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
2015年
1期
33-37
,共5页
唐关敏%翟昌林%胡惠林%徐文博
唐關敏%翟昌林%鬍惠林%徐文博
당관민%적창림%호혜림%서문박
缺血/再灌注损伤,心肌%针刺预处理%高迁移率族蛋白B1
缺血/再灌註損傷,心肌%針刺預處理%高遷移率族蛋白B1
결혈/재관주손상,심기%침자예처리%고천이솔족단백B1
Ischemia/reperfusion injury,myocardial%Electroacupuncture pretreatment%High mobility group box 1
目的：探讨针灸预处理对大鼠心肌缺血/再灌注（I/R）后的保护作用以及对高迁移率族蛋白B1（HMGB1）表达的影响。方法选择60只Wistar大鼠，按随机数字表法分为假手术组、心肌I/R模型组、针灸预处理组，每组20只。采用结扎冠状动脉左室支左心耳下缘约0.5 cm处阻断血流10 min后再灌注1 h制备I/R损伤模型；假手术组仅穿线不结扎；针灸预处理组于I/R前7 d给予每日1次电针内关穴20 min，连续治疗7 d。采用苏木素-伊红（HE）染色，光镜下观察心肌组织病理学变化，半定量积分法计算3组心肌组织病理学评分；采用酶联免疫吸附试验（ELISA）检测血浆HMGB1、肿瘤坏死因子-α（TNF-α）、心肌肌钙蛋白T（cTnT）的含量，逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹试验（Western Blot）检测心肌组织HMGB1、单核细胞趋化蛋白-1（MCP-1）、TNF-α的mRNA及蛋白表达水平。结果光镜下可见心肌I/R模型组心肌纤维部分断裂，心肌细胞大片状坏死，边界不清，细胞出现浓缩、破裂、溶解、甚至消失，间质水肿并伴大量炎性细胞浸润；针灸预处理组上述表现较心肌I/R模型组明显减轻。与假手术组比较，心肌I/R模型组HMGB1、TNF-α、cTnT含量和组织病理学评分均明显升高〔HMGB1（μg/L）：9.64±1.16比2.15±.031，TNF-α（μg/L）：91±22比19±5， cTnT（μg/L）：1.50±0.35比0.07±0.03，组织病理学评分（分）：2.5±0.3比0.0±0.0，均P＜0.01〕，HMGB1、MCP-1、TNF-αmRNA和蛋白表达均明显升高（HMGB1 mRNA：1.42±0.16比0.02±0.00，MCP-1 mRNA：0.46±0.06比0.01±0.00，TNF-αmRNA：0.75±0.04比0.03±0.00；HMGB1蛋白：1.08±0.01比0.02±0.01， MCP-1蛋白：0.92±0.03比0.40±0.01，TNF-α蛋白：1.10±0.02比0.35±0.01，P＜0.05或P＜0.01）；与心肌I/R模型组比较，针灸预处理组HMGB1（6.58±0.73）、TNF-α（63±19）、cTnT（1.15±0.31）含量均明显降低（均P＜0.01），HMGB1、MCP-1、TNF-αmRNA和蛋白表达明显降低（mRNA表达分别为0.74±0.12、0.18±0.02、0.10±0.03，蛋白表达分别为0.40±0.01、0.36±0.02、0.50±0.02，均P＜0.05），组织病理学评分（1.2±1.0）明显降低（P＜0.01）。结论针灸预处理可减轻大鼠心肌I/R损伤，其机制可能与减轻HMGB1介导的晚期炎症反应有关。
目的：探討針灸預處理對大鼠心肌缺血/再灌註（I/R）後的保護作用以及對高遷移率族蛋白B1（HMGB1）錶達的影響。方法選擇60隻Wistar大鼠，按隨機數字錶法分為假手術組、心肌I/R模型組、針灸預處理組，每組20隻。採用結扎冠狀動脈左室支左心耳下緣約0.5 cm處阻斷血流10 min後再灌註1 h製備I/R損傷模型；假手術組僅穿線不結扎；針灸預處理組于I/R前7 d給予每日1次電針內關穴20 min，連續治療7 d。採用囌木素-伊紅（HE）染色，光鏡下觀察心肌組織病理學變化，半定量積分法計算3組心肌組織病理學評分；採用酶聯免疫吸附試驗（ELISA）檢測血漿HMGB1、腫瘤壞死因子-α（TNF-α）、心肌肌鈣蛋白T（cTnT）的含量，逆轉錄-聚閤酶鏈反應（RT-PCR）、蛋白質免疫印跡試驗（Western Blot）檢測心肌組織HMGB1、單覈細胞趨化蛋白-1（MCP-1）、TNF-α的mRNA及蛋白錶達水平。結果光鏡下可見心肌I/R模型組心肌纖維部分斷裂，心肌細胞大片狀壞死，邊界不清，細胞齣現濃縮、破裂、溶解、甚至消失，間質水腫併伴大量炎性細胞浸潤；針灸預處理組上述錶現較心肌I/R模型組明顯減輕。與假手術組比較，心肌I/R模型組HMGB1、TNF-α、cTnT含量和組織病理學評分均明顯升高〔HMGB1（μg/L）：9.64±1.16比2.15±.031，TNF-α（μg/L）：91±22比19±5， cTnT（μg/L）：1.50±0.35比0.07±0.03，組織病理學評分（分）：2.5±0.3比0.0±0.0，均P＜0.01〕，HMGB1、MCP-1、TNF-αmRNA和蛋白錶達均明顯升高（HMGB1 mRNA：1.42±0.16比0.02±0.00，MCP-1 mRNA：0.46±0.06比0.01±0.00，TNF-αmRNA：0.75±0.04比0.03±0.00；HMGB1蛋白：1.08±0.01比0.02±0.01， MCP-1蛋白：0.92±0.03比0.40±0.01，TNF-α蛋白：1.10±0.02比0.35±0.01，P＜0.05或P＜0.01）；與心肌I/R模型組比較，針灸預處理組HMGB1（6.58±0.73）、TNF-α（63±19）、cTnT（1.15±0.31）含量均明顯降低（均P＜0.01），HMGB1、MCP-1、TNF-αmRNA和蛋白錶達明顯降低（mRNA錶達分彆為0.74±0.12、0.18±0.02、0.10±0.03，蛋白錶達分彆為0.40±0.01、0.36±0.02、0.50±0.02，均P＜0.05），組織病理學評分（1.2±1.0）明顯降低（P＜0.01）。結論針灸預處理可減輕大鼠心肌I/R損傷，其機製可能與減輕HMGB1介導的晚期炎癥反應有關。
