Effects Of Puerarin Injection On Apoptosis Of Cardiac Muscle Of Exsanguine-reperfused Rat

With the prolongation of people's life-span constantly, the number of the IHD patients increases gradually and the incidence of the IHD goes up as well. As we know, the ideal therapy for AMI patients is the dissolve of thrombus. However, the subsequence damage to cardiac muscle tissue is a serious problem and concerned by many scholars in the field. To find new and safe drugs for protection of MI and damage from exsanguine-reperfused becomes a key project for medical researching workers.Puerarin injection is an extractive from dry roots of a bean faculty plant named pueta. It is a kind of single ingredient injection. Many animal and clinical researching works had demonstrated that puearin is of ability to improve cardiac metabolization, expand coronary artery, develop subaltern vessels and ameliorate the accelluar constructional changes arousing by exsanguine-reperfused. However, the study on apoptosis of cardiac cell is rarely reported. The study here try to mimic the pathological process, which the human cardiac tissue damage was made by exsanguine-reperfused from the dissolve of thrombus therapy. To examine the results by detecting the content of LOP and SOD activity in serum, the cardiac cells apoptosis and gene-related proteins Fas and Bcl2 expression were observed by light microscope to learn the relationship between cardiac cells apoptosis and exsanguine-reperfused the puerarin in exsanguine-reperfused model wasapplied the function of puerarin in exsanguine-reperfused damage, with purpose to find a safety and effective drug for survival cardiac tissues.Celiac inject 2% Pentobarbitol sodium with dose of 45mg/kg to 50 rats, open the main trachea and apply the artificial breathing, to expose the heart by removing the ribs. Set the line to the LAD for re-ligation; separated the left inner neck artery for injecting drugs later; set a cannulation for taking blood to detect LPO and SOD. The animal were divided into 5 groups with random. 1. control group: without ligated LAD, to end the experiment after 270 min. 2. exsanguine group: ligated LAD for 30min and then ending the experiment. 3. exsanguine-reperfused group: ligated LAD 30min and reperfusing 240min and then ending the experiment. The above 3 groups injected sodium Chlorice into left inner neck artery as blank control. 4. puerarin therapy A group: before ligating LAD 5min, injecting purarin with dose of lOOmg/kg bogy weight, ligated for 30min and reperfusing 240min to end experiment. 5. puerarin therapy B group: after ligated LAD 25min, injected purarin with dose of lOOmg/kg bogy weight, ligated for 30min and reperfusing 240min to end experiment. The LPO content and SOD activity were detected just before the endings of each group. The tunnel labeling, Fas and Bcl2 were detected after the the tissues stained by HE. The results were observed under the light microscope. The assay of cell apoptosis applied tunnel labeling techniques: the positive cells were stained into brown. Under the high light scope(200X), counting the cells by counting board in ten fields. The results were calculated by the number of positive cell divided total cell number. The Fas and Bcl2 protein were detected by immunohistochemistry : under light microscope with200X to count the positive cell ratio comparing the total cells in 10 random fields. The positive cells number less than 25% should be dated as negative(-); between 25%-50% should be dated as slight positive (+);between 50%-75% should be dated as positive (++); beyond 75% should be dated as strong positive (+++).The results showed puerarin therapy group A and B, compared with control group that the LPO decreased insignificantly and the activity of SOD increased sharply; puerarin therapy group A, compared with B group, the LPO content in serum and the activity of SOD without difference, which suggested the anti- oxygenation ability of puerarin, which to restrain the fatty catalase and increased endogenerated anti-catalase activity to relieve the damage of cardiac cells from exsanguine-reperfused, and maybe effective before or after exsanguine-reperfused happening. After exsanguine 30min, there was no evidence to show the apoptosis increasing, but increasing after reperfusing for 240min. Puerarin therapy group A, compared with control, the number of apoptosis cells were no diference significantly, puerarin therapy group B, compared with group A, the number of aptosis cells were decreased significantly, which suggested the injection before reperfusing can decrease the cardiac cells aptosis and the protection function of puerarin could be in stage of reperfusing. Puerarin therapy group, compared with control, the protein expression of Fas decreased while the erpression of Bcl-2 was increased sharply, which suggested could up regulated the expression of Bcl-2 and down regulated the expression of Fas to anti-aptosis in cardiac cells.We can get the conclusion from the study:...