Department of Urology, China-Japan Union Hospital of Jilin University, No. 126, Xiantai Street, Changchun, 130033, Jilin Province, People's Republic of China.
Int Urogynecol J. 2021 May;32(5):1253-1263. doi: 10.1007/s00192-020-04593-x. Epub 2021 Jan 8.
As a notable chronic disorder, the incidence of interstitial cystitis (IC) has been documented to have increased among the female population with activity in microRNA-495 (miR-495) implicated in this disease. The current study was aimed at elucidating the effects associated with miR-495 on the inflammatory response and bladder fibrosis in rats with ulcerative IC via the JAK-STAT pathway by targeting JAK3.
Ulcerative IC rat models were established. The targeting relationship between JAK3 and miR-495 was evaluated using luciferase reporter assay. After gain- or loss-of-function assays, mast-cell infiltration was assessed using toluidine blue staining, bladder fibrosis using Masson staining, and NO content using nitrate reductase method. JAK3 protein expression was detected by immunohistochemistry, JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, JAK1, JAK2, p-STAT1, and p-STAT3 expression by RT-qPCR and Western blot analysis, and serum IL-6, IL-8, IL-10, IL-17, and TNF-α levels in rats by ELISA.
Following transfection of overexpressed miR-495 or siRNA-JAK3, a diminished degree of mast-cell infiltration, number of mast cells, bladder fibrosis, NO content, JAK3-positive expression, mRNA expression of JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, protein expression of JAK1, JAK2, JAK3, p-STAT1, p-STAT3, and expression of IL-6, IL-8, IL-10, IL-17, and TNF-α were identified.
Collectively, our key findings provide evidence supporting the notion that the overexpression of miR-495 ameliorates inflammatory response and bladder fibrosis in ulcerative IC rat models via inactivation of the JAK-STAT signaling pathway by inhibiting JAK3.
作为一种显著的慢性疾病，间质性膀胱炎 (IC) 的发病率已被记录在女性人群中有所增加，其中 microRNA-495 (miR-495) 的活性与这种疾病有关。本研究旨在通过靶向 JAK3 阐明 miR-495 通过 JAK-STAT 通路对溃疡性 IC 大鼠炎症反应和膀胱纤维化的影响。
建立溃疡性 IC 大鼠模型。通过荧光素酶报告实验评估 JAK3 和 miR-495 之间的靶向关系。在获得或丧失功能后，使用甲苯胺蓝染色评估肥大细胞浸润，使用 Masson 染色评估膀胱纤维化，使用硝酸盐还原酶法评估 NO 含量。通过免疫组织化学检测 JAK3 蛋白表达，通过 RT-qPCR 和 Western blot 分析检测 JAK3、STAT1、STAT3、TGFβ-1、Col-I、Col-III、JAK1、JAK2、p-STAT1 和 p-STAT3 的表达，通过 ELISA 检测大鼠血清中 IL-6、IL-8、IL-10、IL-17 和 TNF-α的水平。
转染过表达 miR-495 或 siRNA-JAK3 后，肥大细胞浸润程度、肥大细胞数量、膀胱纤维化、NO 含量、JAK3 阳性表达、JAK3、STAT1、STAT3、TGFβ-1、Col-I、Col-III、JAK1、JAK2、JAK3、p-STAT1、p-STAT3 的 mRNA 表达以及 IL-6、IL-8、IL-10、IL-17 和 TNF-α的蛋白表达均降低。
综上所述，我们的主要发现为 miR-495 通过抑制 JAK3 使 JAK-STAT 信号通路失活，从而改善溃疡性 IC 大鼠模型中的炎症反应和膀胱纤维化的观点提供了证据。
