The Preventive Effect And Mechanism Of Jinlida Granules On Cardiovascular Tissues In Sleep Deprivation Rats

Sleep is essential for the functions of the maintenance of basic life.Sleep loss,which is the state of normal sleep be not satisfied,may result in systemic metabolic disorders and cardiovascular system dysfunction.Now that the multiple mechanisms involved in sleep deprivation on cardiovascular system,such as oxidative stress,inflammation and endothelial dysfunction,autonomic nerve dysfunction,metabolic dysfunction may play a role in the pathogenesis of cardiovascular disease and development.Relatively speaking,traditional medicine have a concept of the metabolic disorder caused by insomnia in a holistic perspective.The heart and spleen lesion is the source of insomnia in the Chinese medicine thinking.The heart will be affected by spleen and stomach lesions through the meridians effect.Jinlida granules is a kind of compound hypoglycemic agents based on the theory of ?venation?in the traditional Chinese medical science.It has been shown previously that Jinlida granules may improve glucose and lipid metabolism in diabetic rats and delay the progression of chronic complications of diabetes.So can Jinlida granules also has a protective effect in metabolic and cardiovascular system dysfunction of the sleep deprivation rats?This research is mainly to study the influence of Jinlida granules on the cardiovascular system in sleep deprivation rats after 6 weeks of different doses of Jinlida granules.The experiment was divided into four parts: Part I: To observe and verify the different time points of sleep deprivation on serum metabolic oxidative stress and inflammatory response;Part II: To observe the effect of be taken different doses of Jinlida granules,through the sleep deprivation rats‘ serum and cardiac oxidative stress,inflammation and several hormones associated ventricular function;part III: To observe the effect of different doses of Jinlida granules be taken of ECG and myocardial morphological changes in sleep deprivation rats;part IV: To detect different doses of Jinlida granules on myocardial apoptosis of sleep deprivation rats,and to explore possible mechanisms.ObjectiveTo investigate the effect of the different doses of Jinlida granules on the cardiovascular system in sleep deprivation rats and to explore its possible mechanisms for the protection,and further understanding of the role and mechanism of Jinlida granules plays.MethodsUsing MMPM to establish REM sleep deprived rats model.The first study was randomly divided into CC,TC,SD1 d,SD3d and SD5 d group,the experiments of the second?third and fourth part study was randomly divided into NC,SD5 d,SD5d + JL(0.75 g / kg)and SD5 d + JH group(3.0g / kg),Each group was administered drug or placebo orally for 6 weeks.Glucose,insulin was measured by glucose oxidase – peroxidase and radioimmunoassay,the serum and myocardial SOD,MDA,GSH,MPO were measured by xanthine oxidase and TBA colorimetric,and the serum and myocardial IL-6,TNF-a,hsCRP,ANP,BNP,ET-1 change were observed by ELISA.To observe the morphological changes in myocardial tissue of sleep deprivation 5 days rats be taked different doses of Jinlida granules by ECG,HE staining and transmission electron microscopy.And also measured myocardial apoptosis by TUNEL,cardiac p-akt,Akt,Bcl-2,Bax,caspase-3 protein levels by Western blot assay.Results1.The change of weight: Compared with the normal control group,the weight of SD group were significantly lower than the control group,especially decreased in the fifth day significantly(p<0.01).Compared with the TC group,the weight gain of SD rats on the third day was reduced(p<0.05),significantly reduced in the fifth day SD rats compared with the CC and TC group(p<0.01).2.The change of FINS: Compared with CC,TC and SD1 d group,FINS of SD5 d group were significantly reduced(P<0.01).After drug intervention,compared with SD5 d group,FINS of SD5d+JH group increased(P<0.01),and SD5d+JL group increased(P<0.05).3.The change of oxidative stress indicators: In the first part of the experiment,compared with CC,TC and SD1 d,SOD vitality and GSH levels of SD3 d and SD5 d,especially SD5 d group was decreased,and MDA levels increased(P<0.01);In the Jinlida granules intervention trials,compared with SD5 d group,serum and myocardial SOD activity,GSH levels of SD5 d + JL and SD5 d + JH Group increased(P<0.01),and MDA levels decreased(P<0.01);The change of MDA levels in SD5 d + JL group was statistically significant.4.The change of Inflammatory Response: In the first part of the experiment,compared with CC and TC group,hsCRP,IL-6,TNF-? levels of SD3 d,SD5d group were significantly increased(P<0.01);in the second part of the experiment,compared with SD5 d group,serum hsCRP?serum and cardiac IL-6,TNF-? levels of SD5 d + JL,SD5 d + JH were significantly lower(P<0.01);the change of cardiac hsCRP in SD5 d + JL group was not statistically significant.5.The change of cardiovascular related factors: Compared with NC group,serum and myocardial ANP,BNP,ET-1 levels and MPO activity of SD5 d group were increased to varying degrees.Compared with the SD5 d group,the change of ANP?BNP levels in the two drug intervention groups was reduced to varying degrees.But serum and cardiac ET-1,serum MPO has no significant changes.The cardiac MPO of SD5 d + JH group decreased(P<0.05).6.ECG: Compared with NC group,myocardial ischaemia and frequent premature ventricular contractions SD5 d rats.Compared with the SD5 d group,SD5 d + JL rats restoration of sinus rhythm,regular rhythm,ST segment dropped to baseline,occasional premature ventricular contractions and atrioventricular block.SD5 d + JH group returned to the normal range,with occasional irregular rhythm.7.The morphology of cardiac tissue: H&E stained the rat ventricular tissues from the SD5 d rats show that myocardial fibers was in disarray,increased fibroblasts and the capillaries being congested;Electron microscopy showed that the cardiac tissue of SD5 d rats with fiber disorder,broken mitochondrial membrane,the ridge structure and intercalated disk structure damage,visible cytoplasmic accumulation of glycogen granules;different doses of Jinlida granules interventions have improved to varying degrees.8.Tunel Stainning in rat heart tissue: compared with NC rats,little staining was visible in NC cells,but the number of visibly stained unclei rose significantly after 5 days of SD(P <0.01);SD5d + JL and SD5 d + JH rats have improved to varying degrees.9.PI3K/Akt pathway activation and expression of apoptosis-related protein: western blotting analysis showed that,the cardiac tissue Bax,caspase3 protein levels increased to relative to the NC(P <0.01),while the expression of p-Akt,Bcl-2 protein levels were significantly lower(P <0.01).Compared with SD5 d group,SD5 d + JL,SD5 d + JH rat cardiac tissueBax,Caspase3 protein levels were decreased;and p-Akt,Bcl-2 protein expression was significantly increased(P <0.01).Conclusion1.Jinlida granules can improve Oxidation stress,inflammatory cytokines of serum and myocardial tissue of sleep deprivation rats.And the drug can also reduce the structural and ultrastructural changes in myocardial tissue pathology,to improve cardiovascular system dysfunction2.Jinlida granules may reduce myocardial apoptosis index in sleep deprivation rats.This effect may be mediated through PI3 K / Akt signaling pathway.