姜黄素联合花色苷对睡眠剥夺合并动脉粥样硬化大鼠模型的影响

Effects of Curcumin Combined with Anthocyanin on Sleep Deprivation with Atherosclerosis in Rats

为研究姜黄素(curcumin, Cur)联合花色苷(anthocyanin, Ant)对睡眠剥夺(sleep deprivation, SD)合并动脉粥样硬化(atherosclerosis, AS)大鼠模型的保护作用及机制,将42只大鼠随机均分为正常对照组、动脉粥样硬化模型组、动脉粥样硬化+睡眠剥夺模型组、动脉粥样硬化+大平台对照组、姜黄素保护组、蓝莓花色苷保护组和姜黄素联合蓝莓花色苷保护组。除正常对照组外,其余6组高脂饲料喂养构建动脉粥样硬化模型,持续12周后,使用改良多平台水环境法对动脉粥样硬化+睡眠剥夺模型组及各保护组大鼠进行为期2周的睡眠剥夺处理。造模结束后对保护组大鼠进行对应的给药处理。随后,检测血清各项指标,并采用苏木精-伊红(hematoxylin-eosin, HE)染色观察心脏、胸主动脉变化,采用油红O染色观察胸主动脉弓部动脉粥样硬化斑块沉积情况,采用蛋白质印迹法检测大鼠心肌组织Bcl-2、胱天蛋白酶(caspase)-3和caspase-9的蛋白质表达水平。结果显示,与正常对照组相比,动脉粥样硬化模型组及动脉粥样硬化+睡眠剥夺模型组大鼠血清白细胞介素-6 (interleukin-6, IL-6)、肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)、总胆固醇(total cholesterol, TC)、甘油三酯(triglyceride, TG)水平以及空腹血糖(fasting blood glucose, FBG)水平均显著升高(P<0.05),心肌细胞和主动脉内皮细胞大量紊乱,且存在大量炎症细胞浸润,出现显著动脉粥样硬化损伤;与动脉粥样硬化模型组相比,经睡眠剥夺处理后的大鼠损伤更为严重;经姜黄素和蓝莓花色苷单独保护后的大鼠,损伤程度与模型组相比有所改善;联合给药组的心脏和主动脉无明显损伤,病变不明显。此外,模型组大鼠心肌组织中caspase-3、caspase-9的表达水平明显高于正常对照组(P<0.05), Bcl-2的表达水平明显低于正常对照组(P<0.05),经药物干预后, caspase-3、caspase-9的表达水平明显下调(P<0.05), Bcl-2的表达水平则明显上调(P<0.05)。上述结果表明,睡眠剥夺会加重动脉粥样硬化大鼠的病情发展,姜黄素和花色苷均可修护睡眠剥夺合并动脉粥样硬化大鼠心肌细胞的损伤,且二者联合使用时的修护效果更佳。

In order to study the protective effects and mechanism of curcumin(Cur)combined with anthocyanin(Ant)on the rat model of sleep deprivation(SD)and atherosclerosis(AS),42 rats were randomly divided into seven groups,including normal control group,AS model group,AS+SD model group,AS+platform control group,Cur protection group,Ant protection group,and Cur+Ant co-protection group.All the rats except those in normal control group were fed with high-fat chow for 12 weeks to construct an AS model.Then the rats in the AS+SD model group and three protection groups were treated with SD for 2 weeks using the modified multi-platform method(MMPM).At the end of modeling,the rats in the protection groups were treated with the corresponding drug administration.The serum indexes were detected,the changes in the heart and thoracic aorta were observed by hematoxylin-eosin(HE)staining,atherosclerotic plaque deposition in the arch of the thoracic aorta was observed by Oil Red O staining,and the expression levels of Bcl-2,caspase-3 and caspase-9 proteins in the myocardial tissues were detected by Western-blot.The results showed that,compared with the normal control group,both the AS model and AS+SD model groups had elevated levels of serum interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),total cholesterol(TC),triglyceride(TG),and fasting blood glucose(FBG)(P<0.05),with a large number of disordered cardiomyocytes and aortic endothelial cells,and a large number of infiltrated inflammatory cells,showing significant atherosclerotic injury.Furthermore,the AS+SD model group had a more severely damage degree than the AS model group.Compared with those in the model groups,rats in the protection groups had an alleviated damage degree after treated by either Cur or blueberry Ant,and had no significant damage and lesion in the heart and aorta when the two drugs were co-administered.In addition,the expression levels of caspase-3 and caspase-9 proteins in myocardial tissues in the model groups were significantly higher than those in the normal control group(P<0.05),and the levels of Bcl-2 proteins were significantly lower than that in the normal control group(P<0.05).Following the drug intervention,caspase-3 and caspase-9 levels were significantly downregulated(P<0.05),and Bcl-2 levels were significantly up-regulated(P<0.05)in the protection groups.The results indicated that SD would aggravate the development of AS in rats,however,both Cur and Ant could repair the damage of cardiomyocytes in rats with SD and AS,and the combination of the two drugs had a better repair effect.