背景:KATP通道调节细胞对组织缺血、缺氧的反应,介导并参与细胞或组织器官的保护作用,本课题组前期实验证实参附提取物对大鼠心肌缺血再灌注损伤有良好的保护作用,此保护效应是否与KATP通道有关,目前尚无资料报告(维普、万方数据库、medline数据库,2005-10以前).目的:观察参附提取物对在体大鼠心肌缺血再灌注损伤的保护作用,分析其与KATP通道的关系.设计:完全随机分组设计,随机对照实验.单位:大连医科大学附属第二医院麻醉科,重庆医科大学附属第二医院麻醉科.材料:清洁级成年SD大鼠24只,雌雄不拘,体质量240～320 g.参附提取物:商品名参附注射液,雅安三九药业有限公司,批号031002;格列本脲(Glibenclamide):中国药品生物制品检定所.方法:实验于2004-04/12在重庆医科大学附属第二医院麻醉实验室完成.采用在体大鼠心肌缺血再灌注损伤模型,24只大鼠完全随机分组分为4组,各6只.①假手术组:仅穿线不结扎左冠状动脉前降支,穿线后静脉注射生理盐水8 mL/kg;②缺血再灌注组:结扎前15 min静脉注射生理盐水8 mL/kg;③参附提取物组:结扎前15 min静脉注射参附提取物8 mL/kg;④参附提取物+格列本脲组:结扎前15 min静脉注射格列本脲0.33 mg/kg和参附提取物8 mL/kg.从心尖抽血6 mL,取左心室前壁分成3块,分别做匀浆、电镜和免疫组化检测.检测心肌组织中的超氧化物歧化酶活力、丙二醛含量、肿瘤坏死因子α与白细胞介素6表达、血浆中cTnI含量,观察心肌组织超微结构改变.主要观察指标:心肌组织中的超氧化物歧化酶活力、丙二醛含量、肿瘤坏死因子α与白细胞介素6表达、血浆中cTnI含量及心肌组织超微结构改变.结果:实验大鼠24只全部进入结果分析,无脱失.①缺血再灌注各组心肌超氧化物歧化酶活性较假手术组降低,坏死因子α和白细胞介素6表达升高,差异均有统计学意义(P＜0.01),缺血再灌注组丙二醛含量高于假手术组(P＜0.01);参附提取物组超氧化物歧化酶活性较缺血再灌注组升高,丙二醛含量、肿瘤坏死因子α和白细胞介素6表达降低(P＜0.01);参附提取物组和参附提取物+格列本脲组上述指标表达差异无显著性意义(P＞0.05).②与假手术组比较,缺血再灌注组、参附提取物+格列本脲组cTnI显著升高(P＜0.01);与缺血再灌注组比较,参附提取物组cTnI显著降低(P＜0.01);参附提取物组和参附提取物+格列本脲组无显著差异.③假手术组心肌细胞超微结构基本正常,线粒体轻度肿胀;缺血再灌注组核溶解,线粒体明显肿胀,大量中性粒细胞浸润;参附提取物组心肌细胞肌丝灶性溶解,线粒体肿胀,核膜完整;参附提取物+格列本脲组线粒体明显肿胀,肌丝灶性溶解,肌浆网轻度扩张,异染色边集.结论:格列本脲抑制大鼠心肌组织的KATP通道,但未消除参附对心肌缺血再灌注损伤的保护作用,KATP通道在参附提取物对心肌缺血再灌注损伤的保护作用中没有发挥重要作用.
