胱天蛋白酶募集域蛋白9在重症急性胰腺炎巨噬细胞M1极化中的作用

doi:10.3969/j.issn.1674-8115.2025.08.005

上海交通大学学报（医学版） ›› 2025, Vol. 45 ›› Issue (8): 981-989.doi: 10.3969/j.issn.1674-8115.2025.08.005

胱天蛋白酶募集域蛋白9在重症急性胰腺炎巨噬细胞M1极化中的作用

王琳¹, 徐萍¹^,², 张乔婷², 田军¹, 娄晓丽², 王静¹()

^1.上海交通大学医学院附属松江医院消化内科，上海 201600
^2.南京医科大学上海松江临床医学院，上海 201600

收稿日期:2024-12-06 接受日期:2025-04-25 出版日期:2025-08-28 发布日期:2025-08-18
通讯作者: 王静，主任医师，博士，电子信箱: wangj0081@126.com。
基金资助:
上海市自然科学基金(20ZR1450900)

Role of CARD9 in macrophage M1 polarization in severe acute pancreatitis rats

WANG Lin¹, XU Ping¹^,², ZHANG Qiaoting², TIAN Jun¹, LOU Xiaoli², WANG Jing¹()

^1.Department of Gastroenterology, Songjiang Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201600, China
^2.Shanghai Songjiang Clinical Medical College of Nanjing Medical University, Shanghai 201600, China

Received:2024-12-06 Accepted:2025-04-25 Online:2025-08-28 Published:2025-08-18
Contact: Wang Jing, E-mail: wangj0081@126.com.
Supported by:
Shanghai Natural Science Foundation(20ZR1450900)

摘要/Abstract

摘要：

目的·探讨胱天蛋白酶募集域蛋白9（caspase recruitment domain-containing protein 9，CARD9）是否参与调控重症急性胰腺炎（severe acute pancreatitis，SAP）巨噬细胞极化。方法·将SD大鼠随机分为4组，分别为对照组、SAP组、SAP+CARD9 shRNA组和SAP+Control shRNA组，每组6只。在SAP大鼠造模前48 h，经尾静脉注射CARD9 shRNA干扰腺病毒。造模后12 h收集标本，采用H-E染色评估胰腺组织病理变化，运用real-time PCR法检测大鼠腹腔巨噬细胞CARD9及肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-10、精氨酸酶-1（arginase 1，Arg-1）的基因表达水平，通过Western blotting法检测腹腔巨噬细胞CARD9蛋白表达水平，利用流式细胞技术检测大鼠腹腔巨噬细胞极化类型。结果·SAP组、SAP+CARD9 shRNA组、SAP+Control shRNA组中CARD9基因及CARD9蛋白表达水平均明显高于对照组，显示CARD9干扰成功（P<0.001）。SAP+CARD9 shRNA组与SAP组相比：血清淀粉酶水平、胰腺组织病理评分明显降低；腹腔巨噬细胞促炎细胞因子TNF-α、IL-6水平明显降低，抗炎细胞因子IL-10、Arg-1水平升高；M1型巨噬细胞明显减少、M1/M2比值降低。腹腔巨噬细胞CARD9 mRNA水平与TNF-α mRNA、IL-6 mRNA水平及M1型巨噬细胞数量呈正相关。结论·CARD9促进SAP大鼠巨噬细胞M1型极化；下调CARD9表达可通过抑制巨噬细胞M1极化，减轻SAP大鼠炎症反应。

关键词: 巨噬细胞极化, 重症急性胰腺炎, 胱天蛋白酶募集域蛋白9, SD大鼠, 腹腔巨噬细胞

Abstract:

