CAJ | 학술논문

目的探讨烟碱对心肌缺血/再灌注(I/R)损伤大鼠炎症细胞因子的影响.方法 50只健康雄性SD大鼠按随机数字表法分为假手术组、I/R组、烟碱高剂量(400μg/kg)组、烟碱低剂量(40μg/kg)组及α-银环蛇毒素(α-BGT,1μg/kg)组5组,每组10只.采用结扎心脏左冠状动脉前降支30 min、再灌注90 min制作大鼠心肌I/R损伤模型;假手术组仅穿线不结扎.制模前30 min各药物组颈静脉注射相应剂量药物干预,假手术组和I/R组给予等量生理盐水.于再灌注末取右颈动脉血,测定肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、IL-10浓度和肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)活性;然后处死动物,取缺血区心肌组织测定髓过氧化物酶(MPO)活性;采用免疫组化和逆转录-聚合酶链反应检测心肌组织细胞间黏附分子-1(ICAM-1)蛋白及mRNA表达,并观察心肌超微结构.结果与假手术组比较,I/R组血浆TNF-α、IL-8、IL-10、CK-MB、cTnI、心肌MPO活性及ICAM-1蛋白和mRNA表达均显著升高[TNF-α(ng/L):158.7±32.7比31.5±5.8,IL-8(ng/L):0.71±0.06比0.30±0.04,IL-10(ng/L):69.0±7.8比41.4±4.3,CK-MB(U/L):2 540±169比1 120±102,cTnI(μg/L):26.2±4.6比0.9±0.2,MPO(U/g):4.2±0.6比1.6±0.4,ICAM-1蛋白:0.210±0.025比0.100±0.018,ICAM-1 mRNA:1.82±0.23比1.18±0.20,P＜0.05或P＜0.01],病理学显示心肌组织损伤较重.与I/R组比较,烟碱高剂量组血浆TNF-α、IL-8降低[TNF-α(67.3±9.8)ng/L,IL-8(0.47±0.04)ng/L],IL-10升高[(147.5±12.5)ng/L],CK-MB、cTnI及心肌MPO活性、ICAM-1蛋白和mRNA均降低[CK-MB(1 282±145)U/L,cTnI(4.7±1.4)μg/L,MPO(2.5±0.4)U/g,ICAM-1蛋白0.140±0.026,ICAM-1 mRNA 1.31±0.25,P＜0.05或P＜0.01],心肌组织损伤减轻;而烟碱低剂量组和α-BGT组上述指标与I/R组比较差异无统计学意义.结论烟碱可阻断内皮细胞表达黏附分子,阻断中性粒细胞黏附、游出,改善抗炎/促炎反应平衡,从而拮抗大鼠心肌I/R损伤时的过度炎症反应. 目的探討煙堿對心肌缺血/再灌註(I/R)損傷大鼠炎癥細胞因子的影響.方法 50隻健康雄性SD大鼠按隨機數字錶法分為假手術組、I/R組、煙堿高劑量(400μg/kg)組、煙堿低劑量(40μg/kg)組及α-銀環蛇毒素(α-BGT,1μg/kg)組5組,每組10隻.採用結扎心髒左冠狀動脈前降支30 min、再灌註90 min製作大鼠心肌I/R損傷模型;假手術組僅穿線不結扎.製模前30 min各藥物組頸靜脈註射相應劑量藥物榦預,假手術組和I/R組給予等量生理鹽水.于再灌註末取右頸動脈血,測定腫瘤壞死因子-α(TNF-α)、白細胞介素-8(IL-8)、IL-10濃度和肌痠激酶同工酶(CK-MB)、心肌肌鈣蛋白I(cTnI)活性;然後處死動物,取缺血區心肌組織測定髓過氧化物酶(MPO)活性;採用免疫組化和逆轉錄-聚閤酶鏈反應檢測心肌組織細胞間黏附分子-1(ICAM-1)蛋白及mRNA錶達,併觀察心肌超微結構.結果與假手術組比較,I/R組血漿TNF-α、IL-8、IL-10、CK-MB、cTnI、心肌MPO活性及ICAM-1蛋白和mRNA錶達均顯著升高[TNF-α(ng/L):158.7±32.7比31.5±5.8,IL-8(ng/L):0.71±0.06比0.30±0.04,IL-10(ng/L):69.0±7.8比41.4±4.3,CK-MB(U/L):2 540±169比1 120±102,cTnI(μg/L):26.2±4.6比0.9±0.2,MPO(U/g):4.2±0.6比1.6±0.4,ICAM-1蛋白:0.210±0.025比0.100±0.018,ICAM-1 mRNA:1.82±0.23比1.18±0.20,P＜0.05或P＜0.01],病理學顯示心肌組織損傷較重.與I/R組比較,煙堿高劑量組血漿TNF-α、IL-8降低[TNF-α(67.3±9.8)ng/L,IL-8(0.47±0.04)ng/L],IL-10升高[(147.5±12.5)ng/L],CK-MB、cTnI及心肌MPO活性、ICAM-1蛋白和mRNA均降低[CK-MB(1 282±145)U/L,cTnI(4.7±1.4)μg/L,MPO(2.5±0.4)U/g,ICAM-1蛋白0.140±0.026,ICAM-1 mRNA 1.31±0.25,P＜0.05或P＜0.01],心肌組織損傷減輕;而煙堿低劑量組和α-BGT組上述指標與I/R組比較差異無統計學意義.結論煙堿可阻斷內皮細胞錶達黏附分子,阻斷中性粒細胞黏附、遊齣,改善抗炎/促炎反應平衡,從而拮抗大鼠心肌I/R損傷時的過度炎癥反應.
