IL-6及GATA-6甲基化抑制剂在动脉型肺动脉高压大鼠肺组织中的作用及意义  

Role and significance of IL-6 and GATA-6 methylation inhibitors in lung tissue of rats with pulmonary arterial hypertension

作　　者：廖剑雄[1] 罗杰 刘琳[3] 张谦[1] 刘维佳[3] LIAO Jian-xiong;LUO Jie;LIU Lin;ZHANG Qian;LIU Wei-jia(Department of Emergency,Guizhou Provincial People's Hospital,Guiyang 550002,China;School of Medicine,GuiZhou University,Guiyang 550003,China;Department of Respiratory and Critical Care Medicine,Guizhou Provincial People's Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院急诊内科,贵州贵阳550002 [2]贵州大学医学院,贵州贵阳550003 [3]贵州省人民医院呼吸与危重症医学科,贵州贵阳550002

出　　处：《中国病理生理杂志》2022年第3期517-525,共9页Chinese Journal of Pathophysiology

基　　金：贵州省科技计划项目(黔科合基础[2018]1408号)。

摘　　要：目的:通过野百合碱(MCT)诱导的大鼠动脉型肺动脉高压(PAH)模型研究给予白细胞介素6(IL-6)及GATA结合蛋白6(GATA-6)甲基化抑制剂后大鼠肺组织中IL-6、GATA-6及GATA-6甲基化的变化及意义。方法:将40只成年雄性SD大鼠随机分成对照组(按60 mg/kg单次腹腔内注射2∶8的无水乙醇生理盐水混合液)、MCT组(1%MCT按60 mg/kg单次腹腔内注射)、MCT+二甲基亚砜(DMSO)组(等量1%MCT腹腔内注射后每周3次腹部皮下注射1 mg/kg DMSO)、MCT+IL-6抗体(anti-IL-6)组(等量1%MCT腹腔内注射后每天腹部皮下注射40μg/kg anti-IL-6)和MCT+5-氮杂-2′-脱氧胞苷(5-Aza)组(等量1%MCT腹腔内注射后每周3次皮下注射1 mg/kg 5-Aza),每组8只。于造模后第21天做右心室压力监测,计算右心室肥厚指数,肺组织HE染色观察肺病理改变,免疫荧光及Western blot检测肺组织中IL-6和GATA-6的表达,甲基化特异性PCR(MSP)检测肺组织中GATA-6甲基化程度。结果:与对照组比较,MCT组、MCT+anti-IL-6组和MCT+5-Aza组右心室平均压力显著升高(P<0.05);与MCT组比较,MCT+anti-IL-6组和MCT+5-Aza组右心室平均压力显著降低(P<0.05)。与对照组比较,MCT组右心室肥厚指数显著增高(P<0.05);与MCT组比较,MCT+anti-IL-6组和MCT+5-Aza组右心室肥厚指数显著减低(P<0.05)。IL-6免疫荧光及Western blot结果显示,与对照组比较,MCT组和MCT+5-Aza组IL-6表达显著增多(P<0.05);与MCT组比较,MCT+anti-IL-6组和MCT+5-Aza组IL-6表达显著减少(P<0.05)。GATA-6免疫荧光及Western blot结果显示,与对照组比较,MCT组和MCT+5-Aza组GATA-6表达显著减少(P<0.05);与MCT组比较,MCT+anti-IL-6组和MCT+5-Aza组GATA-6表达显著增多(P<0.05)。MSP结果显示,对照组肺组织中GATA-6未发生明显甲基化;MCT组及MCT+DMSO组GATA-6呈高度甲基化;MCT+anti-IL-6组及MCT+5-Aza组GATA-6呈低度甲基化。结论:在PAH大鼠肺组织中给予anti-IL-6后IL-6的表达显著减少,给予5-Aza后GATA-6的表达显著增多,而GATA-6甲基化AIM:To study the changes and significance of interleukin-6(IL-6),GATA-binding protein-6(GATA-6)and GATA-6 methylation in monocrotaline(MCT)-induced rat pulmonary arterial hypertension(PAH)model.METHODS:Forty adult male SD rats were randomly divided into control group,MCT group,MCT+dimethyl sulfoxide(DMSO)group,MCT+IL-6 antibody(anti-IL-6)group and MCT+5-aza-2′-deoxycytidine(5-Aza)group,with 8 rats in each group.The rats in control group received a single intraperitoneal injection of 2∶8 anhydrous ethanol and physiological saline mixture at 60 mg/kg.The rats in MCT group received a single intraperitoneal injection of 1% MCT at 60 mg/kg.The rats in MCT+DMSO group were intraperitoneally injected with the same amount of 1% MCT and subcutaneously injected DMSO at 1 mg/kg into the abdomen 3 times a week.The rats in MCT+anti-IL-6 group were intraperitoneally injected with the same amount of 1% MCT and then intraperitoneally injected with anti-IL-6 at 40μg/kg every day.The rats in MCT+5-Aza group were intraperitoneally injected with the same amount of 1% MCT and then injected subcutaneously with 5-Aza at1 mg/kg 3 times a week.On the 21st day after modeling,the rats in each group were monitored for right ventricular pressure and right ventricular hypertrophy index.Lung tissue HE staining was performed to observe lung pathological changes.The IL-6 and GATA-6 expression in lung tissues was detected by immunofluorescence and Western blot.The GATA-6methylation in lung tissues was detected by methylation-specific PCR(MSP).RESULTS:Compared with control group,the average right ventricular pressure in MCT,MCT+anti-IL-6 and MCT+5-Aza groups was significantly increased(P<0.05).Compared with MCT group,the average pressure of right ventricle in MCT+anti-IL-6 and MCT+5-Aza groups was significantly decreased(P<0.05).Compared with control group,the right ventricular hypertrophy index in MCT group was significantly increased(P<0.05).Compared with MCT group,the right ventricular hypertrophy index in MCT+antiIL-6 and MCT+5-Aza groups

关 键 词：动脉型肺动脉高压 白细胞介素6 GATA结合蛋白6 5-氮杂-2′-脱氧胞苷 

分 类 号：R543.1[医药卫生—心血管疾病] R363.2[医药卫生—内科学]