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miR-21在早期通过凋亡途径调节脓毒症大鼠外周血中由Th17/Treg介导的免疫平衡。

miR-21 Regulates Immune Balance Mediated by Th17/Treg in Peripheral Blood of Septic Rats during the Early Phase through Apoptosis Pathway.

作者信息

Liu Cheng, Zou Qi

机构信息

Department of Critical Care Medicine, The First Affiliated Hospital of Bengbu Medical College, Anhui, Bengbu 233004, China.

出版信息

Biochem Res Int. 2022 Apr 27;2022:9948229. doi: 10.1155/2022/9948229. eCollection 2022.

DOI:10.1155/2022/9948229
PMID:35528843
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9068307/
Abstract

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.

摘要

目的

研究miR-21调控脓毒症中Th17/Treg细胞分化及功能的机制。

方法

采用盲肠结扎穿刺术（CLP）制备大鼠脓毒症模型。然后，部分脓毒症大鼠通过脂质体转染miR-21模拟物或抑制剂。48小时后，分离脓毒症大鼠的淋巴细胞和血浆用于进一步实验检测。采用聚合酶链反应（PCR）检测淋巴细胞中miR-21的表达；通过流式细胞术计数Th17/Treg细胞的分化情况；采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）检测淋巴细胞凋亡。通过蛋白质印迹法检测caspase-3/9蛋白；采用酶联免疫吸附测定法（ELISA）检测IL-10和IL-17。

结果

与脓毒症组（SP组）相比，转染miR-21的脓毒症组（SP + miR-21模拟物组）中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，caspase-3/9的mRNA和蛋白显著减少，IL-17减少，IL-10增加。转染miR-21抑制剂后，结果与SP + miR-21模拟物组几乎相反。

结论

脓毒症中miR-21通过caspase途径调控Th17/Treg细胞的分化及功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/f3f141644493/BRI2022-9948229.008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/f3f141644493/BRI2022-9948229.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/13f3497ba268/BRI2022-9948229.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/f8c517a9a078/BRI2022-9948229.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/2678501b5f75/BRI2022-9948229.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/e861951c2f1c/BRI2022-9948229.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/9fa8d0dab294/BRI2022-9948229.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/a37b4579c2fd/BRI2022-9948229.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/0680693d9dc4/BRI2022-9948229.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7e16/9068307/f3f141644493/BRI2022-9948229.008.jpg

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