Xbp1s-Ddit3 promotes MCT-induced pulmonary hypertension.
Xbp1s-Ddit3 promotes MCT-induced pulmonary hypertension.
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DOI：
10.1042/cs20210612
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发表时间：
2021-11-12
期刊：
影响因子：
--
通讯作者：
Liu X
中科院分区：
文献类型：
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作者：
Jiang H;Ding D;He Y;Li X;Xu Y;Liu X
肺动脉高压（PH）是一种以血管重塑为特征的危及生命的疾病。探索新的治疗靶点迫在眉睫。本研究旨在探讨剪接的x-box结合蛋白1（xbp 1 s）作为内质网应激（ERS）的重要组成部分，是否以及如何参与PH的发病。将40只雄性SD大鼠随机分为4组：对照组、野百合碱（MCT）组、MCT+AAV-CTL组（对照组）和MCT+ AAV-xbp 1 s组。发现MCT组肺组织中xbp 1 s蛋白水平升高。腹腔内注射携带xbp 1 s shRNA的1型腺相关病毒（AAV-xbp 1 s）可有效抑制MCT引起的右心室收缩压（RVSP）、总肺阻力（TPR）、右心室肥厚和远端肺小动脉中膜厚度的增加。MCT+ AAV-xbp 1 s组肺小动脉增殖细胞核抗原（PCNA）和Ki 67阳性染色率异常升高，末端脱氧核苷酸转移酶（TdT）介导的dUTP缺口末端标记（TUNEL）阳性染色率降低。对于机制探索，在STRING数据库上进行蛋白质网络的生物信息学预测，并通过qRT-PCR、Western印迹和免疫共沉淀（Co-IP）进行进一步验证。DNA损伤诱导转录本3（Ddit 3）被鉴定为与xbp 1 s相互作用的下游蛋白。Ddit 3的过表达恢复了由xbp 1 s沉默引起的增殖、迁移和细胞活力的下降。在动物模型中，Ddit 3的蛋白水平也与xbp 1 s高度一致。综上所述，我们的研究表明，xbp 1 s-Ddit 3可能是一个潜在的目标，以干扰血管重塑的PH。
Pulmonary hypertension (PH) is a life-threatening disease characterized by vascular remodeling. Exploring new therapy target is urgent. The purpose of the present study is to investigate whether and how spliced x-box binding protein 1 (xbp1s), a key component of endoplasmic reticulum stress (ERS), contributes to the pathogenesis of PH. Forty male SD rats were randomly assigned to four groups: Control, Monocrotaline (MCT), MCT+AAV-CTL (control), and MCT+AAV-xbp1s. The xbp1s protein levels were found to be elevated in lung tissues of the MCT group. Intratracheal injection of adeno-associated virus serotype 1 carrying xbp1s shRNA (AAV-xbp1s) to knock down the expression of xbp1s effectively ameliorated the MCT-induced elevation of right ventricular systolic pressure (RVSP), total pulmonary resistance (TPR), right ventricular hypertrophy and medial wall thickness of muscularized distal pulmonary arterioles. The abnormally increased positive staining rates of proliferating cell nuclear antigen (PCNA) and Ki67 and decreased positive staining rates of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) in pulmonary arterioles were also reversed in the MCT+AAV-xbp1s group. For mechanistic exploration, bioinformatics prediction of the protein network was performed on the STRING database, and further verification was performed by qRT-PCR, Western blots and co-immunoprecipitation (Co-IP). DNA damage-inducible transcript 3 (Ddit3) was identified as a downstream protein that interacted with xbp1s. Overexpression of Ddit3 restored the decreased proliferation, migration and cell viability caused by silencing of xbp1s. The protein level of Ddit3 was also highly consistent with xbp1s in the animal model. Taken together, our study demonstrated that xbp1s-Ddit3 may be a potential target to interfere with vascular remodeling in PH.