Objective ·To investigate the role of caspase recruitment domain-containing protein 9 (CARD9) in regulating macrophage polarization in a rat model of severe acute pancreatitis (SAP). Methods ·SD rats were divided into 4 groups: Control group, SAP group, SAP+CARD9 shRNA group, and SAP+Control shRNA group with six rats in each group. The SAP rats transfected with CARD9 shRNA were established by injecting CARD9 shRNA adenovirus 48 hours before the SAP model was induced. The pancreatic tissues, peripheral blood, and peritoneal macrophages were collected 12 hours after the model was established. The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages and pancreatic tissues were measured by real-time PCR and Western blotting. The expressions of TNF-α, IL-6, IL-10 and Arg-1 mRNA were detected by real-time PCR and the polarization types of peritoneal macrophages were detected by flow cytometry. Results ·The expressions of CARD9 gene and CARD9 protein in peritoneal macrophages in CARD9 shRNA rats were significantly lower than those in SAP rats and interference control rats, which confirmed the success of CARD9 interference model. Compared with SAP rats, CARD9 shRNA rats had significantly reduced degree of inflammation and pathological scores; the mRNA levels of TNF-α, and IL-6 in peritoneal macrophages were significantly decreased; meanwhile, the mRNA levels of IL-10 and Arg-1 were increased, and the changes in TNF-α and IL-6 were significantly higher than those of IL-10 and Arg-1. The proportion of M1 macrophages was significantly reduced, and the ratio of M1/M2 was significantly decreased. The expression level of CARD9 mRNA in peritoneal macrophages was positively correlated with the proportion of M1 macrophages and the mRNA levels of TNF-α and IL-6. Conclusion ·CARD9 is involved in regulating macrophage polarization in SAP rats, and it mainly regulates M1 polarization. Inhibition of CARD9 expression can reduce M1 macrophage polarization and reduce the inflammatory response in SAP rats.

Key words: macrophage polarization, severe acute pancreatitis, caspase recruitment domain-containing protein 9, SD rat, peritoneal macrophage

中图分类号:

R576

王琳, 徐萍, 张乔婷, 田军, 娄晓丽, 王静. 胱天蛋白酶募集域蛋白9在重症急性胰腺炎巨噬细胞M1极化中的作用[J]. 上海交通大学学报（医学版）, 2025, 45(8): 981-989.

WANG Lin, XU Ping, ZHANG Qiaoting, TIAN Jun, LOU Xiaoli, WANG Jing. Role of CARD9 in macrophage M1 polarization in severe acute pancreatitis rats[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(8): 981-989.

图/表 6

图1 4组大鼠腹腔巨噬细胞 CARD9 mRNA及CARD9蛋白表达变化Note： A. Expression of CARD9 protein in peritoneal macrophages of the four groups of rats. B. Expression of CARD9 mRNA in peritoneal macrophages of the four groups of rats. C. CARD9 protein expression. ①P<0.001; ②P<0.001, compared with the control group.

Fig1 Expression of CARD9 mRNA and CARD9 protein in peritoneal macrophages of the four groups of rats

图2 4组大鼠血清淀粉酶水平比较Note： ①P<0.001; ②P<0.001, compared with the control group.

Fig 2 Comparison of serum amylase levels among the four groups of rats

图3 4组大鼠胰腺组织病理改变及病理评分（H-E染色, ×200）Note： A. Pathological changes in pancreatic tissue (the area indicated by the arrow shows regions with more severe structural necrosis). B. Pathology score of the pancreas. ①P<0.001; ②P<0.001, compared with the control group.

Fig 3 Pathological changes and scores of pancreatic tissues in the four groups of rats（H-E staining, ×200）

图4 4组大鼠腹腔巨噬细胞细胞因子转录水平变化Note： A. Changes in the transcriptional levels of TNF-α in the four groups of rats. B. Changes in the transcriptional levels of IL-10in the four groups of rats. C. Changes in the transcriptional levels of IL-6 in the four groups of rats. D. Changes in the transcriptional levels of Arg-1in the four groups of rats. ①P<0.001; ②P<0.001, compared with the control group; ③P=0.004, compared with the SAP group; ④P =0.014, compared with the control group.

Fig 4 Changes in the transcriptional levels of cytokines in peritoneal macrophages of the four groups of rats

图5 4组大鼠腹腔巨噬细胞极化状态Note： A. Flow cytometry scatter distribution of CCR2 expression in the four groups (CCR2 represents M1 macrophage polarization). B. Flow cytometry scatter distribution of CD206 in four groups (CD206 represents M2 macrophage polarization). C. Changes in the polarization status of macrophages in the four groups. ①P<0.001; ②P<0.001, compared with the control group.

