中国临床康复
中國臨床康複
중국림상강복
CHINESE JOURNAL OF CLINICAL REHABILITATION
2005年
47期
172-173
,共2页
孟庆涛%夏中元%刘先义%陈向东%熊桂先
孟慶濤%夏中元%劉先義%陳嚮東%熊桂先
맹경도%하중원%류선의%진향동%웅계선
参附汤/治疗应用%再灌注损伤%小肠%肿瘤坏死因子
參附湯/治療應用%再灌註損傷%小腸%腫瘤壞死因子
삼부탕/치료응용%재관주손상%소장%종류배사인자
背景:肠上皮细胞异常凋亡是缺血再灌注期间肠黏膜损伤的主要原因,参附注射液对小肠黏膜损伤有良好的防治作用.目的:建立大鼠肠缺血再灌注模型,观察缺血再灌注时给予参附注射液对肠上皮细胞凋亡数目、凋亡相关基因半胱氨酸天冬氨酸蛋白酶-3、Bcl-2的表达及肿瘤坏死因子水平的影响.设计:随机对照实验.单位:武汉大学人民医院麻醉科.材料:实验于2002-12/2003-06在武汉大学人民医院麻醉科实验室完成.选取健康清洁级SD大鼠24只,随机分为空白对照组、肠缺血再灌注组、参附注射液组,8只/组.方法:各组大鼠给予氨基甲酸乙酯1 mg/kg腹腔注射麻醉,肠缺血再灌注组和参附注射液组用血管钳夹闭肠系膜上动脉1 h然后再灌注2 h,空白对照组不用血管钳夹闭肠系膜上动脉.参附注射液组于阻断前30 min静注参附注射液0.02 mL/g,空白对照组和肠缺血再灌注组静脉输注等量生理盐水.制备切片进行半胱氨酸天冬氨酸蛋白酶-3、Bcl-2蛋白的表达及细胞凋亡检测.主要观察指标:①各组大鼠细胞凋亡指数的比较.②各组大鼠肠上皮细胞半胱氨酸天冬氨酸蛋白酶-3及Bcl-2基因的表达.③各组肠粘膜匀浆中肿瘤坏死因子含量的比较.结果:实验选用24只大鼠,全部进入结果分析.①各组大鼠细胞凋亡指数的比较:参附注射液组的凋亡指数明显低于肠缺血再灌注组,但比空白对照组高[(7.75±1.89)%,(28.25±8.50)%,(4.25±2.63)%,P均＜0.01].②各组大鼠肠上皮细胞半胱氨酸天冬氨酸蛋白酶-3及Bcl-2基因的表达:参附注射液组组半胱氨酸天冬氨酸蛋白酶-3的表达低于肠缺血再灌注组,但高于空白对照组[(0.211 6±0.087 5),(0.354 7±0.077 8),(0.194 1±0.057 4),P＜0.01,P＞0.05];Bcl-2的表达在肠缺血再灌注组比空白对照组显著增高(P＜0.05),参附注射液组的表达比肠缺血再灌注组明显降低(P＜0.01).③各组肠黏膜匀浆中肿瘤坏死因子含量的比较:砀缺血再灌注组肿瘤坏死因子的表达显著高于空白对照组和参附注射液组[(189.7±56.3),(38.6±10.4),(47.5±18.7)mg/L,P均＜0.01],参附注射液组和空白对照组基本相似(P＞0.05).结论:参附注射液通过抑制肿瘤坏死因子的含量、降低半胱氨酸天冬氨酸蛋白酶-3的表达、上调Bcl-2基因而抑制缺血再灌注期间肠黏膜上皮细胞的凋亡,从而减轻肠黏膜缺血再灌注损伤.
