RhoF介导的Th17极化在急性胰腺炎发生中的作用及机制

doi:10.3969/j.issn.1006-5725.2024.10.004

摘要/Abstract

摘要：

目的探讨Rho相关GTP结合蛋白F（RhoF）介导的Th17极化在重度急性胰腺炎（SAP）发生中的作用及机制研究。方法将RhoF转基因小鼠随机分为3组：对照组（n = 10）、SAP组（n = 10）和SAP+Y-27632组（n = 10）。另将WT小鼠随机分为3组：对照组（n = 10）、SAP组（n = 10）和SAP+Y-27632组（n = 10）。除对照组外，其他组建立SAP模型。SAP+Y-27632组在模型建立后，通过尾静脉输注Y-27632。分离小鼠血液样本中的T细胞，用于RhoF、p-MYPT1蛋白检测和ROCK活性测定，并采用流式细胞仪分析产生IL-17的淋巴CD4⁺T细胞的百分比。结果与对照组相比，SAP组小鼠T细胞中RhoF、p-MYPT1表达量增加（P < 0.01），并且RhoF的水平与p-MYPT1的水平密切相关（P < 0.05）。与WT组相比，RhoF组小鼠T细胞的RhoF蛋白水平的表达增加约30%，并且CD4⁺ T细胞自发产生IL-17的水平显著增加（P < 0.01）。与WT小鼠相比，RhoF转基因小鼠胰腺损伤的组织学评分，T细胞的RhoF、p-MYPT1表达量，IL-17⁺ T细胞数量和血清IL-17水平明显增加（P < 0.05）。SAP+Y-27632组的组织学评分，T细胞的RhoF、p-MYPT1表达量，IL-17⁺ T细胞数量和血清IL-17水平显著低于SAP组（P < 0.05）。结论 RhoF/ROCK信号通路介导的Th17细胞极化参与SAP的发病机制。

关键词: Rho相关GTP结合蛋白F, Th17细胞, 重症急性胰腺炎

Abstract:

Objective To investigate the role and mechanism of Rho-related GTP-binding protein F （RhoF）-mediated Th17 polarization in the development of severe acute pancreatitis （SAP）. Methods RhoF transgenic mice were randomly divided into control group （n = 10）， SAP group （n = 10） and SAP+Y-27632 group （n = 10）， and WT mice were randomly divided into control group （n = 10）， SAP group （n = 10） and SAP+Y-27632 group （n = 10）. SAP models were established in all groups except the control group. The SAP+Y-27632 group was infused with Y-27632 via tail vein after model establishment. T cells in mouse blood samples were isolated for RhoF， p-MYPT1 protein detection and ROCK activity determination， and the percentage of lymphoid CD4⁺T cells producing IL-17 was analyzed by flow cytometry. Results The expression of RhoF and p-MYPT1 was increased in T cells in the SAP group compared with that in the control group （P < 0.01）， and the level of RhoF was closely correlated with that of p-MYPT1 （P = 0.018）. The expression of RhoF protein level in T cells of mice in the RhoF group increased by approximately 30% compared to that in the WT group， and the level of spontaneous IL-17 production by CD4⁺ T cells was significantly increased （P = 0.003）. Compared with WT mice， the histological score of pancreatic injury， the expression of RhoF and p-MYPT1 in T cells， the number of IL-17⁺ T cells and serum IL-17 level in RhoF transgenic mice were significantly increased （P < 0.05）. The histological score， RhoF and p-MYPT1 expression of T cells， IL-17⁺ T cell numbers and serum IL-17 level were significantly lower in the SAP+Y-27632 group than those in the SAP group （P < 0.05）. Conclusion RhoF/ROCK signaling pathway-mediated polarization of Th17 cells is involved in the pathogenesis of SAP.

Key words: Rho-associated GTP-binding protein F, Th17 cells, severe acute pancreatitis

中图分类号:

R576.023

孙茹雪,朱梦莉,刘晶晶,陈飞. RhoF介导的Th17极化在急性胰腺炎发生中的作用及机制[J]. 实用医学杂志, 2024, 40(10): 1351-1356.

Ruxue SUN,Mengli ZHU,Jingjing LIU,Fei. CHEN. Role and mechanism of RhoF-mediated Th17 polarization in development of acute pancreatitis[J]. The Journal of Practical Medicine, 2024, 40(10): 1351-1356.