Fig 5 Polarization of peritoneal macrophages in the four groups of rats

图6 腹腔巨噬细胞极化状态与 CARD9 的相关性Note： A. The correlation between TNF-α and CARD9. B. The correlation between IL-6and CARD9. C. The correlation between IL-10 and CARD9. D, The correlation between Arg-1 and CARD9.

Fig 6 Correlation between the polarization state of peritoneal macrophages and CARD9

参考文献 5

[1]	中华医学会急诊医学分会, 上海市医学会急诊专科分会. 急性胰腺炎急诊诊治专家共识[J]. 中华急诊医学杂志, 2024, 33(4): 470-479.
	Emergency Medicine Branch of the Chinese Medical Association, Emergency Medicine Branch of Shanghai Medical Association. Expert consensus on the emergency diagnosis and treatment of acute pancreatitis[J].Chinese Journal of Emergency Medicine, 2024, 33(4): 470-479.
[2]	ZHAO Q L, WEI Y, PANDOL S J, et al. STING signaling promotes inflammation in experimental acute pancreatitis[J]. Gastroenterology, 2018, 154(6): 1822-1835.e2.
[3]	YANG Z W, WENG C Z, WANG J, et al. The role of CARD9 overexpression in peripheral blood mononuclear cells from patients with aseptic acute pancreatitis[J]. J Cell Mol Med, 2016, 20(3): 441-449.
[4]	王琳, 徐萍, 王静, 等. 同种异体骨髓间充质干细胞对急性坏死性胰腺炎大鼠腹腔巨噬细胞极化的影响[J]. 中华胰腺病杂志, 2020(2): 120-125.
	WANG L,XU P,WANG J,et al.Effect of allogeneic bone marroww mesenchymal stem cells on macrophage polarization in rats with acute necrosis pancreatitis[J].Chinese Journal of Pancreatic Disease, 2020, 20(2): 120-125.
[5]	YANG M, SHAO J H, MIAO Y J, et al. Tumor cell-activated CARD9 signaling contributes to metastasis-associated macrophage polarization[J]. Cell Death Differ, 2014, 21(8): 1290-1302.

[1]	牛媛媛, 汪龙德, 胥文娟, 李正菊, 张瑞婷, 吴毓谦. 巨噬细胞M1/M2型极化在不同肝病中的作用研究进展[J]. 上海交通大学学报（医学版）, 2024, 44(4): 509-517.
[2]	李旭冉, 陶诗聪, 郭尚春. 骨髓间充质干细胞来源小细胞外囊泡对骨质疏松症的改善作用[J]. 上海交通大学学报（医学版）, 2023, 43(4): 406-416.
[3]	孔祥歆,刘卉芳,陈凤玲 . CaMKKβ 通过激活 AMPK/JAK2/STAT3 信号促进小鼠单核巨噬细胞向 M2 表型转换[J]. 上海交通大学学报（医学版）, 2017, 37(7): 914-.
[4]	刘晓颖, 黄洁, 费健, 等. 重症急性胰腺炎死亡患者的临床特征分析[J]. , 2013, 33(5): 640-.
[5]	刘晓颖, 陈尔真. 新日本严重度评分评估重症急性胰腺炎严重度及风险的价值[J]. , 2012, 32(3): 344-.
[6]	黄洁, 毛恩强. 暴发性急性胰腺炎早期感染特点分析[J]. , 2010, 30(10): 1267-.
[7]	娄晓丽, 张彦, 何平, 等. 豚鼠腹腔巨噬细胞吞噬钩端螺旋体的机制研究[J]. , 2009, 29(10): 1143-.

胱天蛋白酶募集域蛋白9在重症急性胰腺炎巨噬细胞M1极化中的作用

Role of CARD9 in macrophage M1 polarization in severe acute pancreatitis rats

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