Study On The Effect Of Aralia Elata Total Saponins On Myocardial Protection In Ischemia - Reperfusion Injury By Perioperative TCM Syndromes In Patients With Emergency Coronary Intervention

Objectives1.By collecting the acute myocardial infarction patients’ four diagnostic data before and after emergency PCI, to explore the emergency PCI patients’ law of perioperative TCM syndrome, and to provide the basis for clinical diagnosis and treatment of traditional Chinese medicine.2.By establishing the rat heart perfusion in vitro model, to discuss the role and mechanism of aralosides in rat myocardial ischemia reperfusion injury, and to provide experimental basis in clinical prevention and treatment of ischemia-reperfusion.Methods1.Based on the prospective, observational study method, choose the patients conformed to the standard of acute myocardial infarction(AMI) and implemented emergency PCI surgery as the research object. Establish information collection table according to the research purpose and the related literature research conclusion.Collect the patients’ basic information, four diagnostic informations and establish the database of traditional Chinese medicine. Analyse the data by using spss18.0 clustering analysis method. Explore the acute myocardial infarction patients’ traditional Chinese medicine syndrome before and after emergency PCI.2.60 male rats are randomly divided into control group, model group, DZ group, 5-HD group, aralosides group, aralosides+5-HD group, each group ten rats. Using Lan gendroff isolated heart retrograde perfusion system to built isolated heart retrograde pe rfusion models in rats, HE staining to observe the myocardial tissue, western-blot semi-quantitative to determinate the level of cytochromeC, JC-1 method to determinate me mbrane potential, immunohistochemical method to detect Caspase-3, Bax, Bcl-2 express ion,enzyme-linked immunoassay to detect the content of mitochondrial ATP, mitochondr ial Ca2+, Myocardial tissue MDA,SOD, ROS,GSH-PX.Results1.The AMI patients’male to female ratio is 4.09:1, which has obvious difference. The AMI patients’age gathers in 40 to 70 years old, the patients are mostly recent onset, Front wall and lower wall AMI account for larger proportion. Most of the criminal blood vessels is the descending branch, followed by the right and the swing. Twenty-six patients with ischemia-reperfusion injury, accounts for 23.2%. AMI patients with combined diseases are in the majority of high blood pressure and diabetes. Research the four diagnostic informations by the clustering analysis. The result puts emergency PCI preoperative into five types, emergency PCI postoperative into four types. The traditional Chinese medicine syndrome types in emergency PCI preoperative are phlegm and stagnated blood syndrome, Qi deficiency and blood stasis syndrome, heart-blood stasis syndrome, Qi Yin deficiency syndrome, Qi stagnation and blood stasis syndrome. The traditional Chinese medicine syndrome types in emergency PCI postoperative are Qi deficiency and blood stasis syndrome, Qi deficiency and blood stasis and turbid phlegm syndrome, Qi Yin deficiency syndrome, Qi stagnation and blood stasis syndrome. The large proportion of emergency PCI preoperative is phlegm and stagnated blood syndrome, Qi deficiency and blood stasis syndrome. Syndrome of intermingled deficiency and excess is in the majority of emergency PCI postoperative patients. Qi deficiency and blood stasis syndrome is the most, followed by Qi deficiency and blood stasis and turbid phlegm syndrome. The proportion of Qi deficiency and blood stasis syndrome in emergency PCI preoperative is 42.31%, the proportion of Qi deficiency and blood stasis syndrome in emergency PCI postoperative is 61.54%.2.①HE dyeing:The control group:Myocardial cell horizontal grain is clear, the nucleus is in the middle, and orderly arranged rules, without cells degeneration and thanatosis, interstitial blood vessels without expansion, without Inflammatory cells infiltration. Model group:Myocardial cell moderate swelling degeneration, local cell sarcoplasm dissolved cavity change, disordered arrangement of cells and local myocardial tissue fibrosis, interstitial loose, much quantity of inflammatory cell infiltration, angiectasis hyperemia.DZ group:Myocardial cell horizontal grain clarity, the nucleus is in the middle, regularlyand orderly, mild swelling of cell degeneration, interstitial osteoporosis, a small amount of inflammatory cells infiltration. 