中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE IN PRACTICE OF CRITICAL CARE MEDICINE
2014年
5期
350-353
,共4页
吴茂林%翟昌林%张亚梅%吴飞飞%张英志
吳茂林%翟昌林%張亞梅%吳飛飛%張英誌
오무림%적창림%장아매%오비비%장영지
丹参酮ⅡA%心肌缺血,急性%S100A1%细胞凋亡
丹參酮ⅡA%心肌缺血,急性%S100A1%細胞凋亡
단삼동ⅡA%심기결혈,급성%S100A1%세포조망
TanshinoneⅡA%Acute myocardial ischemia%S100A1%Apoptosis
目的：探讨丹参酮ⅡA对大鼠急性心肌缺血损伤后S100A1蛋白表达的影响。方法选择Wistar大鼠60只，按随机数字表法分为假手术组、急性心肌缺血模型组、丹参酮ⅡA预处理组。采用左冠状动脉（冠脉）前降支根部穿线结扎的方法建立大鼠急性心肌缺血模型；假手术组仅开胸于大鼠左冠脉前降支根部穿线但不结扎；丹参酮ⅡA预处理组于术前3 d腹腔注射丹参酮ⅡA注射液（剂量为1.5 mg/kg），假手术组和急性心肌缺血模型组以等体积生理盐水腹腔注射。采用原位末端缺刻标记法（TUNEL）检测心肌细胞凋亡情况；检测血清超氧化物歧化酶（SOD）、丙二醛（MDA）、肌酸激酶（CK）、乳酸脱氢酶（LDH）及S100A1蛋白水平；采用免疫组化染色和蛋白质免疫印迹试验（Western Blot）检测心肌组织S100A1蛋白的表达水平。结果与假手术组比较，心肌缺血模型组和丹参酮ⅡA预处理组心肌细胞凋亡率及MDA、CK、LDH、S100A1表达水平均明显升高，SOD活性明显降低；与心肌缺血模型组比较，丹参酮ⅡA预处理组心肌细胞凋亡率及MDA、CK、LDH、S100A1蛋白表达水平均明显降低〔细胞凋亡率：（32.1±4.2）%比（72.4±5.4）%，MDA（μmol/L）：9.1±2.2比17.3±5.2，CK（U/L）：83.3±12.2比107.5±12.4，LDH（μmol · s-1· L-1）：84.0±16.4比114.4±16.0，S100A1蛋白（μg/L）：37.6±6.0比78.4±8.6，P＜0.05或P＜0.01〕，SOD活性明显升高（kU/L：72.8±10.2比49.6±8.8，P＜0.01）。TUNEL染色结果显示，心肌缺血模型组和丹参酮ⅡA预处理组心肌细胞部分染色呈阳性（棕黄色），呈现形态不规则、与周围组织脱离和核固缩等特征，假手术组大部分细胞染色呈阴性。免疫组化结果显示，心肌缺血模型组和丹参酮ⅡA预处理组S100A1蛋白染色较深，丹参酮ⅡA预处理组阳性蛋白染色较浅。Western Blot结果显示，心肌缺血模型组S100A1蛋白表达是假手术组的2.8倍，丹参酮ⅡA预处理组较心肌缺血模型组明显降低（均P＜0.05），是假手术组的1.5倍。结论丹参酮ⅡA可能是通过抑制S100A1蛋白的表达对急性心肌缺血损伤起到保护作用，提示丹参酮ⅡA具有治疗心肌缺血的潜能。
目的：探討丹參酮ⅡA對大鼠急性心肌缺血損傷後S100A1蛋白錶達的影響。方法選擇Wistar大鼠60隻，按隨機數字錶法分為假手術組、急性心肌缺血模型組、丹參酮ⅡA預處理組。採用左冠狀動脈（冠脈）前降支根部穿線結扎的方法建立大鼠急性心肌缺血模型；假手術組僅開胸于大鼠左冠脈前降支根部穿線但不結扎；丹參酮ⅡA預處理組于術前3 d腹腔註射丹參酮ⅡA註射液（劑量為1.5 mg/kg），假手術組和急性心肌缺血模型組以等體積生理鹽水腹腔註射。採用原位末耑缺刻標記法（TUNEL）檢測心肌細胞凋亡情況；檢測血清超氧化物歧化酶（SOD）、丙二醛（MDA）、肌痠激酶（CK）、乳痠脫氫酶（LDH）及S100A1蛋白水平；採用免疫組化染色和蛋白質免疫印跡試驗（Western Blot）檢測心肌組織S100A1蛋白的錶達水平。結果與假手術組比較，心肌缺血模型組和丹參酮ⅡA預處理組心肌細胞凋亡率及MDA、CK、LDH、S100A1錶達水平均明顯升高，SOD活性明顯降低；與心肌缺血模型組比較，丹參酮ⅡA預處理組心肌細胞凋亡率及MDA、CK、LDH、S100A1蛋白錶達水平均明顯降低〔細胞凋亡率：（32.1±4.2）%比（72.4±5.4）%，MDA（μmol/L）：9.1±2.2比17.3±5.2，CK（U/L）：83.3±12.2比107.5±12.4，LDH（μmol · s-1· L-1）：84.0±16.4比114.4±16.0，S100A1蛋白（μg/L）：37.6±6.0比78.4±8.6，P＜0.05或P＜0.01〕，SOD活性明顯升高（kU/L：72.8±10.2比49.6±8.8，P＜0.01）。TUNEL染色結果顯示，心肌缺血模型組和丹參酮ⅡA預處理組心肌細胞部分染色呈暘性（棕黃色），呈現形態不規則、與週圍組織脫離和覈固縮等特徵，假手術組大部分細胞染色呈陰性。免疫組化結果顯示，心肌缺血模型組和丹參酮ⅡA預處理組S100A1蛋白染色較深，丹參酮ⅡA預處理組暘性蛋白染色較淺。Western Blot結果顯示，心肌缺血模型組S100A1蛋白錶達是假手術組的2.8倍，丹參酮ⅡA預處理組較心肌缺血模型組明顯降低（均P＜0.05），是假手術組的1.5倍。結論丹參酮ⅡA可能是通過抑製S100A1蛋白的錶達對急性心肌缺血損傷起到保護作用，提示丹參酮ⅡA具有治療心肌缺血的潛能。
