腺相关病毒 9 介导的 SOCS3 靶向 RNA 干扰减轻大鼠舒张性心力衰竭。
Adeno-associated virus 9-mediated RNA interference targeting SOCS3 alleviates diastolic heart failure in rats.
机构信息
Department of Cardiac Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China.
Department of Cardiac Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China.
出版信息
Gene. 2019 May 20;697:11-18. doi: 10.1016/j.gene.2019.01.044. Epub 2019 Feb 11.
OBJECTIVE
To explore the effect of adeno-associated virus 9-mediated RNA interference targeting SOCS3 (AAV9-SOCS3 siRNA) on the treatment of diastolic heart failure (DHF).
METHOD
A rat DHF model was established, and cardiac function and hemodynamic changes were measured. HE, Sirius red and TUNEL staining were applied to observe the pathological changes in the myocardium. Immunoblotting and immunohistochemical staining were utilized to detect SOCS3 expression. The expression levels of various factors, including fibrosis-related factors (collagen I, collagen II, α-SMA and TGF-β), inflammatory-related factors (IL-1β, IL-6, TNF-α, p-p65 and ICAM-1) and factors related to the JAK/STAT signal pathway were analyzed by immunoblotting and/or qPCR. The serum levels of IL-1β, IL-6, and TNF-α were measured using ELISA.
RESULTS
SOCS3 expression was significantly downregulated in the DHF rat model by SOCS3 siRNA delivery. In the successfully established DHF rat model, cardiac function was clearly decreased, and cardiomyocyte apoptosis and myocardial fibrosis were significantly increased. These changes were ameliorated by treatment with AAV9-SOCS3 siRNA. The expression levels of p-JAK2 and p-STAT3 were significantly upregulated in the AAV9-SOCS3 siRNA group compared with the sham and AAV9-siRNA control groups, indicating that SOCS3 is a negative regulator of this signaling pathway. The expression levels of collagen I/III, α-SMA and TGF-β were also decreased at both the mRNA and protein levels. In addition, the serum and myocardial tissue expression levels of inflammatory-related factors, such as IL-6, IL-1β, and TNF-α, were also reduced by the administration of AAV9-SOCS3 siRNA compared with the AAV9-siRNA control.
CONCLUSIONS
SOCS3 gene silencing by AAV9-SOCS3 siRNA administration in a DHF rat model significantly reduced myocardial fibrosis and the inflammatory response and improved heart function. Therefore, this treatment is a potential therapeutic method for treating DHF.
目的
探讨腺相关病毒 9 介导的 SOCS3 靶向 RNA 干扰（AAV9-SOCS3 siRNA）对舒张性心力衰竭（DHF）治疗的影响。
方法
建立大鼠 DHF 模型，检测心功能和血流动力学变化，HE、天狼猩红和 TUNEL 染色观察心肌病理变化，免疫印迹和免疫组化检测 SOCS3 表达，免疫印迹和/或 qPCR 分析纤维化相关因子（胶原 I、胶原 II、α-SMA 和 TGF-β）、炎症相关因子（IL-1β、IL-6、TNF-α、p-p65 和 ICAM-1）和 JAK/STAT 信号通路相关因子的表达水平，ELISA 检测血清中 IL-1β、IL-6 和 TNF-α 的含量。
结果
通过 SOCS3 siRNA 转染，DHF 大鼠模型中 SOCS3 表达明显下调。在成功建立的 DHF 大鼠模型中，心脏功能明显降低，心肌细胞凋亡和心肌纤维化明显增加，用 AAV9-SOCS3 siRNA 治疗后得到改善。与 sham 和 AAV9-siRNA 对照组相比，AAV9-SOCS3 siRNA 组中 p-JAK2 和 p-STAT3 的表达明显上调，表明 SOCS3 是该信号通路的负调节剂。胶原 I/III、α-SMA 和 TGF-β 的表达水平在 mRNA 和蛋白水平均降低。此外，与 AAV9-siRNA 对照组相比，AAV9-SOCS3 siRNA 给药后，血清和心肌组织中炎症相关因子（如 IL-6、IL-1β 和 TNF-α）的表达水平也降低。
结论
在 DHF 大鼠模型中，AAV9-SOCS3 siRNA 给药沉默 SOCS3 基因可显著减轻心肌纤维化和炎症反应，改善心功能，因此，这种治疗方法可能是治疗 DHF 的一种潜在治疗方法。
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