| Objective:Through establishing the rabbit injury model of myocardial ischemia-reperfusion, applicating methods such as histomorphology,, biochemical and molecular biology, to observe the effection of acupuncture pretreatment on the expression of endogenous protective materials such as NO, NOS, adenosine, HSPS, discussing the relationship between acupuncture pretreatment starting endogenous protection materials and the delay protective function of ischemia-reperfusion injury; and comparing if the effects of two pretreatment methods of electric acupuncture and moxibustion inducing the expression of HSPs family are consistent; providing experiment basis for the theories of "preventive treatment of disease" by acupuncture and moxibustion and "Neiguan acupoint curing the chest disease"applicated in the clinical.Methods:32New Zealand big-ear rabbits were randomly divided into4groups:control group A (sham-operation), group B(ischemia reperfusion), group C(electric acupuncture preconditioning)and group D(moxibustion pretreatment). Establishing myocardial ischemia-reperfusion injury model with the method of Ligating the anterior descending branch of the left coronary artery for40min and then reperfusing for60min. After the last bunding for48h,the rabbits in group A were opened chest, threaded and standed for40min, then pulled out the line and continue to stand for60minutes,then drawwed materials;the rabbits in group B conducted ischemia-reperfusion, then drawwed materials; rabbits in group C and D did electric acupuncture pretreatment and moxibustion pretreatment respectively,after that for48h,then conducted ischemia-reperfusion. Serum adenosine content was determined by high performance liquid chromatography; the content of serum NO, NOS was determined by ELISA method,the expression of HSPs was determined by immunohistochemical method;morphology changes of cells in left ventricular ischemia area was observed by HE staining method;ultrastructure changes of the ischemic tissue in left ventricular was observed by electron microscopy.Results:1. Morphology changes of cells in left ventricular ischemia area of MIRI rabbit was observed by HE staining method,control group A was basical normal, the myocardial cell cytoplasm was full, muscle fiber arranged neatly,interstitial had no hyperplasia, no inflammatory cells infiltrating;pathological changes of model group B was the most serious,the arrangement of myocardial cells was disordered, horizontal grain was not clear, inflammatory cells were infiltrating, muscle fiber was necrosis and dissolved; the result of group C and D were better than group B obviously,horizontal grain were more clear,mild vascular expansion could be seen in interstitial,small amount of inflammatory cells were infiltrating;2. Ultrastructure changes of the ischemic tissue in left ventricular of MIRI rabbits was observed by electron microscopy,most myocardial fibers of control group A were normal,myofilament distribution were uniform,arrangement were neat,there was only a handful of edema between muscle fibers, and mitochondria was abundant, the structure was normal;the lesions in model group B was serious, the muscle fibers shrinked,the structure was fuzzy,disordered arrangement,muscle fiber was dissolved and necrosis, interstitial was edematous,mitochondrial was edematous and cavitation,small blood vessels were expanding; the muscle tow of group C was edematous,part of myofilament was dissolved and sparse,muscle tow fracture could be seen,mitochondrial structure was normal;the myofilament of group D was regional dissolved and become loose, or had irregular contraction bands, but the mitochondria was normal. Compared with group B,the pathological changes degree of the myocardial ultrastructure of group C and D were lesser,myocardial fiber arrangement is more neat, the necrosis and dissolved scope is more limited,the mitochondria structure is more normal,this suggests that acupuncture and moxibustion preconditioning with "Neiguan acupoint" can reduce the pathological changes of myocardial ultrastructural of MIRI;3. Adenosine content comparison of each group in rabbit serum:serum adenosine content of group B(ischemia-reperfusion) decreased significantly than that of control group A with statistical difference (P＜0.05);serum adenosine content of group C and D increased significantly than that of control group B with highly statistical difference (P＜0.01), but there was no significantly difference between group C and D (P＞0.05)4. NO and NOS contents comparison of each group in rabbit serum:NO and NOS contents of group B(ischemia-reperfusion)decreased significantly than that of control group A with statistical difference (P＜0.05);NO and NOS contents of group C and D increased significantly than that of control group B with highly statistical difference (P＜0.01), but there was no significantly difference between group C and D (P＞0.05)5. HSPs expression comparison of each group in rabbit myocardial tissue:(1) compared with group A,the HSP27,70and90expression of group C and D all increased significantly (P＜0.01),the HSP27,70and90expression of group B had the rise tendency,it shows that in addition to the stress factors,acupuncture pretreatment with "Neiguan acupoint" can enhance the expression of HSP27,70and90.(2) compared with group B,the HSP27,70and90content of group D increased significantly (P＜0.01),the HSP27content of group C increased significantly (P＜0.01),HSP70and90content of group C increased (P＜0.05),it suggests that enhance the expression of HSPs by acupuncture and moxibustion pretreatment with "Neiguan acupoint" is better than that by stress response caused by surgical stimulation.(3) Compared with group C,HSP27,70and90content of group D increased (P＜0.05)Conclusion:1. Electric acupuncture pretreatment and moxibustion pretreatment have delay protective effection to myocardial tissue of MIRI rabbits,can reduce the lesions of pathological morphology and ultrastructure of myocardial tissue in rabbit;2. Electric acupuncture pretreatment and moxibustion pretreatment can increase the release of endogenous protection materials in serum of MIRI rabbits such as NO, NOS and adenosine,and induce the expression of myocardial protection protein HSPs;3. The ability of enhancing heat shock protein expression of moxibustion pretreatment is better than that of electric acupuncture pretreatment, it suggests that the expression of heat shock protein may be more sensitive to thermal stimulation. |
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