목적：탐토침구예처리대대서심기결혈/재관주（I/R）후적보호작용이급대고천이솔족단백B1（HMGB1）표체적영향。방법선택60지Wistar대서，안수궤수자표법분위가수술조、심기I/R모형조、침구예처리조，매조20지。채용결찰관상동맥좌실지좌심이하연약0.5 cm처조단혈류10 min후재관주1 h제비I/R손상모형；가수술조부천선불결찰；침구예처리조우I/R전7 d급여매일1차전침내관혈20 min，련속치료7 d。채용소목소-이홍（HE）염색，광경하관찰심기조직병이학변화，반정량적분법계산3조심기조직병이학평분；채용매련면역흡부시험（ELISA）검측혈장HMGB1、종류배사인자-α（TNF-α）、심기기개단백T（cTnT）적함량，역전록-취합매련반응（RT-PCR）、단백질면역인적시험（Western Blot）검측심기조직HMGB1、단핵세포추화단백-1（MCP-1）、TNF-α적mRNA급단백표체수평。결과광경하가견심기I/R모형조심기섬유부분단렬，심기세포대편상배사，변계불청，세포출현농축、파렬、용해、심지소실，간질수종병반대량염성세포침윤；침구예처리조상술표현교심기I/R모형조명현감경。여가수술조비교，심기I/R모형조HMGB1、TNF-α、cTnT함량화조직병이학평분균명현승고〔HMGB1（μg/L）：9.64±1.16비2.15±.031，TNF-α（μg/L）：91±22비19±5， cTnT（μg/L）：1.50±0.35비0.07±0.03，조직병이학평분（분）：2.5±0.3비0.0±0.0，균P＜0.01〕，HMGB1、MCP-1、TNF-αmRNA화단백표체균명현승고（HMGB1 mRNA：1.42±0.16비0.02±0.00，MCP-1 mRNA：0.46±0.06비0.01±0.00，TNF-αmRNA：0.75±0.04비0.03±0.00；HMGB1단백：1.08±0.01비0.02±0.01， MCP-1단백：0.92±0.03비0.40±0.01，TNF-α단백：1.10±0.02비0.35±0.01，P＜0.05혹P＜0.01）；여심기I/R모형조비교，침구예처리조HMGB1（6.58±0.73）、TNF-α（63±19）、cTnT（1.15±0.31）함량균명현강저（균P＜0.01），HMGB1、MCP-1、TNF-αmRNA화단백표체명현강저（mRNA표체분별위0.74±0.12、0.18±0.02、0.10±0.03，단백표체분별위0.40±0.01、0.36±0.02、0.50±0.02，균P＜0.05），조직병이학평분（1.2±1.0）명현강저（P＜0.01）。결론침구예처리가감경대서심기I/R손상，기궤제가능여감경HMGB1개도적만기염증반응유관。
Objective To investigate the protective effect of electroacupuncture pretreatment on myocardial ischemia/reperfusion (I/R) injury and its influence on high mobility group box 1 (HMGB1) expression in rats. Methods Sixty Wistar rats were randomly divided into sham operation group, myocardial I/R model group and electroacupuncture pretreatment group by random number table (each n = 20). Myocardial I/R injury model was reproduced by ligating the left ventricular branch coronary artery at about 0.5 cm below the atrial appendage lower margin for 10 minutes to occlude the blood flow, then the ligature was relaxed for 1 hour reperfusion; in electroacupuncture pretreatment group, 7 days before I/R, the electroacupuncture at Neiguan acupoint was applied once daily for 20 minutes till the 7th day when I/R was established. Under light microscope, the pathological