목적 탐토연감대심기결혈/재관주(I/R)손상대서염증세포인자적영향.방법 50지건강웅성SD대서안수궤수자표법분위가수술조、I/R조、연감고제량(400μg/kg)조、연감저제량(40μg/kg)조급α-은배사독소(α-BGT,1μg/kg)조5조,매조10지.채용결찰심장좌관상동맥전강지30 min、재관주90 min제작대서심기I/R손상모형;가수술조부천선불결찰.제모전30 min각약물조경정맥주사상응제량약물간예,가수술조화I/R조급여등량생리염수.우재관주말취우경동맥혈,측정종류배사인자-α(TNF-α)、백세포개소-8(IL-8)、IL-10농도화기산격매동공매(CK-MB)、심기기개단백I(cTnI)활성;연후처사동물,취결혈구심기조직측정수과양화물매(MPO)활성;채용면역조화화역전록-취합매련반응검측심기조직세포간점부분자-1(ICAM-1)단백급mRNA표체,병관찰심기초미결구.결과 여가수술조비교,I/R조혈장TNF-α、IL-8、IL-10、CK-MB、cTnI、심기MPO활성급ICAM-1단백화mRNA표체균현저승고[TNF-α(ng/L):158.7±32.7비31.5±5.8,IL-8(ng/L):0.71±0.06비0.30±0.04,IL-10(ng/L):69.0±7.8비41.4±4.3,CK-MB(U/L):2 540±169비1 120±102,cTnI(μg/L):26.2±4.6비0.9±0.2,MPO(U/g):4.2±0.6비1.6±0.4,ICAM-1단백:0.210±0.025비0.100±0.018,ICAM-1 mRNA:1.82±0.23비1.18±0.20,P＜0.05혹P＜0.01],병이학현시심기조직손상교중.여I/R조비교,연감고제량조혈장TNF-α、IL-8강저[TNF-α(67.3±9.8)ng/L,IL-8(0.47±0.04)ng/L],IL-10승고[(147.5±12.5)ng/L],CK-MB、cTnI급심기MPO활성、ICAM-1단백화mRNA균강저[CK-MB(1 282±145)U/L,cTnI(4.7±1.4)μg/L,MPO(2.5±0.4)U/g,ICAM-1단백0.140±0.026,ICAM-1 mRNA 1.31±0.25,P＜0.05혹P＜0.01],심기조직손상감경;이연감저제량조화α-BGT조상술지표여I/R조비교차이무통계학의의.결론 연감가조단내피세포표체점부분자,조단중성립세포점부、유출,개선항염/촉염반응평형,종이길항대서심기I/R손상시적과도염증반응.
Objective To investigate the effect of nicotine on inflammatory cytokines in myocardial ischemia/reperfusion (I/R) injury in rat. Methods Fifty male Sprague-Dawley (SD) rats were divided into five groups by random numbers table (each n= 10): sham operation group (S group), I/R group, nicotine 400 μg/kg group (H group), nicotine 40 μg/kg group (L group) and α-bungarotoxin (α-BGT, 1 μg/kg)group. The anterior descending branch of left coronary artery was occluded for 30 minutes followed by 90 minutes reperfusion to reproduce myocardial I/R injury rat model, while in S group the anterior descending branch of left coronary artery was only exposed without occlusion procedure. Thirty minutes before myocardial ischemia, drugs in corresponding doses were given intravenously via jugular vein. At the end of 90 minutes of reperfusion, blood samples were collected from carotid artery to determine the levels of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), IL-10, MB isoenzyme of creatine kinase (CK-MB),and cardiac troponin I (cTnI), then the animals were sacrificed and the hearts were harvested for pathological study and determination of myeloperoxidase (MPO) activity. Immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) were used to assess intercellular adhesion molecule-1 (ICAM-1) protein and mRNA expression in heart tissue. Results Compared with the S group, the concentrations of TNF-α, IL-8, IL-10, CK-MB, cTnI, MPO activity, ICAM-1 protein and mRNA expression were significantly increased in I/R group [TNF-α (ng/L): 158. 7± 32. 7 vs. 31.5±5.8, IL-8(ng/L): 0.71±0.06 vs. 0. 30±0. 04, IL-10 (ng/L): 69.0±7.8 vs. 41.4±4.3, CK-MB (U/L): 2540±169 vs. 1 120±120, cTnI (μg/L): 26.2±4.6 vs. 0.9±0.2, MPO (U/g): 4.2±0.6 vs. 1.6±0.4,ICAM-1 protein: 0. 210±0. 025 vs. 0. 100±0. 018, ICAM-1 mRNA: 1.82±0.23 vs. 1.18±0. 20, P＜0. 05or P＜0. 01]. Injury to myocardial ultrastructure was worse in I/R group. Compared with the I/R group,the plasma levels of TNF-α and IL-8 were lower [TNF-α (67.3±9.8) ng/L, IL-8 (0. 47±0. 04) ng/L],IL-10 was higher [(147.5± 12.5) ng/L], CK-MB, cTnI, MPO, ICAM-1 protein and mRNA were lower obviously in H group [CK-MB (1 282±145) U/L, cTnI (4. 7±1.4) μg/L, MPO (2. 5±0. 4) U/g, ICAM-1 protein 0. 140 ± 0. 026, ICAM-1 mRNA 1.31 ± 0.25, P ＜ 0. 05 or P ＜ 0. 01]. Injury to the myocardial ultrastructure was less marked in H group. The indexes of those in L group and α-BGT group compared with I/R group were not statistically significantly different. Conclusion Nicotine can block endothelial expression of adhesion molecules and neutrophil adhesion and infiltration to promote a balance of anti-inflammatory and pro-inflammatory response, thus prevents excessive inflammatory response to myocardial I/R injury in rat.