CAJ | 학술논문

目的：观察 N-乙酰半胱氨酸（NAC）预处理对心脏移植缺血再灌注损伤（IRI）时基质金属蛋白酶（MMP）的影响，并探讨其可能的机制。方法健康雄性 Lewis 大鼠60只，随机分为3组。对照组：摘取供体心脏前30 min 经供体大鼠腹腔静脉注射生理盐水0.5 ml；供体预处理组：摘取供心前30 min 经供体大鼠腹腔静脉注射 NAC 300 mg／kg；受体预处理组：受体于移植前30 min 经受体大鼠腹腔静脉注射 NAC 300 mg／kg。供心冷藏于4℃组氨酸-色氨酸-酮戊二酸盐液（HTK）18 h后建立大鼠腹腔异体异位心脏移植模型。移植后24 h 光镜下观察心肌胶原含量的变化；免疫组织化学法检测心肌组织中 MMP2／9蛋白表达；采用实时 PCR 检测 MMP2和基质金属蛋白酶抑制剂2（TIMP2）mRNA 表达。结果NAC 供体／受体预处理组心肌胶原含量明显低于对照组（4.49±0.62／5.88±0.96 vs.10.43±0.93，P ＜0.01）；免疫组织化学法半定量评分显示 NAC 供体／受体预处理组MMP2／9蛋白含量明显低于对照组（1.60±0.22／2.13±0.23，1.63±0.18／2.78±0.16 vs.3.00±0.15／3.78±0.15，P ＜0.01），受体预处理组心肌组织中 MMP2 mRNA 明显低于对照组（1.45±0.72 vs.2.77±1.62，P ＜0.05），而供体预处理组 TIMP2 mRNA 明显高于对照组（2.59±2.12 vs.0.63±0.50，P ＜0.05）。结论在大鼠心脏移植中 NAC 对供心 IRI 具有保护作用，可能是通过 NAC 直接抑制活性氧所致的 MMP2／9升高，激活 TIMP2，减轻了心肌 IRI。
목적：관찰 N-을선반광안산（NAC）예처리대심장이식결혈재관주손상（IRI）시기질금속단백매（MMP）적영향，병탐토기가능적궤제。방법건강웅성 Lewis 대서60지，수궤분위3조。대조조：적취공체심장전30 min 경공체대서복강정맥주사생리염수0.5 ml；공체예처리조：적취공심전30 min 경공체대서복강정맥주사 NAC 300 mg／kg；수체예처리조：수체우이식전30 min 경수체대서복강정맥주사 NAC 300 mg／kg。공심랭장우4℃조안산-색안산-동무이산염액（HTK）18 h후건립대서복강이체이위심장이식모형。이식후24 h 광경하관찰심기효원함량적변화；면역조직화학법검측심기조직중 MMP2／9단백표체；채용실시 PCR 검측 MMP2화기질금속단백매억제제2（TIMP2）mRNA 표체。결과NAC 공체／수체예처리조심기효원함량명현저우대조조（4.49±0.62／5.88±0.96 vs.10.43±0.93，P ＜0.01）；면역조직화학법반정량평분현시 NAC 공체／수체예처리조MMP2／9단백함량명현저우대조조（1.60±0.22／2.13±0.23，1.63±0.18／2.78±0.16 vs.3.00±0.15／3.78±0.15，P ＜0.01），수체예처리조심기조직중 MMP2 mRNA 명현저우대조조（1.45±0.72 vs.2.77±1.62，P ＜0.05），이공체예처리조 TIMP2 mRNA 명현고우대조조（2.59±2.12 vs.0.63±0.50，P ＜0.05）。결론재대서심장이식중 NAC 대공심 IRI 구유보호작용，가능시통과 NAC 직접억제활성양소치적 MMP2／9승고，격활 TIMP2，감경료심기 IRI。
Objective To explore the protective effect of N-acetylcysteine (NAC)on modulating ma-trix metalloproteinase (MMP)in myocardial ischemia reperfusion injury (IRI)during heart transplantation in rat models.Methods Sixty healthy male Lewis rats were randomly divided into three groups.Control group:0.5 ml Saline was infused via inferior vena cava 30 min before donor harvesting;Donor pretreatment group:NAC (300 mg/kg BW)was infused via inferior vena cava 30 min before donor harvesting;Recipient pretreat-ment group:NAC (300 mg/kg BW)was infused via inferior vena cava 30min before recipient implantation.Grafts were kept in cold storage in conventional histidine tryptophan ketoglutarate (HTK)solution at 4℃ for 1 8 hrs prior to heterotopic transplantation.Heart transplantation was performed in each group.Tissue samples were taken 24 hrs after reperfusion for measurement of tissue collagen contents,MMP2 /9 protein expression,and MMP2 /TIMP2 mRNA expression.Tissue collagen contents were detected by sirus red.MMP2 /9 protein ex-pression was detected by immunohistochemistry and MMP2 /TIMP2 mRNA expression was measured by real-time PCR.Results Tissue collagen contents in donor/recipient pretreatment group significantly decreased compared with that in control group (4.49 ±0.62 /5.88 ±0.96 vs.1 0.43 ±0.93,P <0.01 ).MMP2 /9 pro-tein expression in donor/recipient pretreatment group was significantly lower compared with that in control group (1 .60 ±0.22 /2.1 3 ±0.23,1 .63 ±0.1 8 /2.78 ±0.1 6 vs.3.00 ±0.1 5 /3.78 ±0.1 5,P <0.01 ),as illustra-ted in Table 3.Real-time PCR revealed that the level of MMP2 mRNA in recipient pretreatment group was sig-nificantly lower compared with that in control group (1 .45 ±0.72 vs.2.77 ±1 .62,P <0.05),whereas the expression of TIMP2 mRNA in donor pretreatment group was significantly higher than that in control group (2.59 ±2.1 2 vs.0.63 ±0.50,P <0.05).Conclusion NAC can protect graft cardiac function after heart transplantation,probably by directly suppressing reactive oxygen species-induced MMP2 /9 evelation and up-regulating TIMP2 activity.