CAJ | 학술논문

目的探讨缬沙坦与U0126对血管紧张素Ⅱ(angiotensin Ⅱ,Ang Ⅱ)诱导的大鼠心房纤维化和缝隙连接蛋白40( connexin 40,Cx40)重构的影响.方法将32只雄性SD大鼠随机分为空白对照组(A组)、盐酸异丙基肾上腺素( isopreterenol,ISO)+二甲基亚砜(DMSO)组(B组)、ISO+U0126组(C组,U0126溶于DMSO中)、ISO+缬沙坦+DMSO组(D组).给药28 d后处死大鼠取心肌组织,放射免疫法测Ang Ⅱ含量;HE和Masson染色法观察纤维化程度即胶原容积分数(CVF);免疫组化法测定磷酸化丝裂原活化蛋白激酶激酶1/2(P-MEK1/2)、磷酸化细胞外信号调节激酶1/2( P-ERK1/2)以及Cx40的表达.结果 B、C、D组中Ang Ⅱ含量较A组明显升高[分别为(368.243±6.283)ng/L、(357.175±5.944) ng/L、(359.908±2.496) ng/L比(250.380±4.261) ng/L,P＜0.01];A组CVF(9.025 ±0.456)％,显示无心房纤维化;C组和D组较B组心房纤维化程度明显减弱[CVF分别为(10.260±0.525)％、(10.238 ±0.524)％比(78.710±1.587)％,P＜0.01],C组和D组之间差异无统计学意义(P＞0.05);B组较A组中P-MEK1/2(0.311±0.007比0.203±0.009,P＜0.01)和P-ERK1/2含量明显增加(0.259±0.003比0.173±0.006,P＜0.01),而C组和D组中含量较B组明显减少(P-MEK1/2分别为0.212±0.004、0.213±0.005比0.311±0.007,P＜0.01;P-ERK1/2分别为0.178±0.004、0.175 ±0.007比0.259±0.003,P＜0.01),C组和D组之间差异无统计学意义(P＞0.05);B组较A组Cx40含量明显减少(0.199±0.007比0.241±0.004,P＜0.01)且分布紊乱,C组和D组中含量较B组减少程度明显减轻(分别为0.239±0.037、0.235±0.006比0.199±0.007,P＜0.01)且部分呈线性分布于闰盘,C组和D组之间差异无统计学意义(P＞0.05).结论心肌组织中Ang Ⅱ含量长期升高可能参与心房纤维化的形成和Cx40重构,缬沙坦与U0126通过抑制ERK通路的不同位点,在改善心房纤维化程度和Cx40重构方面发挥相似的作用.
목적 탐토힐사탄여U0126대혈관긴장소Ⅱ(angiotensin Ⅱ,Ang Ⅱ)유도적대서심방섬유화화봉극련접단백40( connexin 40,Cx40)중구적영향.방법 장32지웅성SD대서수궤분위공백대조조(A조)、염산이병기신상선소( isopreterenol,ISO)+이갑기아풍(DMSO)조(B조)、ISO+U0126조(C조,U0126용우DMSO중)、ISO+힐사탄+DMSO조(D조).급약28 d후처사대서취심기조직,방사면역법측Ang Ⅱ함량;HE화Masson염색법관찰섬유화정도즉효원용적분수(CVF);면역조화법측정린산화사렬원활화단백격매격매1/2(P-MEK1/2)、린산화세포외신호조절격매1/2( P-ERK1/2)이급Cx40적표체.결과 B、C、D조중Ang Ⅱ함량교A조명현승고[분별위(368.243±6.283)ng/L、(357.175±5.944) ng/L、(359.908±2.496) ng/L비(250.380±4.261) ng/L,P＜0.01];A조CVF(9.025 ±0.456)％,현시무심방섬유화;C조화D조교B조심방섬유화정도명현감약[CVF분별위(10.260±0.525)％、(10.238 ±0.524)％비(78.710±1.587)％,P＜0.01],C조화D조지간차이무통계학의의(P＞0.05);B조교A조중P-MEK1/2(0.311±0.007비0.203±0.009,P＜0.01)화P-ERK1/2함량명현증가(0.259±0.003비0.173±0.006,P＜0.01),이C조화D조중함량교B조명현감소(P-MEK1/2분별위0.212±0.004、0.213±0.005비0.311±0.007,P＜0.01;P-ERK1/2분별위0.178±0.004、0.175 ±0.007비0.259±0.003,P＜0.01),C조화D조지간차이무통계학의의(P＞0.05);B조교A조Cx40함량명현감소(0.199±0.007비0.241±0.004,P＜0.01)차분포문란,C조화D조중함량교B조감소정도명현감경(분별위0.239±0.037、0.235±0.006비0.199±0.007,P＜0.01)차부분정선성분포우윤반,C조화D조지간차이무통계학의의(P＞0.05).결론 심기조직중Ang Ⅱ함량장기승고가능삼여심방섬유화적형성화Cx40중구,힐사탄여U0126통과억제ERK통로적불동위점,재개선심방섬유화정도화Cx40중구방면발휘상사적작용.
