zhong Yao Za Zhi,2013,38:2696-2700.
[19]	LI T,WU S,ZHANG H,et al.Activation of nicotinic receptors inhibits TNF-α-induced production of pro-inflammatory mediators through the JAK2/STAT3signaling pathway in fibroblast-like synoviocytes[J].Inflammation,2015,38:1424-1433.
[20]	BAI K J,CHEN B C,PAI H C,et al.Thrombin-induced CCN2expression in human lung fibroblasts requires the c-Src/JAK2/STAT3pathway[J].J Leukoc Biol,2013,93:101-112.
[21]	LUAN H F,ZHAO Z B,ZHAO Q H,et al.Hydrogen sulfide postconditioning protects isolated rat hearts against ischemia and reperfusion injury mediated by the JAK2/STAT3survival pathway[J].Braz J Med Biol Res,2012,45:898-905.