Xie Xiaoran, Liu Peng, Wu Hao, Li Huan, Tang Yu, Chen Xiong, Xu Canxia, Liu Xiaoming, Dai Guoyu
Department of Gastroenterology, The Third Xiangya Hospital of Central South University, Changsha, China.
Hunan Provincial Key Laboratory of Uncontrollable Inflammation and Tumor, The Third Xiangya Hospital of Central South University, Changsha, China.
Ann Transl Med. 2022 Apr;10(7):413. doi: 10.21037/atm-22-944.
The pathogenesis of Crohn's disease (CD) is unknown; however, angiogenesis is known to play an important role in the disease. The present research suggests that microRNA-21 (miR-21) may play a positive regulatory role in disordered angiogenesis in CD.
C57 wild-type mice were divided into 6 groups. On day 0, all mice in the 2,4,6-trinitrobenzenesulfonic acid (TNBS) group were given an enema at the concentration of TNBS 100 mg/kg mouse body weight (solvent 50% alcohol). In the control group, the enema was performed with 50% alcohol. On day 0, 2, 4, and 6, the mice of the agomir-21 + TNBS group and the agomir control + TNBS group were injected with 200 µL, 5 nmol agomir-21 or agomir control [dissolved in ribonuclease (RNase)-free water] by tail vein injection, while the antagomir-21 + TNBS group and the antagomir control + TNBS group were injected with 200 µL, 20 nmol antagomir-21 or antagomir control (dissolved in RNase-free water). The body weight and disease activity index (DAI) score were recorded daily. The colons were obtained to assess macro and microscopic colon damage. The inferior vena cava and the accompanying abdominal aorta were chosen to detect the protein expression of the phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/serine/threonine kinase (AKT)/vascular endothelial growth factor (VEGF) axis through western blotting. Serum interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The distribution and expression of neovascularization were demonstrated by cluster of differentiation 31 (CD31) immunohistochemistry.
Compared with the only-TNBS group, the agomir-21 + TNBS group showed significantly severer colitis symptoms and more abnormal vascular hyperplasia, while the antagomir-21 + TNBS group showed symptom relief and reduced vascular hyperplasia. In addition, agomir-21 obviously inhibited the expression of PTEN and activated the PI3K/AKT/VEGF pathway in mice induced by TNBS, while antagomir-21 effectively antagonized this effect.
miR-21 can promote the progression of colitis in mice induced by TNBS and aggravate the disordered angiogenesis by regulating the PTEN/PI3K/AKT axis. Intravenous injection of miR-21 antagonists can effectively relieve the symptoms of colitis and inhibit colonic angiogenesis.
克罗恩病（CD）的发病机制尚不清楚；然而，已知血管生成在该疾病中起重要作用。目前的研究表明，微小RNA-21（miR-21）可能在CD紊乱的血管生成中发挥正向调节作用。
将C57野生型小鼠分为6组。在第0天，2,4,6-三硝基苯磺酸（TNBS）组的所有小鼠均以100 mg/kg小鼠体重的TNBS浓度（溶剂为50%乙醇）进行灌肠。对照组用50%乙醇进行灌肠。在第0、2、4和6天，agomir-21 + TNBS组和agomir对照 + TNBS组的小鼠通过尾静脉注射200 μL、5 nmol的agomir-21或agomir对照[溶解于无核糖核酸酶（RNase）的水中]，而antagomir-21 + TNBS组和antagomir对照 + TNBS组的小鼠则注射200 μL、20 nmol的antagomir-21或antagomir对照（溶解于无RNase的水中）。每天记录体重和疾病活动指数（DAI）评分。获取结肠以评估大体和显微镜下的结肠损伤。选择下腔静脉及其伴随的腹主动脉，通过蛋白质印迹法检测磷酸酶和张力蛋白同源物（PTEN）/磷脂酰肌醇3激酶（PI3K）/丝氨酸/苏氨酸激酶（AKT）/血管内皮生长因子（VEGF）轴的蛋白表达。通过酶联免疫吸附测定（ELISA）检测血清白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）。通过分化簇31（CD31）免疫组织化学显示新生血管的分布和表达。
与仅TNBS组相比，agomir-21 + TNBS组显示出明显更严重的结肠炎症状和更多异常的血管增生，而antagomir-21 + TNBS组显示症状缓解且血管增生减少。此外，agomir-21明显抑制TNBS诱导的小鼠中PTEN的表达并激活PI3K/AKT/VEGF途径，而antagomir-21有效拮抗了这种作用。
miR-21可促进TNBS诱导的小鼠结肠炎进展，并通过调节PTEN/PI3K/AKT轴加重紊乱的血管生成。静脉注射miR-21拮抗剂可有效缓解结肠炎症状并抑制结肠血管生成。