Objective To explore the effects of valsartan and MEK1/2 inhibitor U0126 on atrial fibrosis and connexin40 (Cx40) remodeling in rats treated with isopreterenol (ISO).Methods 32 male SD rats were randomly divided into control group (A),ISO (5 mg · kg-1 · d-1 for7 days) + DMSO group (B),ISO + U0126 (0.5 mg · kg-1 · d-1 for 28 days) group (C,U0126 was dissolved in DMSO),ISO +valsartan (30 mg · kg-1 · d-1 for 28 days) + DMSO group (D).Rats were sacrificed after 28 days.The AngⅡ content in myocardial tissue was measured by radioimmunoassay,P-MEK1/2,P-ERK1/2 and Cx40 was detected by immunohistochemistry,atrial fibrosis was determined on HE and Masson stained heart sections.Results The content of Ang Ⅱ was significantly higher in group B,C and D compared with group A [ (368.243 ±6.283 ) ng/L,( 357.175 ± 5.944 ) ng/L,( 359.908 ± 2.496 ) ng/L vs ( 250.380 ± 4.261 )ng/L,P ＜0.01 ] ; the degree of atrial fibrosis was significantly lower in group C and D compared with group B [CVF(10.260 ±0.525)％,(10.238 ±0.524)％ vs (78.710 ± 1.587)％,P＜0.01 ] while there was no atrial fibrosis in group A [ CVF(9.025 ± 0.456)％ ] ; the expression of P-MEK1/2 and P-ERK1/2 was significantly upregulated in group B compared with group A ( P-MEK1/2:0.311 ± 0.007 vs 0.203 ± 0.009,P ＜0.01 ; P-ERK1/2:0.259 ±0.003 vs 0.173 ±0.006,P ＜0.01 ) and significantly lower in group C and D compared with group B (P-MEK1/2:0.212 ± 0.004,0.213 ± 0.005 vs 0.311 ± 0.007,P ＜0.01,P-ERK1/2:0.178 ±0.004,0.175 ±0.007 vs 0.259 ±0.003,P ＜0.01 ).The content of Cx40 was obviously reduced and the distribution of Cx40 was disordered in group B compared with A (0.199 ±0.007 vs 0.241 ±0.004,P＜0.01) which could be partly reversed in group C and D ( 0.239 ±0.037,0.235 ±0.006 vs 0.199 ± 0.007,P ＜ 0.01 ).All parameters in group C and D were similar ( P ＞ 0.05 ).Conclusion The chronically elevated Ang Ⅱ content in myocardium may be related to atrial fibrosis and Cx40 remodeling in this model,valsartan and U0126 were equivalent on attenuating atrial fibrosis and Cx40 remodeling by inhibiting ERK pathways at different levels.