miR-21通过细胞凋亡途径调节脓毒症大鼠早期外周血Th17/Treg介导的免疫平衡

IF 3.4 Q2 BIOCHEMICAL RESEARCH METHODS
Cheng Liu, Qi Zou
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引用次数: 0

摘要

目的探讨miR-21在脓毒症中调节Th17/Treg细胞分化和功能的机制。方法采用盲肠结扎穿刺法建立大鼠败血症模型。然后,通过脂质体用miR-21模拟物或抑制剂转染一些脓毒症大鼠。48 小时后,分离脓毒症大鼠的淋巴细胞和血浆进行进一步的实验检测。用聚合酶链式反应(PCR)检测淋巴细胞中miR-21的表达;通过流式细胞术计数Th17/Treg细胞的分化;用末端脱氧核苷酸转移酶介导的dUTP生物素缺口末端标记法(TUNEL)观察淋巴细胞凋亡。用蛋白质印迹法检测胱天蛋白酶3/9蛋白;用酶联免疫吸附试验(ELISA)检测IL-10和IL-17。结果与败血症组(SP组)相比,转染miR-21的败血症组Th17细胞显著增加,Treg细胞显著减少,淋巴细胞凋亡率显著降低,半胱氨酸天冬氨酸蛋白酶3/9 mRNA和蛋白显著降低,IL-17降低,IL-10升高 + miR-21模拟组)。转染miR-21抑制剂后,结果几乎与SP相反 + miR-21模拟组。结论miR-21通过caspase途径调控脓毒症Th17/Treg细胞的分化和功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
miR-21 Regulates Immune Balance Mediated by Th17/Treg in Peripheral Blood of Septic Rats during the Early Phase through Apoptosis Pathway
Objective To study the mechanism by which miR-21 regulates the differentiation and function of Th17/Treg cells in sepsis. Methods A rat model with sepsis was made by cecal ligation and puncture (CLP). Then, some of the septic rats were transfected with miR-21 mimic or inhibitor by liposome. At 48 hours, lymphocytes and plasma from septic rats were isolated for further experimental detection. The expression of miR-21 in lymphocytes was detected by Polymerase Chain Reaction (PCR); the differentiation of Th17/Treg cells was counted by flow cytometry; lymphocyte apoptosis was observed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. The caspase-3/9 proteins were tested by Western blot; IL-10 and IL-17 were detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the sepsis group (SP group), the Th17 cells increased significantly, the Treg cells decreased significantly, the apoptosis rate of lymphocytes decreased significantly, the mRNA and proteins of caspase-3/9 decreased significantly, the IL-17 decreased, and the IL-10 increased in the sepsis group transfected with miR-21 (SP + miR-21 mimic group). After transfection of miR-21 inhibitor, the results were almost opposite to those of SP + miR-21 mimic group. Conclusions The differentiation and function of Th17/Treg cells were regulated by miR-21 in sepsis through caspase pathway.
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来源期刊
Biochemistry Research International
Biochemistry Research International BIOCHEMICAL RESEARCH METHODS-
CiteScore
6.30
自引率
0.00%
发文量
27
审稿时长
14 weeks
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