Affection Of Heme Oxygenase-1on The Apoptosis Of Myocardial Cells Induced By Angiotensin Ⅱ

Background and ObjectiveHeart failure is a clinical syndrome caused by structural or functional heart disease, the ability of filling or pumping blood descend, which is the end of many heart diseases such as coronary heart disease, hypertension, congenital heart disease, cardiomyopathy. With the development of medical technology and the common of Interventional treatment, the survival rate of cardiovascular diseases such as coronary heart disease and valvular heart disease has been improved greatly. Besides, as the improvement of people’s living standard, the aging population is increasing. Those leads to the number of heart failure patients also increase. Studies have confirmed that increased myocardial apoptosis is one of the important cause leading to the development of heart failure, and neuroendocrine excessive active, angiotensin Ⅱ secretion are the important mechanism of cell apoptosis, left ventricular remodeling and heart failure. If we can find the target that inhibit the myocardial cell apoptosis induced by AngⅡ, there maybe a new way to prevent the development of heart failure.Heme oxygenase (HO) widely exists in the microsomal of body’s tissues and cells, which is the key enzyme of heme degradation catalyzed. HO plays an important role in anti-oxidation, anti-inflammatory, dilating blood vessels,regulating cell proliferation and apoptosis through heme generating carbon monoxide、biliverdin、ferrous ion, which has great effections on the protection of the cardiovascular system. However, what influence HO on the myocardial cell apoptosis, there are no reports in our country.This experiment by AngⅡ inducing apoptosis of rats’myocardial cells, Add respectively HO-1inducer and inhibitor into the apoptosis of rats’myocardial cells induced by Angll. Then observe the changes of myocardial cell apoptosis, in order to investigate HO-1on myocardial cell apoptosis, provide the theoretical basis for clinical treatment and prognosis of heart failure judgment.Methods1. Culture neonatal Wistar rat’s myocardial cells primarily and Immunofluorescence identification.2. Randomly divided cells into four groups:①Control group:conventional cultivation;②Angll group:add AngⅡ;③AngⅡ+hemin group:add AngⅡ and chlorinated hemoglobin (hemin);④AngⅡ+ZnppIX group:add AngⅡ and zinc-protoporphyrin-9(ZnppIX).3. Get the totals of RNA of each group of myocardial cells, GAPDH as reference, and then use Real Time PCR to detect HO-1mRNA expression level relatively.4. Get the totals of protein of each group of myocardial cells, GAPDH as reference, then use Western blot to detect HO-1protein expression level relatively.5. Use Flow cytometry instrument and TUNEL to detect myocardial cell apoptosis rate.Results(1) The myocardial cells HO-1mRNA expression:Control group、Angll group、 AngII+hemin group and Angll+ZnppIX group HO-1mRNA expression level relatively are:1.00±0.00,3.40±1.29,8.89±2.26,1.24±0.44.①Compared with control group:AngⅡ group, AngII+hemin group and Angll+ZnppIX group were increased, Angll group, AngII+hemin group had significant differences (P<0.05); Angll+ZnppIX group hadn’t statistical difference (P>0.05);②Compared with Angll group:AngII+hemin group increased significantly, AngⅡ+ZnppIX group declined obviously, and had significance difference (P<0.05).(2) The myocardial cells HO-1protein expression:Control group、Angll group、 AngⅡ+hemin group and AngⅡ+ZnppIX group HO-1protein expression level relatively are:0.50±0.12,0.82±0.13,1.57±0.18,0.54±0.11.①Compared with control group:Angll group, AngⅡ+hemin group and Angll+ZnppⅨ group were increased, Angll group, AngⅡ+hemin group had significant differences (P<0.05); Angll+ZnppIX group hadn’t statistical difference (P>0.05);②Compared with AngⅡ group:AngII+hemin group increased significantly, AngⅡ+ZnppIX group declined obviously, and had significance difference (P<0.05).(3) Flow cytometry instrument technology measure myocardial apoptosis rate: Control group、AngⅡ group、AngⅡ+hemin group and AngⅡ+ZnppIX group myocardial cell apoptosis rate respectively are:12.1%±1.8%,26.0%±2.9%,15.2%±2.1%,39.7%±2.3%.①Compared with control group:AngⅡ3group, AngⅡ+hemin group and AngⅡ+ZnppIX group were increased, AngⅡ group, AngⅡ+ZnppIX group had significant differences (P<0.05); AngⅡ+hemin group hadn’t statistical difference (P>0.05);②Compared with AngⅡ group:AngII+hemin group decreased significantly, AngⅡ+ZnppIX group increased obviously, and both had significance difference (P<0.05).(4) TUNEL measuring apoptosis index:Control group、AngⅡ group、AngⅡ+hemin group and AngⅡ+ZnppIX group myocardial cell apoptosis rate respectively are:7.6%±2.5%,40.8%±8.2%,10.3%±2.7%,54.5%±6.3%.①Compared with control group: Angll group, AngⅡ+hemin group and Angll+ZnppIX group were increased, Angll group, Angll+ZnppⅨ group had significant differences (P<0.05); AngⅡ+hemin group hadn’t statistical difference(P>0.05);②Compared with Angll group:AngⅡ+hemin group decreased significantly, AngⅡ+ZnppⅡ group increased obviously, and both had significance difference (P<0.05).ConclusionHO-1attenuates the myocardial cells apoptosis induced by Angll.