姜黄素调节SphK1/S1P信号通路对糖尿病合并冠心病大鼠心脏功能的影响

Impacts of Curcumin on Cardiac Function in Rats with Diabetes and Coronary Heart Disease by Regulating SphK1/S1P Signaling Pathway

糖尿病合并冠心病为糖尿病常见慢性并发症,主要原因为糖尿病导致冠状动脉粥样硬化,造成冠脉狭窄,从而引起心肌缺血、缺氧.将大鼠按照随机数字表法分为5组(12只/组):对照组、模型组、姜黄素组(200 mg/kg)、PF543组(20 mg/kg Sph K1/S1P信号通路抑制剂PF543)、姜黄素+佛波酯(PMA)组(200 mg/kg姜黄素+200μg/kg Sph K1/S1P信号通路激活剂PMA).除对照组外,其余大鼠均利用高脂高糖联合链脲佐菌素构建糖尿病大鼠模型,腹腔注射垂体后叶素复制糖尿病合并冠心病大鼠模型.超声心动图检测大鼠心脏功能;全自动生化分析仪检测大鼠血清空腹血糖(FBG)、总胆固醇(TC)、甘油三酯(TG);酶联免疫吸附(ELISA)法检测大鼠血清乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)、肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)水平;苏木精和伊红(HE)与马松(Masson)染色观察心肌组织病理损伤;原位末端标记法(TUNEL)染色观察心肌细胞凋亡;蛋白质免疫印迹(Western blot)检测心肌组织中Sph K1/S1P信号通路蛋白表达.探究姜黄素调节鞘氨醇激酶1(Sph K1)/1-磷酸鞘氨醇(S1P)信号通路对糖尿病合并冠心病大鼠心脏功能的影响.结果显示与对照组比较,模型组大鼠LVEF、FS降低,LVEDd、LVESd、FBG、TC、TG、LDH、CK-MB、胶原容积分数、TNF-α、IL-6、心肌细胞凋亡率、Sph K1、S1P表达升高(P<0.05);与模型组比较,姜黄素组、PF543组大鼠LVEF、FS升高,LVEDd、LVESd、FBG、TC、TG、LDH、CK-MB、胶原容积分数、TNF-α、IL-6、心肌细胞凋亡率、Sph K1、S1P表达降低(P<0.05);PMA显著减弱姜黄素对糖尿病合并冠心病大鼠心脏功能的改善作用(P<0.05).表明姜黄素可能通过抑制Sph K1/S1P信号通路传导减轻糖尿病合并冠心病大鼠的炎症和心肌凋亡及调节脂质代谢,进而改善其心脏功能.

Diabetes combined with coronary heart disease is a common chronic complication of diabetes.The main reason is that diabetes leads to coronary atherosclerosis,resulting in coronary stenosis,myocardial ischemia and hypoxia.In this study,Rats were randomly separated into 5 groups(12 per group)using a random number table:control group,model group,curcumin group(200 mg/kg),PF543 group(20 mg/kg SphK1/S1P signaling pathway inhibitor),and curcumin+phorbol 12-myristate 13-acetate(PMA)group(200 mg/kg curcumin+200μg/kg SphK1/S1P signaling pathway activator PMA).In addition to the control group,other rats were used to construct the diabetes rat model with high fat and high glucose combined with streptozotocin,and the rat model of diabetes combined with coronary heart disease was replicated by intraperitoneal injection of pituitrin.Ultrasound echocardiography was applied to detect cardiac function.Fully automated biochemical analyzer was applied to detect fasting blood glucose(FBG),total cholesterol(TC),and triglycerides(TG)in serum.ELISA method was applied to detect serum lactate dehydrogenase(LDH),creatine kinase isoenzyme(CK-MB),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)levels in rats.HE and Masson staining were used to observe the pathological damage of myocardial tissue.Cardiomyocyte apoptosis was observed by TUNEL staining.Western blot was applied to detect the expression of SphK1/S1P signaling pathway proteins in myocardial tissue.To explore the impacts of curcumin on cardiac function in rats with diabetes and coronary heart disease by regulating sphingosine kinase 1(SphK1)/sphingosine-1-phosphate(S1P)signaling pathway.The results showed that compared with the control group,the LVEF and FS of rats in the model group decreased,while the expression of LVEDd,LVESd,FBG,TC,TG,LDH,CK-MB,collagen volume fraction,TNF-α,IL-6,myocardial apoptosis rate,SphK1,and S1P increased(P<0.05).Compared with the model group,the LVEF and FS of rats in curcumin group and PF543 group increased,while the expression of LVEDd,LVESd,FBG,TC,TG,LDH,CK-MB,collagen volume fraction,TNF-α,IL-6,myocardial apoptosis rate,SphK1,and S1P decreased(P<0.05).PMA obviously reduced the improvement of curcumin on cardiac function in rats with diabetes and coronary heart disease(P<0.05).It is suggested that curcumin may reduce inflammation and myocardial apoptosis and regulate lipid metabolism in rats with diabetic and coronary heart disease by inhibiting SphK1/S1P signaling pathway,thereby improving their cardiac function.