| Objective:The SD rats were pretreated with GGA to establishis chemia/reperfusion injury model and observe the changes of myocar dialmorphology and ultrastructure?the expression of heat shock prot ein 70(HSP 70)?the changes in blood biochemical indexes(LDH,CK,CK-MB), to explore the protective effect of pretreated with GGA to ischemia/reperfusion injury in rat heat.Methods:Randomly selected 30 SD rats, male or female unrestr icted, the weight from 300 to 350 g, divided into three groups: sha m group(sham group,10 rats)?ischemia/reperfusion group(I/R group,10 rats)?GGA group(GGA group,10 rats).Sham group:3 days before surgery 2mL normal saline gavage once everyday(three times in tota l), preoperative day fasting and water deprivation, the left anterior d escending artery area wearing suture without ligation, lasting 150 m inutes and then blood sampling from carotid artery and remove the heart for further processing. I/R group:3 days before surgery 2mL normal saline gavage once everyday(three times in total),preoperativeday fasting and water deprivation, the left anterior descending arter y area wearing sutur and ligate the left anterior descending artery,30 minutes later cut the suture, with the left anterior descending arte ry blood supply recovered, lasting 120 minutes and then blood sam pling from carotid artery(the total time from the beginning of ligati on to be 150 minutes), and remove the heart for further processing.GGA group:3 days before surgery GGA be dissolved in 2mL norma l saline and then gavage(weighing SD rats in GGA group and num bered, calculated the GGA required for each rat per day according t o standard 400mg/kg/d), the next steps similar with ischemia-reperfu sion group(I/R group).Blood sampling from carotid artery and analy se the concentration changes of LDH?CK? CK—MB, perform HE staing after removal of the heart, the Western Blot test HSP70, ele ctron microscopy perspective myocardial structure.Results:1.The three groups serum LDH,CK,CK-MB concentratio n be detected by statistical analysis come out to be:the concentratio n of serum LDH,CK,CK-MB be detect were higher than sham grou p,P<0.05, there was statistically significant.The concentration of seru m LDH,CK,CK-MB detected on GGA group compared with I/R dec reased,P<0.05, there was statistically significant.2.HE staining:Sham group myocardial tissue stained uniform, ha d normal morphology, clear structure, integrated cell, arrange regularly, no bleeding and necrosis were found, showing a small amount o f inflammatory cell infiltrations, no obvious abnormalities was found.I/R group myocardial tissue arrange in disorder, muscle fiber necro sis hematomas, and severe myocardial damage were detected, had muscle fiber breakage, a large amount of inflammatory cells infiltrat ion visible. GGA group myocardial tissue arrange in disorder, but l ower than I/R group;interstitial edema between muscle fibers, some of myocardial necrosis, the degree of edema and necrosis were lo wer than I/R group, medium amount of inflammatory cells infiltrati on visible.3.Three sets of experiment myocardial tissue HSP70 gradation r atio tested by Wesrern Blot through statistical analysis come out to be:sham group gradation ratio:0.25±0.04, GGA group gradation ratio:0.69±0.04, I/R group gradation ratio: 0.47±0.08. Ischemia-reperfusion group pretreated by GGA HSP70 gradation ratio(0.69±0.04)compare d with sham group(0.25±0.04), GGA group was significantly higher,P<0.05, there was statistically significant;Ischemia-reperfusion grou p HSP70 gradation ratio( 0.47±0.08) compared with the sham gro up(0.25±0.04) was higher, P<0.05,there was statistically significant.Is chemia-reperfusion group pretreated by GGA HSP70 gradation ratio(0.69±0.04) compared with ischemia-reperfusion group(0.47±0.08),GGA group HSP70 gradation ratio was higher, P<0.05, there was statistically significant.4.Sham group myocardial tissue ultrastructure(electron microsco py) showed normal myocardial morphology, clear structure, complete d intracellular mitochondria and nucleus, inside mitochondria had hi ghly developed crest, cells arrange regularly, filaments uniform, no vesicular in cells;I/R group showed most cardiomyocytes nuclear c ondensation or rupture, muscle fibers arrange irregularly and mitoch ondrial swelling(there had serious damage within some mitochondria crest, a small number of mitochondria showed vacuoles), filaments arrange in disorder, some filaments break.GGA group myocardial ti ssue ultrastructure(electron microscopy)showed oral GGA pretreated r ats cardiac structure to be protected, had completed nucleus, mitoch ondria swelling compared with I/R group lighter, with mitochondria crest denser,no obvious vacuoles.All the results showed that GGA pretreatment of ischemia-reperfusion group(GGA group) myocardial ultrastructure damagement lower than the ischemia-reperfusion group(I/R group).Conclusions:1.GGA pretreatment group can induce myocardial t issue expression of HSP70 effectively;2.GGA pretreatment reduce myocardial ischemia reperfusion in rats the release of LDH, CK, CK-MB, reduce myocardial cell injur y;3.GGA pretreatment can improve damagement of myocardial tis sue and morphology after ischemia reperfusion, thus provide protecti ve effect to myocardial tissue. |
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