揹景:KATP通道調節細胞對組織缺血、缺氧的反應,介導併參與細胞或組織器官的保護作用,本課題組前期實驗證實參附提取物對大鼠心肌缺血再灌註損傷有良好的保護作用,此保護效應是否與KATP通道有關,目前尚無資料報告(維普、萬方數據庫、medline數據庫,2005-10以前).目的:觀察參附提取物對在體大鼠心肌缺血再灌註損傷的保護作用,分析其與KATP通道的關繫.設計:完全隨機分組設計,隨機對照實驗.單位:大連醫科大學附屬第二醫院痳醉科,重慶醫科大學附屬第二醫院痳醉科.材料:清潔級成年SD大鼠24隻,雌雄不拘,體質量240～320 g.參附提取物:商品名參附註射液,雅安三九藥業有限公司,批號031002;格列本脲(Glibenclamide):中國藥品生物製品檢定所.方法:實驗于2004-04/12在重慶醫科大學附屬第二醫院痳醉實驗室完成.採用在體大鼠心肌缺血再灌註損傷模型,24隻大鼠完全隨機分組分為4組,各6隻.①假手術組:僅穿線不結扎左冠狀動脈前降支,穿線後靜脈註射生理鹽水8 mL/kg;②缺血再灌註組:結扎前15 min靜脈註射生理鹽水8 mL/kg;③參附提取物組:結扎前15 min靜脈註射參附提取物8 mL/kg;④參附提取物+格列本脲組:結扎前15 min靜脈註射格列本脲0.33 mg/kg和參附提取物8 mL/kg.從心尖抽血6 mL,取左心室前壁分成3塊,分彆做勻漿、電鏡和免疫組化檢測.檢測心肌組織中的超氧化物歧化酶活力、丙二醛含量、腫瘤壞死因子α與白細胞介素6錶達、血漿中cTnI含量,觀察心肌組織超微結構改變.主要觀察指標:心肌組織中的超氧化物歧化酶活力、丙二醛含量、腫瘤壞死因子α與白細胞介素6錶達、血漿中cTnI含量及心肌組織超微結構改變.結果:實驗大鼠24隻全部進入結果分析,無脫失.①缺血再灌註各組心肌超氧化物歧化酶活性較假手術組降低,壞死因子α和白細胞介素6錶達升高,差異均有統計學意義(P＜0.01),缺血再灌註組丙二醛含量高于假手術組(P＜0.01);參附提取物組超氧化物歧化酶活性較缺血再灌註組升高,丙二醛含量、腫瘤壞死因子α和白細胞介素6錶達降低(P＜0.01);參附提取物組和參附提取物+格列本脲組上述指標錶達差異無顯著性意義(P＞0.05).②與假手術組比較,缺血再灌註組、參附提取物+格列本脲組cTnI顯著升高(P＜0.01);與缺血再灌註組比較,參附提取物組cTnI顯著降低(P＜0.01);參附提取物組和參附提取物+格列本脲組無顯著差異.③假手術組心肌細胞超微結構基本正常,線粒體輕度腫脹;缺血再灌註組覈溶解,線粒體明顯腫脹,大量中性粒細胞浸潤;參附提取物組心肌細胞肌絲竈性溶解,線粒體腫脹,覈膜完整;參附提取物+格列本脲組線粒體明顯腫脹,肌絲竈性溶解,肌漿網輕度擴張,異染色邊集.結論:格列本脲抑製大鼠心肌組織的KATP通道,但未消除參附對心肌缺血再灌註損傷的保護作用,KATP通道在參附提取物對心肌缺血再灌註損傷的保護作用中沒有髮揮重要作用.
배경:KATP통도조절세포대조직결혈、결양적반응,개도병삼여세포혹조직기관적보호작용,본과제조전기실험증실삼부제취물대대서심기결혈재관주손상유량호적보호작용,차보호효응시부여KATP통도유관,목전상무자료보고(유보、만방수거고、medline수거고,2005-10이전).목적:관찰삼부제취물대재체대서심기결혈재관주손상적보호작용,분석기여KATP통도적관계.설계:완전수궤분조설계,수궤대조실험.단위:대련의과대학부속제이의원마취과,중경의과대학부속제이의원마취과.재료:청길급성년SD대서24지,자웅불구,체질량240～320 g.삼부제취물:상품명삼부주사액,아안삼구약업유한공사,비호031002;격렬본뇨(Glibenclamide):중국약품생물제품검정소.방법:실험우2004-04/12재중경의과대학부속제이의원마취실험실완성.채용재체대서심기결혈재관주손상모형,24지대서완전수궤분조분위4조,각6지.①가수술조:부천선불결찰좌관상동맥전강지,천선후정맥주사생리염수8 mL/kg;②결혈재관주조:결찰전15 min정맥주사생리염수8 mL/kg;③삼부제취물조:결찰전15 min정맥주사삼부제취물8 mL/kg;④삼부제취물+격렬본뇨조:결찰전15 min정맥주사격렬본뇨0.33 mg/kg화삼부제취물8 mL/kg.종심첨추혈6 mL,취좌심실전벽분성3괴,분별주균장、전경화면역조화검측.검측심기조직중적초양화물기화매활력、병이철함량、종류배사인자α여백세포개소6표체、혈장중cTnI함량,관찰심기조직초미결구개변.주요관찰지표:심기조직중적초양화물기화매활력、병이철함량、종류배사인자α여백세포개소6표체、혈장중cTnI함량급심기조직초미결구개변.결과:실험대서24지전부진입결과분석,무탈실.①결혈재관주각조심기초양화물기화매활성교가수술조강저,배사인자α화백세포개소6표체승고,차이균유통계학의의(P＜0.01),결혈재관주조병이철함량고우가수술조(P＜0.01);삼부제취물조초양화물기화매활성교결혈재관주조승고,병이철함량、종류배사인자α화백세포개소6표체강저(P＜0.01);삼부제취물조화삼부제취물+격렬본뇨조상술지표표체차이무현저성의의(P＞0.05).②여가수술조비교,결혈재관주조、삼부제취물+격렬본뇨조cTnI현저승고(P＜0.01);여결혈재관주조비교,삼부제취물조cTnI현저강저(P＜0.01);삼부제취물조화삼부제취물+격렬본뇨조무현저차이.③가수술조심기세포초미결구기본정상,선립체경도종창;결혈재관주조핵용해,선립체명현종창,대량중성립세포침윤;삼부제취물조심기세포기사조성용해,선립체종창,핵막완정;삼부제취물+격렬본뇨조선립체명현종창,기사조성용해,기장망경도확장,이염색변집.결론:격렬본뇨억제대서심기조직적KATP통도,단미소제삼부대심기결혈재관주손상적보호작용,KATP통도재삼부제취물대심기결혈재관주손상적보호작용중몰유발휘중요작용.