揹景:腸上皮細胞異常凋亡是缺血再灌註期間腸黏膜損傷的主要原因,參附註射液對小腸黏膜損傷有良好的防治作用.目的:建立大鼠腸缺血再灌註模型,觀察缺血再灌註時給予參附註射液對腸上皮細胞凋亡數目、凋亡相關基因半胱氨痠天鼕氨痠蛋白酶-3、Bcl-2的錶達及腫瘤壞死因子水平的影響.設計:隨機對照實驗.單位:武漢大學人民醫院痳醉科.材料:實驗于2002-12/2003-06在武漢大學人民醫院痳醉科實驗室完成.選取健康清潔級SD大鼠24隻,隨機分為空白對照組、腸缺血再灌註組、參附註射液組,8隻/組.方法:各組大鼠給予氨基甲痠乙酯1 mg/kg腹腔註射痳醉,腸缺血再灌註組和參附註射液組用血管鉗夾閉腸繫膜上動脈1 h然後再灌註2 h,空白對照組不用血管鉗夾閉腸繫膜上動脈.參附註射液組于阻斷前30 min靜註參附註射液0.02 mL/g,空白對照組和腸缺血再灌註組靜脈輸註等量生理鹽水.製備切片進行半胱氨痠天鼕氨痠蛋白酶-3、Bcl-2蛋白的錶達及細胞凋亡檢測.主要觀察指標:①各組大鼠細胞凋亡指數的比較.②各組大鼠腸上皮細胞半胱氨痠天鼕氨痠蛋白酶-3及Bcl-2基因的錶達.③各組腸粘膜勻漿中腫瘤壞死因子含量的比較.結果:實驗選用24隻大鼠,全部進入結果分析.①各組大鼠細胞凋亡指數的比較:參附註射液組的凋亡指數明顯低于腸缺血再灌註組,但比空白對照組高[(7.75±1.89)%,(28.25±8.50)%,(4.25±2.63)%,P均＜0.01].②各組大鼠腸上皮細胞半胱氨痠天鼕氨痠蛋白酶-3及Bcl-2基因的錶達:參附註射液組組半胱氨痠天鼕氨痠蛋白酶-3的錶達低于腸缺血再灌註組,但高于空白對照組[(0.211 6±0.087 5),(0.354 7±0.077 8),(0.194 1±0.057 4),P＜0.01,P＞0.05];Bcl-2的錶達在腸缺血再灌註組比空白對照組顯著增高(P＜0.05),參附註射液組的錶達比腸缺血再灌註組明顯降低(P＜0.01).③各組腸黏膜勻漿中腫瘤壞死因子含量的比較:碭缺血再灌註組腫瘤壞死因子的錶達顯著高于空白對照組和參附註射液組[(189.7±56.3),(38.6±10.4),(47.5±18.7)mg/L,P均＜0.01],參附註射液組和空白對照組基本相似(P＞0.05).結論:參附註射液通過抑製腫瘤壞死因子的含量、降低半胱氨痠天鼕氨痠蛋白酶-3的錶達、上調Bcl-2基因而抑製缺血再灌註期間腸黏膜上皮細胞的凋亡,從而減輕腸黏膜缺血再灌註損傷.
배경:장상피세포이상조망시결혈재관주기간장점막손상적주요원인,삼부주사액대소장점막손상유량호적방치작용.목적:건립대서장결혈재관주모형,관찰결혈재관주시급여삼부주사액대장상피세포조망수목、조망상관기인반광안산천동안산단백매-3、Bcl-2적표체급종류배사인자수평적영향.설계:수궤대조실험.단위:무한대학인민의원마취과.재료:실험우2002-12/2003-06재무한대학인민의원마취과실험실완성.선취건강청길급SD대서24지,수궤분위공백대조조、장결혈재관주조、삼부주사액조,8지/조.방법:각조대서급여안기갑산을지1 mg/kg복강주사마취,장결혈재관주조화삼부주사액조용혈관겸협폐장계막상동맥1 h연후재관주2 h,공백대조조불용혈관겸협폐장계막상동맥.삼부주사액조우조단전30 min정주삼부주사액0.02 mL/g,공백대조조화장결혈재관주조정맥수주등량생리염수.제비절편진행반광안산천동안산단백매-3、Bcl-2단백적표체급세포조망검측.주요관찰지표:①각조대서세포조망지수적비교.②각조대서장상피세포반광안산천동안산단백매-3급Bcl-2기인적표체.③각조장점막균장중종류배사인자함량적비교.결과:실험선용24지대서,전부진입결과분석.①각조대서세포조망지수적비교:삼부주사액조적조망지수명현저우장결혈재관주조,단비공백대조조고[(7.75±1.89)%,(28.25±8.50)%,(4.25±2.63)%,P균＜0.01].②각조대서장상피세포반광안산천동안산단백매-3급Bcl-2기인적표체:삼부주사액조조반광안산천동안산단백매-3적표체저우장결혈재관주조,단고우공백대조조[(0.211 6±0.087 5),(0.354 7±0.077 8),(0.194 1±0.057 4),P＜0.01,P＞0.05];Bcl-2적표체재장결혈재관주조비공백대조조현저증고(P＜0.05),삼부주사액조적표체비장결혈재관주조명현강저(P＜0.01).③각조장점막균장중종류배사인자함량적비교:탕결혈재관주조종류배사인자적표체현저고우공백대조조화삼부주사액조[(189.7±56.3),(38.6±10.4),(47.5±18.7)mg/L,P균＜0.01],삼부주사액조화공백대조조기본상사(P＞0.05).결론:삼부주사액통과억제종류배사인자적함량、강저반광안산천동안산단백매-3적표체、상조Bcl-2기인이억제결혈재관주기간장점막상피세포적조망,종이감경장점막결혈재관주손상.