목적：탐토단삼동ⅡA대대서급성심기결혈손상후S100A1단백표체적영향。방법선택Wistar대서60지，안수궤수자표법분위가수술조、급성심기결혈모형조、단삼동ⅡA예처리조。채용좌관상동맥（관맥）전강지근부천선결찰적방법건립대서급성심기결혈모형；가수술조부개흉우대서좌관맥전강지근부천선단불결찰；단삼동ⅡA예처리조우술전3 d복강주사단삼동ⅡA주사액（제량위1.5 mg/kg），가수술조화급성심기결혈모형조이등체적생리염수복강주사。채용원위말단결각표기법（TUNEL）검측심기세포조망정황；검측혈청초양화물기화매（SOD）、병이철（MDA）、기산격매（CK）、유산탈경매（LDH）급S100A1단백수평；채용면역조화염색화단백질면역인적시험（Western Blot）검측심기조직S100A1단백적표체수평。결과여가수술조비교，심기결혈모형조화단삼동ⅡA예처리조심기세포조망솔급MDA、CK、LDH、S100A1표체수평균명현승고，SOD활성명현강저；여심기결혈모형조비교，단삼동ⅡA예처리조심기세포조망솔급MDA、CK、LDH、S100A1단백표체수평균명현강저〔세포조망솔：（32.1±4.2）%비（72.4±5.4）%，MDA（μmol/L）：9.1±2.2비17.3±5.2，CK（U/L）：83.3±12.2비107.5±12.4，LDH（μmol · s-1· L-1）：84.0±16.4비114.4±16.0，S100A1단백（μg/L）：37.6±6.0비78.4±8.6，P＜0.05혹P＜0.01〕，SOD활성명현승고（kU/L：72.8±10.2비49.6±8.8，P＜0.01）。TUNEL염색결과현시，심기결혈모형조화단삼동ⅡA예처리조심기세포부분염색정양성（종황색），정현형태불규칙、여주위조직탈리화핵고축등특정，가수술조대부분세포염색정음성。면역조화결과현시，심기결혈모형조화단삼동ⅡA예처리조S100A1단백염색교심，단삼동ⅡA예처리조양성단백염색교천。Western Blot결과현시，심기결혈모형조S100A1단백표체시가수술조적2.8배，단삼동ⅡA예처리조교심기결혈모형조명현강저（균P＜0.05），시가수술조적1.5배。결론단삼동ⅡA가능시통과억제S100A1단백적표체대급성심기결혈손상기도보호작용，제시단삼동ⅡA구유치료심기결혈적잠능。
ObjectiveTo investigate the protective effect of tanshinoneⅡA on the expression of S100A1 protein after acute myocardial ischemia injury in rats.Methods Sixty Wistar rats were randomly divided into sham operation group, acute myocardial ischemia model group and tanshinoneⅡA pretreatment group by random number table. The acute myocardial ischemia model was established by thoracotomy and penetration of a thread and occlusion around the root part of the left anterior descending coronary artery, while the sham operation group was established only by thoracotomy and penetration of a thread around the root part of that artery but without occlusion; 3 days before the operation, in the tanshinoneⅡA pretreatment group, intraperitoneal injection of tanshinoneⅡA solution(at a dose of 1.5 mg/kg) was applied, while in the sham and acute myocardial ischemia groups, intraperitoneal injection of an equal volume of saline was given. Myocardial cell apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL), the levels of serum superoxide dismutase (SOD), malondialdehyde(MDA), creatine kinase(CK), lactate dehydrogenase(LDH) and