changes of myocardial specimen stained by hematoxylin-eosine (HE) method were observed. The myocardial histopathological integral was detected by semi quantitative integral method, and the changes of histological scores in three groups were investigated. The levels of plasma HMGB1, tumor necrosis factor-α (TNF-α), cardiac troponin T (cTnT) were detected by enzyme-labeled immunosorbent assay (ELISA). The expressions of HMGB1, monocyte chemotactic protein-1 (MCP-1), TNF-αmRNA and protein in myocardium were detected by reverse transcription-polymerase chain reaction (PT-PCR) and Western Blot. Results Under light microscope, the myocardial tissue in myocardial I/R model group showed partial fracture of myocardial fibers, large patches of myocardial cell necrosis, hazy boundary, cellular condensation, rupture and dissolution or even disappearance, interstitial edema with a lot of inflammatory cell infiltration; the above myocardial tissue injury in electroacupuncture pretreatment group was significantly milder than that in myocardial I/R model group. Compared with sham operation group, in myocardial I/R model group the HMGB1, TNF-α, cTnT contents and histological score were significantly increased [HMGB1 (μg/L):9.64±1.16 vs. 2.15±0.31, TNF-α(μg/L):91±22 vs. 19±5, cTnT (μg/L):1.50±0.35 vs. 0.07±0.03, histological score:2.5±0.3 vs. 0.0±0.0, all P<0.01], HMGB1, MCP-1, TNF-α mRNA and protein expressions were increased obviously (HMGB1 mRNA: 1.42±0.16 vs. 0.02±0.00, MCP-1 mRNA:0.46±0.06 vs. 0.01±0.00, TNF-αmRNA:0.75±0.04 vs. 0.03±0.00;HMGB1 protein:1.08±0.01 vs. 0.20±0.01, MCP-1 protein:0.92±0.03 vs. 0.40±0.01, TNF-αprotein:1.10±0.02 vs. 0.35±0.01, P<0.05 or P<0.01);compared with myocardial I/R model group, in electroacupuncture pretreatment group, HMGB1 (6.58±0.73), TNF-α (63±19), cTnT (1.15±0.31) levels were significantly decreased (all P < 0.01), HMGB1, MCP-1, TNF-αmRNA and protein expressions were markedly reduced (mRNA expression was 0.74±0.12, 0.18±0.02, 0.10±0.03, and protein expression was 0.40±0.01, 0.36±0.02, 0.50±0.02, respectively all P<0.05), and histological score (1.2±1.0) was remarkably lowered (P < 0.01). Conclusion Electroacupuncture pretreatment may reduce the myocardial I/R injury in rats, and the mechanism may be related to the amelioration of inflammatory response mediated by HMGB1 at late stage.