BACKGROUND: The abnormal apoptosis of intestinal epithelial cells is the main cause of intestinal mucous membrane injury during ischemia reperfusion. Shenfu injection has good therapeutic effect on intestinal mucous membrane injury. OBJECTIVE: To investigate the influence of shenfu injection medication on intestinal apoptotic epithelial number, apoptosis-related caspase-3 and Bcl-2 gene expression, as well as the level of tumor necrosis factor (TNF)in intestinal ischemia reperfusion rat model.DESIGN: Randomized controlled experiment.SETTING:Department of Anesthesia, Renmin Hospital of Wuhan University.MATERIALS:This experiment was carried out in the laboratory of the Department of Anesthesia, Renmin Hospital of Wuhan University, between December 2002 and June 2003. Totally 24 healthy SD rats of clean grade were randomized into blank control group, intestinal ischemia reperfusion (IR) group and shenfu injection group with 8 rats in each group.METHODS: Rats in each group were anesthetized with intraperitoneal injection of ethyl carbamate at dosage of 1mg/kg; then in intestinal IR group and shenfu injection group rats' superior mesenteric artery was occluded with vascular clamp for 1 hour before 2-hour reperfusion,which was not conducted in blank control group. Rats in shenfu injection group were intravenously injected with shenfu injection at 30 minutes before occlusion at dose of 0.02 mL/g, which was replaced by the same volume of physical saline in blank control group and intestinal IR group. The expression of caspase-3 and Bcl-2 protein and cell apoptosis wee detected.MAIN OUTCOME MEASURES: ① Comparison of apoptotic index between groups. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium. ③ Comparison of TNF content in intestinal mucous membrane homogenate between groups.RESULTS: Totally 24 rats were included in this experiment and all ehtered the result analysis. ①Comparison of apoptotic index between groups:The apoptotic index was obviously lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(7.75-±1.89)%,(28.25±8.50)%, (4.25-±2.63)%, P ＜ 0.01]. ② The expression of caspase-3 and Bcl-2 gene in rat intestinal epithelium: caspase-3 expression was lower in shenfu injection group than in intestinal IR group, but higher than in blank control group [(0.211 6±0.087 5), (0.354 7±0.077 8), (0.194 1±0.057 4) A,P ＜ 0.01, P ＞ 0.05]; Bcl-2 expression was remarkably higher in intestinal IR group than in blank control group (P ＜ 0.05), but obviously reduced in shenfu injection group compared to intestinal IR group (P ＜ 0.01). ③ TNF content of intestinal mucous membranehomogenate in each group: TNF content was remarkably higher in intestinal IR group than in blank control group and shenfu injection group [(189.7±56.3), (38.6±10.4), (47.5±l8.7)mg/L,P ＜ 0.01], and basically the same in shenfu inj~tion group and blank control group (P ＞ 0.05).CONCLUSION: Shenfu injection can suppress intestinal epithelial apoptosis by reducing TNF content and caspase-3 e.pression as well as upregulating the expression of Bcl-2 gene during ischemia reperfusion, thereby attenuating ischemia reperfusion injury of intestinal mucous membrane.