S100A1 protein were examined and the level of expression of S100A1 protein in myocardial tissue was assayed by immunohistochemical staining and Western Blot.Results Compared with the sham operation group, the myocardial cell apoptosis rate, the contents of MDA, CK, LDH, S100A1 and the level of S100A1 expression in myocardial ischemia group and tanshinoneⅡA pretreated group were significantly increased, while SOD activity was decreased obviously; compared with the myocardial ischemia model group, the myocardial cell apoptosis rate, the contents of MDA, CK, LDH, S100A1 and the level of S100A1 protein expression were significantly reduced〔apoptosis rate:(32.1±4.2)% vs.(72.4±5.4)%, MDA(μmol/L): 9.1±2.2 vs. 17.3±5.2, CK(U/L): 83.3±12.2 vs. 107.5±12.4, LDH (μmol·s-1·L-1): 84.0±16.4 vs. 114.4±16.0, S100A1(μg/L): 37.6±6.0 vs. 78.4±8.6,P<0.05 orP<0.01〕, while the activity of SOD was increased markedly in tanshinoneⅡA pretreated group(kU/L:72.8±10.2 vs. 49.6±8.8,P<0.01). TUNEL staining showed that in the myocardial ischemia model group and tanshinoneⅡA pretreated group, the myocardial cells represented positive staining(brown-yellow in color), irregular in shape with nuclear pyknosis, cell detachment from the surrounding tissue and other characteristics. And in sham operation group,the staining of majority of cells was negative. The results of immunohistochemistry showed that S100A1 protein staining was relatively deep in the myocardial ischemia model group and tanshinoneⅡA pretreated group, and in the latter group, the color of S100A1 protein positive staining was not as deep as that in the former group. Western Blot showed that the S100A1 protein expression in myocardial ischemia model group was 2.8 folds of that of the sham operation group, while the S100A1 protein expression in tanshinoneⅡA pretreated group was significantly decreased compared with that of myocardial ischemia model group(bothP<0.05),which was 1.5 folds of that of the sham operation group.ConclusionTanshinoneⅡA may play a role in inhibiting the expression of S100A1 protein to protect against acute myocardial ischemia injury, suggesting that this agent have a potential effect for treatment of myocardial ischemia.
