CAJ | 학술논문

目的：观察绿原酸对SD大鼠心肌缺血－再灌注损伤的防护作用并分析其作用机制。方法选取清洁级健康雄性SD大鼠16只（200～220 g）。随机分为缺血－再灌注组（I/R组）和绿原酸干预组（CGA＋I/R组），每组8只。适应性饲养1周后，CGA＋I/R组给予灌胃绿原酸50 mg/kg，I/R组给予同样方法灌等量生理盐水，连续饲养2周后建立在体心脏缺血－再灌注损伤模型，各组均缺血30 min，再灌注24 h后收集标本，心脏标本留取进行氯化三苯基四氮唑法（TTC）染色观察心肌梗死面积，酶联免疫吸附试验( ELSIA )法检测大鼠血清中乳酸脱氢酶（LDH）含量。同时，应用H9C2细胞建立缺氧复氧损伤模型（缺氧8 h，复氧24 h），随机分为3组，即对照组、缺氧复氧组（A/R组）和绿原酸干预组（CGA＋A/R组）。复氧结束后，收集上清液及细胞，上清液利用ELISA法检测炎症因子白细胞介素－6（IL－6）和肿瘤坏死因子－α（TNF－α），应用qRT－PCR检测收集细胞的IL－6及TNF－αmRNA含量，96孔板中细胞应用CCK－8检测细胞活力。结果与I/R组比较，CGA＋I/R组能显著减少心肌梗死面积[（23.97±2.8)％比(16.2±2.1)％，P＜0.05）]，抑制心肌酶释放[（5038±257.6) U/L比(4005±206.8) U/L，P＜0.05）]；与 A/R组比较，CGA＋A/R组能显著减少细胞的损伤[（51.6±7.8)％比(69.7±6.6)％，P＜0.05）]，抑制炎症因子的释放（P＜0.05）。结论绿原酸可通过抑制炎症因子的释放从而发挥对心肌缺血－再灌注损伤的防护作用。
목적：관찰록원산대SD대서심기결혈－재관주손상적방호작용병분석기작용궤제。방법선취청길급건강웅성SD대서16지（200～220 g）。수궤분위결혈－재관주조（I/R조）화록원산간예조（CGA＋I/R조），매조8지。괄응성사양1주후，CGA＋I/R조급여관위록원산50 mg/kg，I/R조급여동양방법관등량생리염수，련속사양2주후건립재체심장결혈－재관주손상모형，각조균결혈30 min，재관주24 h후수집표본，심장표본류취진행록화삼분기사담서법（TTC）염색관찰심기경사면적，매련면역흡부시험( ELSIA )법검측대서혈청중유산탈경매（LDH）함량。동시，응용H9C2세포건립결양복양손상모형（결양8 h，복양24 h），수궤분위3조，즉대조조、결양복양조（A/R조）화록원산간예조（CGA＋A/R조）。복양결속후，수집상청액급세포，상청액이용ELISA법검측염증인자백세포개소－6（IL－6）화종류배사인자－α（TNF－α），응용qRT－PCR검측수집세포적IL－6급TNF－αmRNA함량，96공판중세포응용CCK－8검측세포활력。결과여I/R조비교，CGA＋I/R조능현저감소심기경사면적[（23.97±2.8)％비(16.2±2.1)％，P＜0.05）]，억제심기매석방[（5038±257.6) U/L비(4005±206.8) U/L，P＜0.05）]；여 A/R조비교，CGA＋A/R조능현저감소세포적손상[（51.6±7.8)％비(69.7±6.6)％，P＜0.05）]，억제염증인자적석방（P＜0.05）。결론록원산가통과억제염증인자적석방종이발휘대심기결혈－재관주손상적방호작용。
Objective To investigate the cardioProtective effects of chlorogenic acid against myocardial ischemia/rePerfusion injury and to analyze its mechanism. Methods 16 healthy SD rats of clean grade(200~220 g)were selected and randomly devided into two grouPs:ischemia rePerfusion grouP (I/R grouP) and chlorogenic acid (CGA+I/R) grouP,8 rats in each grouP. After one week's adaPtive feeding,rats in the CGA+I/R grouP were gavaged with 50 mg/kg CGA,while rats in the I/R grouP were gavaged with equivalent normal saline for 2 weeks. In vivo cardiac ischemia/rePerfusion injury model were then built in each rat. All heart samPles were col-lected after 30 mins' ischemia and 24 hours' rePerfusion,and dyed with triPhenyltetrazolium chloride (TTC) to observe the infarction area of myocardium. LDH content was also measured in rats serum by ELISA method. H9C2 cells were used to build anoxia/reoxygena-tion(A/R)injury model(8 hours' anoxia and 24 hours' reoxygenation). Cells were randomly devided into 3 grouPs:the control grouP, A/R grouP and CGA+A/R grouP. SuPerate and cells were resPectively collected after reoxygenation. SuPernate was used to detect in-flammatory factors like IL-6 and TNF-α by ELISA method,cells were used to quantify mRNA of IL-6 and TNF-α by qRT-PCR. 96-well Plates cells were used to test cell viability by using CCK-8 method. Results ComPared with the I/R grouP,the CGA+I/R grouP had significantly less infarction area [(23. 97 ± 2. 8 )% vs. ( 16. 2 ± 2. 1 )%,P < 0. 05),less myocardial enzyme release [(5 038 ± 257. 6 ) U/L vs. ( 4 005 ± 206. 8 ) U/L,P < 0. 05)];when comPared with the A/R grouP,the CGA+A/R grouP could remarkably reduce cell injury [(51. 6 ± 7. 8 )% vs. ( 69. 7 ± 6. 6 )%,P < 0. 05)] ,and inhibit the release of inflammatory factors. Conclusion CGA Plays an Protective effects on myocardial I/R injury via inhibiting the release of inflammatory factors.