The Influence Of Myocardial Cells Apoptosis Of Cold Autologous Blood Cardioplegia To Myocardial Ischemia Reperfusion Injury In The Heart Of The Isolated Rabbits

Objective This research through the establishment of rabbit model of myocardialischemia-reperfusion injury in vitro, contrast observation of cold autologous bloodcardioplegia,HTK cardioplegia and STH cardioplegia on rabbit myocardialischemia-reperfusion injury in vitro the effect of cell apoptosis. Discuss cold autologousblood cardioplegia on myocardial ischemia reperfusion injury of myocardial protection.Method Choose the45white healthy normal rabbits, the average weight (2000-2600g)for male and female. Randomly divided into3groups: cold autologous blood cardioplegiagroup A (CABC group, n=15), B group (HTK group, n=15) and modified Thomascardioplegia group C (st. Thomas group, n=15). B and C group directly after thoracotomyin the aorta and pulmonary artery root cut out a heart; To group A after open thoracicaortic root first rapid draw blood(according to the proportion of1:4crystal mother liquorand blood configuration), drew blood into the prepared ice immediately for later use. Eachcomes out the aortic root of rabbit heart all to rinse in4℃k-h solution(to reduce theimpact on cardiac arrest and again Beating in later stage), and then quickly connection inhomemade Langendorff perfusion apparatus, perfuse15min or so to reach balanced stateadvisable; And then Each team will respectively perfuse4℃the CABC, HTK and st.Thomas cardioplegia to cardiac arrest, at the moment of the120min of cardiac arrest,perfuse k-h solution make the heart after jump and perfusion again30min or so it isadvisable to make it balance. After the experiment.And than quickly remove the heartfrom the device, shearing apex of different parts and different depth of myocardial tissue(about2×3×3cm size), immediately into the fixed in4%paraformaldehyde fixedsolution after48-72h, the paraffin embedding slice in immune histochemical method andin situ end labeling method (TUNEL method) compared to observe each group theapoptosis of myocardial cells, such as the nucleus after dyed into claybank change; Application of immune histochemical method determination of myocardial cell apoptosis(caspase3, bax and Bcl-2) number and TUNEL method to observe the number andpercentage of apoptotic cells.Result The heart of isolated rabbits infused by the three different cardioplegia,Comparedwith relevant academic research,myocardial apoptosis factors caspase-3all have increasedto some extent.1, In comparision of the Bcl-2protein expression percentage between in thethree group: Compared with CABC group and HTK group, it has no statisticalsignificance(P>0.05);Compared with CABC group and HTK group,it has the Significantstatistical difference(P<0.001);Compared with HTK group STH group has the obviousstatistical significance(P<0.01);2.In comparision of the Bax protein expression percentagebetween in the three group: Compared with CABC group and HTK group it has no statisticalsignificance(P>0.05);CABC group compared with STH group, it has the statisticallysignificant(P<0.05);HTK group compared with STH group,it has no statistical significance(P>0.05).3, In comparision of the caspase-3protein expression percentage between in thethree group: CABC group compared with HTK group,it has the statistical significance(P<0.05);CABC group compared with STH group, it has the statisticalsignificance(P<0.05);HTK group compared with STH group,it has no significant statisticalsignificance(P>0.05).4, Terminal deoxynucleotidyl transferase mediated dUTP nickendlabeling (TUNEL method):By contrast,observed the three groups of myocardial tissue cellapoptosis index (AI) percentage.Compared with CABC group and HTK group, it has nostatistical significance(P>0.05);Compared with CABC group and STH group,iit has theSignificant statistical difference(P<0.01);HTK group compared with STH group,it has theobvious statistical significance(P<0.01).Conclusion1、The influence of myocardial apoptosis exists in myocardial ischemiareperfusion injury of the heart of the isolated rabbits；2、Using cold autologous bloodcardioplegia and HTK cardioplegia to myocardial apoptosis have obvious reducecomparion with using st. Thomas cardiac cardioplegia；3、 Cold autologous bloodcardioplegia myocardial protection related to the inhibition of myocardial apoptosis during myocardial ischemia reperfusion.