BACKGROUND: KATP channel regulates the response of cells to hypoxia and ischemia, mediates and is involved in the protection for cells or tissue organs. Our previous research confirms that Shenfu injection has good protective effect on myocardial ischemia/reperfusion injury in rats. There have been no reports on whether this protective effect is related to KATP channel in WeiPu deriodical database, WanFang database and Medline database until October 2005.OBJECTIVE: To observe the protective effect of Shenfu injection on myocardial ischemia/reperfusion injury in rats, and analyze its correlation with KATP channel.DESIGN: Completely randomized grouping design, randomized controlled trial.SETTING: Department of Anesthesiology, Second Hospital Affiliated to Dalian Medical University; Department of Anesthesiology, Second Hospital Affiliated to Chongqing Medical University.MATERIALS: Twenty-four adult SD rats of clean grade, of either gender, weighing 240 to 320 g, were employed in this trial. Shenfu injection (Sanjiu Medical & Pharmaceutical, Co., Ltd., Batch No. 031002) and glibenclamide (National Institute for the Control of Pharmaceutical and Biological Products) were employed.METHODS: This trial was carried out in the Laboratory of Anesthesiology, Second Hospital Affiliated to Chongqing Medical University from April to December 2004. Myocardial ischemia/reperfusion injury models were employed. Twenty-four rats were randomized into 4 groups, with 6 in each group: sham-operation group [putting through thread, but without ligation of left anterior descending coronary artery, 8 mL/kg normal saline, intravenous injection (i.v.)], ischemia/reperfusion group (8 mL/kg normal saline, i.v.), Shenfu injection group (8 mL/kg Shenfu injection, i.v.) and Shenfu injection + glibenclamide group (0.33 mg/kg glibenclamide and 8 mL/kg Shenfu injection, i.v.). Administration in each group was conducted 15 minutes before ligation except for that in sham-operation group (immediately after putting through thread). About 6 mL blood was taken from cardiac apex. Left ventricular anterior wall was divided into 3 parts, and then which were used for homogenate, electron microscope observation and immunohistochemical detection separatety. The superoxide dismutase (SOD) activity, malondialdehyde (MDA) level, expressions of tumor necrosis factor-α(TNF-α) and interleukin 6 (IL-6)and plasm cTnl level in myocardial tissue were detected. Ultra-structural changes of myocardial tissue were observed.MAIN OUTCOME MEASURES: SOD activity, MDA level, expressions of TNF-α and IL-6, plasm cTnI level and ultra-structural changes of myocardial tissue. RESULTS: All the 24 rats were involved in the result analysis, without deletion. ① Compared with sham-operation group,SOD activity in myocardial tissue in ischemia/reperfusion group was decreased, while the expressions of TNF-α and IL-6 were increased, with statistical difference (P ＜ 0.01). MDA level in the ischemia/reperfusion group was higher than that in the sham-operation group (P ＜ 0.01); Compared with ischemia/reperfusion group, SOD activity was increased, MDA level and expressions of TNF-α and IL-6 were all decreased in Shenfu injection group(P ＜ 0.01); There were no significant differences in above-mentioned indexes between Shenfu injection group and Shenfu injection + glibenclamide group (P ＞0.05). ②Compared with sham-operation group, the plasm cTnl level in the ischemia/reperfusion group and Shenfu injection+ glibenclamide group was significantly increased (P ＜ 0.01); Compared with ischemia/reperfusion group, plasm cTnl level in the Shenfu injection group was significantly decreased in Shenfu injection group (P ＜ 0.01); There were no signifi cant differences in plasm cTnl level between Shenfu injection group and Shenfu injection + glibenclamide group. ③ In the sham-operation group, the ultrastructure of myocardial cells was basically normal and mitochondrium swelled a little; In the ischemia/reperfusion group, karyolysis appeared, mitochondrium swelled obviously and considerable neutrophils infil trated; In the Shenfu injection group, myofilament of myocardial cells dissolved, mitochondrium swelled and nuclear mem brane was integrity; In the Shenfu injection + glibenclamide group, mitochondrium obviously swelled, myofilament present ed focus dissolving, sarcoplasmic reticulum expanded a little and allochromacy assembled in the edge of cells. CONCLUSION: Glibenclamide suppresses myocardial KATP channel, but does not eliminate the protective effect of Shenfu injection on myocardial ischemia/reperfusion injury. KTAP channel does not play an important role in the protection of Shenfu injection for myocardial ischemia/reperfusion injury.