5-HD group:Myocardial cell moderate swelling degeneration, local cell sarcoplasm dissolved cavity change, myocardial disorder, focal hyperplasia of fibrous tissue, muscle is island, interstitial loose, much quantity of inflammatory cell infiltration, angiectasis hyperemia. Aralosides group:Myocardial cell horizontal grain clarity, nuclear center, regular and orderly, mild swelling of cell degeneration, increase muscle bundle spacing, interstitial osteoporosis, a small amount of inflammatory cells infiltration. Aralosides+5-HD group:Mild swelling of myocardial cell degeneration, local myocardial cell rupture, cells arranged disorderly, increase myocardial beam spacing, interstitial osteoporosis, more quantity of inflammatory cell infiltration, angiectasis hyperemia. ②The level of cytochrome C:Compared with the control group, the relative expression of mitochondrial cytochrome C is obviously decreased in model group (p<0.05). Compared with the model group, the relative expression of mitochondrial cytochrome C is significantly increased (P<0.01) in DZ group and aralosides group. There is no obvious difference between the DZ group and aralosides group in the relative expression of mitochondrial cytochrome C(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the relative expression of mitochondrial cytochrome C(P>0.05). ③The mitochondrial membrane potential:Compared with the control group, the mitochondrial membrane potential is obviously decreased in model group(P<0.01). Compared with the model group, the mitochondrial membrane potential is significantly increased in DZ group and aralosides group (P<0.01). Compared with the model group, the DZ group and the aralosides group, the mitochondrial membrane potential is obviously decreased in 5-HD group and aralosides+5-HD group(P<0.01). There is no obvious difference between the DZ group and aralosides group in the mitochondrial membrane potential(P>0.05).There is no obvious difference between the 5-HD group and aralosides+5-HD group in the mitochondrial membrane potential(P>0.05).④Caspase-3, Bax, Bcl-2 expression:Compared with the control group, the Caspase-3 expression is significantly increased in the model group, the DZ group, the 5-HD group and the aralosides+5-HD group(P<0.01), is increased in the aralosides group(P<0.05). Compared with the model group, the Caspase-3 expression is increased in the 5-HD group(P<0.05), is decreased in the DZ group and the aralosides group(P<0.05).There is no obvious difference between the aralosides+5-HD group and the model group even though the former is higher(P>0.05). There is no obvious difference between the DZ group and aralosides group in the Caspase-3 expression(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the Caspase-3 expression(P>0.05). Compared with the control group, the Bax expression is increased in the model group(P<0.05),is obviously increased in the 5-HD group and aralosides+5-HD group(P<0.01),no difference in the DZ group and the aralosides group(P>0.05). Compared with the model group, the Bax expression is decreased in the in the DZ group and the aralosides group(P<0.05),is increased in the 5-HD group(P<0.05), no difference in the aralosides+5-HD group(P>0.05). There is no obvious difference between the DZ group and aralosides group in the Bax expression(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the Bax expression(P>0.05). Compared with the control group, the Bcl-2 expression is decreased in the model group(P<0.05),is obviously decreased in the 5-HD group and aralosides+5-HD group(P<0.01),no difference in the the DZ group and the aralosides group(P>0.05). Compared with the model group, the Bcl-2 expression is increased in the in the DZ group and the aralosides group(P<0.05),is decreased in the 5-HD group(P<0.05), no difference in the aralosides+5-HD group(P>0.05). There is no obvious difference between the DZ group and aralosides group in the Bcl-2 expression(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the Bcl-2 expression(P>0.05).⑤Content of mitochondrial ATP:Compared with the control group, the content of mitochondrial ATP is obviously decreased in each group(P<0.01). Compared with the model group, the content of mitochondrial ATP is obviously increased in the DZ group and the aralosides group(P<0.01),is obviously decreased in the 5-HD group(P<0.01),is decreased in the aralosides+5-HD group(P<0.05). Compared with the DZ group, the content of mitochondrial ATP is obviously increased in the aralosides group(P<0.01). Compared with the 5-HD group, the content of mitochondrial ATP increased in the aralosides+5-HD group(P<0.05).