CAJ | 학술논문

目的研究谷氨酰胺(Gln)与缬沙坦对大鼠心房纤维化及缝隙连接蛋白43(Cx43)重构的影响及其可能的作用机制.方法将SD雄性大鼠(160±20)9 32只随机等分为对照组(A组)、盐酸异丙肾上腺素(ISO)(5 mg·kg-1·d-1)(B组)、ISO(5 mg·kg-1·d-1)+丙氨酰谷酰胺注射液(Ala-Gln)(0.75 mg·kg-1·d-1)(C组)和ISO(5 mg·kg-1·d-1)+缬沙坦(30 mg·kg-1·d-1)(D组).各组1次/d给予相关试剂,ISO连续给药7d,而后C、D两组大鼠继续给予相关试剂至28 d后处死.心肌组织中血管紧张素Ⅱ(Ang Ⅱ)含量采用放免法检测；通过H-E染色观察心肌纤维化情况；Masson染色计算胶原容积分数(CVF)并作为心肌纤维化程度的量化指标；免疫组化SP法半定量测定心肌组织中热休克蛋白70(HSP70)、磷酸化c-Jun氨基末端激酶1/2/3(p-JNK1/2/3)、c-Jun及Cx43蛋白的表达.结果 Ang Ⅱ在B、C、D组中含量较A组显著升高且差异有统计学意义(P＜0.01)；B组与A、C、D组相比出现明显的心房纤维化(P＜0.01)且心肌组织中p-JNK1/2/3和c-Jun的含量较高,差异有统计学意义(P＜0.01),C、D组与A组相比心房纤维化程度及心肌组织中p-JNK1/2/3和c-Jun的含量差异无统计学意义(P＞0.05)；与其他各组相比,C组中HSP70的含量明显升高,且差异有统计学意义(P＜0.01),A、B、D组中HSP70的含量相互间差异均无统计学意义(P＞0.05)；B组中Cx43含量较A组减少(P＜0.01)且分布无规律性,侧面化分布相对增多.而C、D组中Cx43含量与A组比较差异无统计学意义(P＞0.05)且部分呈线性分布于心肌细胞闰盘处；C、D两组中Cx43含量比较差异均无统计学意义(P＞0.05).结论谷氨酰胺与缬沙坦均可减轻Ang Ⅱ诱发的大鼠心房纤维化及Cx43重构,其机制可能与降低p-JNK1/2/3和c-Jun的表达,抑制JNK信号通路的不同位点有关,在改善大鼠心房纤维化程度和Cx43重构方面起到类似的作用.
목적 연구곡안선알(Gln)여힐사탄대대서심방섬유화급봉극련접단백43(Cx43)중구적영향급기가능적작용궤제.방법 장SD웅성대서(160±20)9 32지수궤등분위대조조(A조)、염산이병신상선소(ISO)(5 mg·kg-1·d-1)(B조)、ISO(5 mg·kg-1·d-1)+병안선곡선알주사액(Ala-Gln)(0.75 mg·kg-1·d-1)(C조)화ISO(5 mg·kg-1·d-1)+힐사탄(30 mg·kg-1·d-1)(D조).각조1차/d급여상관시제,ISO련속급약7d,이후C、D량조대서계속급여상관시제지28 d후처사.심기조직중혈관긴장소Ⅱ(Ang Ⅱ)함량채용방면법검측；통과H-E염색관찰심기섬유화정황；Masson염색계산효원용적분수(CVF)병작위심기섬유화정도적양화지표；면역조화SP법반정량측정심기조직중열휴극단백70(HSP70)、린산화c-Jun안기말단격매1/2/3(p-JNK1/2/3)、c-Jun급Cx43단백적표체.결과 Ang Ⅱ재B、C、D조중함량교A조현저승고차차이유통계학의의(P＜0.01)；B조여A、C、D조상비출현명현적심방섬유화(P＜0.01)차심기조직중p-JNK1/2/3화c-Jun적함량교고,차이유통계학의의(P＜0.01),C、D조여A조상비심방섬유화정도급심기조직중p-JNK1/2/3화c-Jun적함량차이무통계학의의(P＞0.05)；여기타각조상비,C조중HSP70적함량명현승고,차차이유통계학의의(P＜0.01),A、B、D조중HSP70적함량상호간차이균무통계학의의(P＞0.05)；B조중Cx43함량교A조감소(P＜0.01)차분포무규률성,측면화분포상대증다.이C、D조중Cx43함량여A조비교차이무통계학의의(P＞0.05)차부분정선성분포우심기세포윤반처；C、D량조중Cx43함량비교차이균무통계학의의(P＞0.05).결론 곡안선알여힐사탄균가감경Ang Ⅱ유발적대서심방섬유화급Cx43중구,기궤제가능여강저p-JNK1/2/3화c-Jun적표체,억제JNK신호통로적불동위점유관,재개선대서심방섬유화정도화Cx43중구방면기도유사적작용.
Objective To study the effects of glutamine and valsartan on atrial fibrosis and connexin43 remodeling in rats induced by isoprenaline(ISO)and the possible mechanisms.Methods Thirty-two male SD rats (160±20)g were randomly divided into four groups (n =8):control group (A),ISO (5 mg · kg-1 · d-1)group (B),and ISO(5 mg · kg-1 · d-1)+Ala-Gln(0.75 mg · kg-1 · d-1) group (C),ISO(5 mg · kg-1 · d-1)+valsartan(30 mg · kg-1 · d-1)group(D).Each group was treated with the corresponding reagent once a day,ISO administration for seven days,group C and D continued to be treated with corresponding drugs for 28 days,all rats were killed after 28 days.The Ang Ⅱ content was measured by radioimmunoassay ; myocardial fibrosis was analyzed by H-E staining;Collagen volume fractions were quantified by Masson staining and as the indicators of atrial fibrosis; The expressions of HSP70,p-JNK1/2/3,c-Jun and Cx43 were semi-quantified with immunohistochemical method.Results Compared with the A group,the contents of Ang Ⅱ in group B、C、and D were significantly increased(P＜0.01,respectively).No obvious atrial fibrosis was observed in group A,and there was evident atrial fibrosis in group B,and was much higher than that in group C、group D(P＜0.01,respectively).The expressions of p-JNK1/2/3 and c-Jun in group B were significantly increased(P＜0.01),while the expressions of p-JNK1/2/3 and c-Jun in the group A 、C and D were not obvious change(P＞0.05),The content of HSP70 in group C was significantly increased,and the difference was statistically was significant higher than that in other groups (P＜0.01,respectively).There was no significant difference in contents of HSP70 among the groups of A、B and D.The content of Cx43 in group B was obvious reduced(P＜0.01)with irregular distribution,relatively increased in the side of the myocardial cells.While the content of Cx43 in group C and D had no obvious change compared with that in A group(P＞0.05).The Cx43 partly linearly distributed in myocardial cell intercalated disc,and the content of Cx43 in group C and D showed no significant difference (P＞0.05).Conclusion Glutamine and valsartan can reduce the atrial fibrosis and connexin 43 remodeling in rats induced by isoprenaline.The mechanism may be relate to inhibit JNK pathways at different levels.