Department of Critical Care Medicine, The first Affiliated Hospital of Bengbu Medical College, Anhui, 233004, Bengbu, People's Republic of China.
Department of Pediatrics, The first Affiliated Hospital of Bengbu Medical College, Anhui, 233004, Bengbu, People's Republic of China.
Inflammation. 2020 Dec;43(6):2287-2300. doi: 10.1007/s10753-020-01298-7.
To observe the inflammatory response, differentiation of Th17/Treg subsets and apoptosis of lymphocytes, by regulating miR-126 in lymphocytes of septic rats. After using cecal ligation and puncture to establish sepsis model, miR-126 mimic and miR-126 inhibitor were used to transfect lymphocytes of septic rats in vitro and in vivo. ELISA was used to detect TNF-α, IL-6, IL-17, and IL-10, the differentiation of Th17 and Treg was measured by flow cytometry, and apoptosis of lymphocytes was observed by fluorescence microscope; the changes of caspase signaling pathway were detected by immunofluorescence, PCR, and Western blotting. The result show that the expression of miR-126 increased in sepsis. After overexpression of miR-126, the release of TNF-α, IL-6, and IL-17 decreased; the release of IL-10 increased; T lymphocyte subsets differentiated toward Treg; caspase signaling pathway weakened; and lymphocyte of apoptosis decreased compared with sepsis group. While, after inhibition of miR-126, the release of TNF-α, IL-6, and IL-17 increased; the release of IL-10 decreased; T lymphocyte subsets differentiated toward TH17; caspase signaling pathway enhanced; and lymphocyte of apoptosis increased compared with sepsis group. Taken together, regulation of miR-126 can alter the inflammatory response, differentiation of T lymphocyte subsets, and apoptosis of lymphocytes in septic rats.
观察通过调节脓毒症大鼠淋巴细胞中的 miR-126 对 Th17/Treg 亚群分化和淋巴细胞凋亡的影响。采用盲肠结扎穿孔法建立脓毒症模型,体外和体内转染脓毒症大鼠淋巴细胞 miR-126 模拟物和 miR-126 抑制剂。ELISA 法检测 TNF-α、IL-6、IL-17、IL-10,流式细胞术检测 Th17 和 Treg 分化,荧光显微镜观察淋巴细胞凋亡;免疫荧光、PCR 和 Western blot 检测 caspase 信号通路变化。结果显示,脓毒症时 miR-126 表达增加。miR-126 过表达后,TNF-α、IL-6、IL-17 释放减少,IL-10 释放增加,T 淋巴细胞亚群向 Treg 分化,caspase 信号通路减弱,凋亡的淋巴细胞减少,与脓毒症组比较差异有统计学意义。而 miR-126 抑制后,TNF-α、IL-6、IL-17 释放增加,IL-10 释放减少,T 淋巴细胞亚群向 TH17 分化,caspase 信号通路增强,凋亡的淋巴细胞增加,与脓毒症组比较差异有统计学意义。综上,调节 miR-126 可以改变脓毒症大鼠的炎症反应、T 淋巴细胞亚群分化和淋巴细胞凋亡。