⑥Content of mitochondrial Ca2+:Compared with the control group, the content of mitochondrial Ca2+ is obviously increased in each group(P<0.01). Compared with the model group, the content of mitochondrial Ca2+ is obviously decreased in the DZ group and the aralosides group(P<0.01),is obviously increased in the 5-HD group and the aralosides+5-HD group (P<0.01). Compared with the DZ group, the content of mitochondrial Ca2+ is decreased in the aralosides group(P<0.05). Compared with the 5-HD group, the content of mitochondrial Ca2+ is decreased in the aralosides+5-HD group(P<0.05).⑦Content of MDA、 SOD、ROS、GSH-PX: Compared with the control group, the content of MDA is obviously increased in each group(P<0.01). Compared with the model group, the content of MDA is obviously decreased in the DZ group and the aralosides group(P<0.01), is obviously increased in the 5-HD group and the aralosides+5-HD group(P<0.01). There is no obvious difference between the DZ group and aralosides group in the content of MDA(P>0.05). Compared with the 5-HD group, the content of MDA is obviously decreased in the aralosides+5-HD group(P<0.01). Compared with the control group, the content of SOD is obviously decreased in the model group, the aralosides group, the 5-HD group and the aralosides+5-HD group(P<0.01),is decreased in the DZ group(P<0.05). Compared with the model group, the content of SOD is obviously increased in the DZ group and the aralosides group(P<0.01),is obviously decreased in the 5-HD group(P<0.01), no difference in the aralosides+5-HD group(P>0.05). There is no obvious difference between the DZ group and aralosides group in the content of SOD(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the content of SOD(P>0.05). Compared with the control group, the content of ROS is obviously increased in the model group, the DZ group, the 5-HD group and the aralosides+5-HD group(P<0.01),is increased in the aralosides group(P<0.05). Compared with the model group, the content of ROS is obviously decreased in the DZ group and the aralosides group(P<0.01), is obviously increased in the 5-HD group and the aralosides+5-HD group (P<0.01). There is no obvious difference between the DZ group and aralosides group in the content of ROS(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the content of ROS(P>0.05). Compared with the control group, the content of GSH-PX is obviously decreased in the model group, the DZ group, the 5-HD group and the aralosides+5-HD group(P<0.01),is decreased in the aralosides group(P<0.05). Compared with the model group, the content of GSH-PX is obviously increased in the DZ group and the aralosides group(P<0.01),is obviously decreased in the 5-HD group(P<0.01), no difference in the aralosides+5-HD group(P>0.05). There is no obvious difference between the DZ group and aralosides group in the content of GSH-PX(P>0.05). There is no obvious difference between the 5-HD group and aralosides+5-HD group in the content of GSH-PX(P>0.05).Conclusions1.Before the emergency PCI, the patients’real symptoms are the principal contradictions. The chest pain is the outstanding performance of all the syndromes, it is all because of the blocking of blood stasis. After the emergency PCI, most patients with symptoms such as chest pain, chest tightness are improved, the sign of weakness is performed, but the real symptom is always blood stasis. After opening the criminal vascular, patients with Qi deficiency and blood stasis syndrome are more likely to be ischemia-reperfusion injured.2.After MIRI, the myocardial tissue has obviously pathological changes. The myo cardial cell swelling, the cytoplasm dissolved, myocardial tissue fibrosis, inflammatory cells infiltration, blood cells expansion. The relative expression of mitochondrial cytoch rome C, the mitochondrial membrane potential, the content of mitochondrial ATP decre ase, the content of mitochondrial Ca2+ increases, cell apoptosis material increases. The oxidative stress and resistance to oxidative stress disorder. The aralosides can open mitoKATP, stabilize the mitochondrial membrane potential, reduce the mitochondrial me mbrane permeability, improve energy metabolism, promote the synthesis of mitochondri al high-energy phosphate compound, promote the mitochondrial respiratory function rec overy, reduce calcium overload, inhibit oxidative stress reaction, inhibit myocardial apo ptosis, protect the ischemia-reperfusion injury of myocardial cells, but the protection ro le of the aralosides can be decreased by mitoKATP inhibitor 5-HD.