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参考文献 22

1	MEDEROS M A， REBER H A， GIRGIS M D. Acute pancreatitis： a review［J］. JAMA， 2021， 325（4）： 382-390. doi:10.1001/jama.2020.20317 doi: 10.1001/jama.2020.20317
2	LEE P J， PAPACHRISTOU G I. New insights into acute pancreatitis［J］. Nat Rev Gastroenterol Hepatol， 2019， 16（8）： 479- 496. doi:10.1038/s41575-019-0158-2 doi: 10.1038/s41575-019-0158-2
3	CHATILA A T， BILAL M， GUTURU P. Evaluation and management of acute pancreatitis［J］. World J Clin Cases， 2019， 7（9）： 1006-1020. doi:10.12998/wjcc.v7.i9.1006 doi: 10.12998/wjcc.v7.i9.1006
4	ZHANG Z， LIU Q， ZANG H， et al. Oxymatrine protects against l-arginine-induced acute pancreatitis and intestine injury involving Th1/Th17 cytokines and MAPK/NF-κB signalling［J］. Pharm Biol， 2019， 57（1）： 595-603. doi:10.1080/13880209.2019.1657906 doi: 10.1080/13880209.2019.1657906
5	KOWLURU A. Roles of GTP and Rho GTPases in pancreatic islet beta cell function and dysfunction［J］. Small GTPases， 2021， 12（5/6）： 323-335. doi:10.1080/21541248.2020.1815508 doi: 10.1080/21541248.2020.1815508
6	SHAVERDASHVILI K， PADLO J， WEINBLATT D， et al. KLF4 activates NFκB signaling and esophageal epithelial inflammation via the Rho-related GTP-binding protein RHOF［J］. PLoS One， 2019， 14（4）： e0215746. doi:10.1371/journal.pone.0215746 doi: 10.1371/journal.pone.0215746
7	LI S， LIU Y， BAI Y， et al. Ras homolog family member F， filopodia associated promotes hepatocellular carcinoma metastasis by altering the metabolic status of cancer cells through RAB3D［J］. Hepatology， 2021， 73（6）： 2361-2379. doi:10.1002/hep.31641 doi: 10.1002/hep.31641
8	SONG J， XI J Y， YU W B， et al. Inhibition of ROCK activity regulates the balance of Th1， Th17 and Treg cells in myasthenia gravis［J］. Clin Immunol， 2019， 203： 142-153. doi:10.1016/j.clim.2019.05.006 doi: 10.1016/j.clim.2019.05.006
9	LV C， HE Y， WEI M， et al. CTRP3 ameliorates cerulein-induced severe acute pancreatitis in mice via SIRT1/NF-κB/p53 axis［J］. Biosci Rep， 2020， 40（10）： BSR20200092. doi:10.1042/bsr20200092 doi: 10.1042/bsr20200092
10	PAN Y， LI Y， GAO L， et al. Development of a novel model of hypertriglyceridemic acute pancreatitis in mice［J］. Sci Rep， 2017， 7（1）： 40799. doi:10.1038/srep40799 doi: 10.1038/srep40799
11	李红英，孟明明. 不同进食时间对急性胰腺炎患者术后疗效及胃肠功能恢复，体液免疫指标的影响［J］. 实用医学杂志， 2021， 37（18）： 2395-2400.
12	MUNIR F， JAMSHED M B， SHAHID N， et al. Advances in immunomodulatory therapy for severe acute pancreatitis［J］. Immunol Lett， 2020， 217： 72-76. doi:10.1016/j.imlet.2019.11.002 doi: 10.1016/j.imlet.2019.11.002
13	STOJANOVIC B， JOVANOVIC I P， STOJANOVIC M D， et al. The Emerging Roles of the Adaptive Immune Response in Acute Pancreatitis［J］. Cells， 2023， 12（11）： 1495. doi:10.3390/cells12111495 doi: 10.3390/cells12111495
14	LI G， CHEN H， LIU L， et al. Role of interleukin-17 in acute pancreatitis［J］. Front Immunol， 2021， 12： 674803. doi:10.3389/fimmu.2021.674803 doi: 10.3389/fimmu.2021.674803
15	GUO J， LI Z， TANG D， et al. Th17/Treg imbalance in patients with severe acute pancreatitis： Attenuated by high-volume hemofiltration treatment［J］. Medicine， 2020， 99（31）： e21491. doi:10.1097/md.0000000000021491 doi: 10.1097/md.0000000000021491
16	DIPANKAR P， KUMAR P， DASH S P， et al. Functional and therapeutic relevance of Rho GTPases in innate immune cell migration and function during inflammation： an in silico perspective［J］. Mediators Inflamm， 2021， 2021： 6655412. doi:10.1155/2021/6655412 doi: 10.1155/2021/6655412
17	HOU Y， ZI J， GE Z. High expression of RhoF predicts worse overall survival： A potential therapeutic target for non-M3 acute myeloid leukemia［J］. J Cancer， 2021， 12（18）： 5530. doi:10.7150/jca.52648 doi: 10.7150/jca.52648
18	LEAL A S， MISEK S A， LISABETH E M， et al. The Rho/MRTF pathway inhibitor CCG-222740 reduces stellate cell activation and modulates immune cell populations in KrasG12D； Pdx1-Cre （KC） mice［J］. Sci Rep， 2019， 9（1）： 7072. doi:10.1038/s41598-019-43430-0 doi: 10.1038/s41598-019-43430-0
19	TAO X， CHEN Q， LI N， et al. Serotonin-RhoA/ROCK axis promotes acinar-to-ductal metaplasia in caerulein-induced chronic pancreatitis［J］. Biomed Pharmacother， 2020， 125： 109999. doi:10.1016/j.biopha.2020.109999 doi: 10.1016/j.biopha.2020.109999
20	LIU P， XIAO Z， YAN H， et al. Baicalin suppresses Th1 and Th17 responses and promotes Treg response to ameliorate sepsis-associated pancreatic injury via the RhoA-ROCK pathway［J］. Int Immunopharmacol， 2020， 86： 106685. doi:10.1016/j.intimp.2020.106685 doi: 10.1016/j.intimp.2020.106685
21	D'ANGELO A， SHIBATA K， TOKUNAGA M， et al. Generation of murine tumour-reactive T cells by co-culturing murine pancreatic cancer organoids and peripheral blood lymphocytes［J］. Biochem Biophys Rep， 2022， 32： 101365. doi:10.1016/j.bbrep.2022.101365 doi: 10.1016/j.bbrep.2022.101365
22	CAI J， ZHOU X， YU H， et al. Effect of bone marrow mesenchymal stem cells on RhoA/ROCK signal pathway in severe acute pancreatitis［J］. Am J Transl Res， 2019， 11（8）： 4809-4816.