黑色素瘤缺乏因子2介导的细胞焦亡通路在特发性炎性肌病患者外周血单个核细胞中的表达

doi:10.19723/j.issn.1671-167X.2026.01.023

摘要/Abstract

摘要：

目的: 检测特发性炎性肌病(idiopathic inflammatory myopathy，IIM)患者外周血单个核细胞(peripheral blood mononuclear cell，PBMC)中黑色素瘤缺失因子2(absent in melanoma 2，AIM2)及其介导的细胞焦亡通路关键组分——半胱氨酸天冬氨酸蛋白酶1(cysteine aspartate-specific protease-1，caspase-1)和焦孔素蛋白D(gasdermin D，GSDMD)的表达，并探讨其在IIM发病机制中的作用。方法: 招募2020年5月至2022年6月于中国人民解放军北部战区总医院风湿免疫科就诊的30例IIM患者(IIM组)，同期于医院体检中心招募30名性别和年龄与IIM患者相匹配的健康志愿者(健康对照组)，收集研究对象临床信息、血液生化和免疫标志物，以及静脉血样本。通过荧光定量法检测血清双链DNA(double-stranded DNA，dsDNA)水平，通过实时荧光定量逆转录PCR(reverse transcription quantitative real-time PCR，RT-qPCR)检测PBMC中AIM2、caspase-1、GSDMD、白细胞介素1β(interleukin 1β，IL-1β)和IL-18的mRNA表达水平，应用蛋白免疫印迹法检测PBMC中AIM2、caspase-1、GSDMD、IL-1β和IL-18的蛋白表达水平，应用酶联免疫吸附(enzyme-linked immuno-sorbent assay，ELISA)法检测血清中IL-1β和IL-18的表达水平。结果: IIM组包含皮肌炎(dermatomyositis，DM，n=10)、多发性肌炎(polymyositis，PM，n=5)、重叠性肌炎(overlap syndrome，OM，n=11)和免疫介导的坏死性肌病(immune-mediated necrotizing myopathy，IMNM，n=4)4个亚组。与健康对照组相比，IIM组及其各亚组的血清中dsDNA、IL-1β和IL-18水平均显著增加(P＜0.05)。除IMNM亚组PBMC中AIM2 mRNA与健康对照组差异无统计学意义外，IIM组及其他亚组AIM2、caspase-1和GSDMD mRNA表达均显著增加(P＜0.05)；除IMNM和OM亚组PBMC中IL-1β mRNA与健康对照组比较差异无统计学意义外，IIM组及其他亚组IL-1β和IL-18 mRNA表达均显著增加(P＜0.05)；亚组间比较表明，DM亚组PBMC中IL-1β mRNA表达明显高于OM和IMNM亚组，PM亚组IL-18 mRNA表达明显高于DM和OM亚组(P＜0.05)。IIM及其各亚组的PBMC中，AIM2、caspase-1、GSDMD、IL-1β和IL-18蛋白的表达水平均显著高于健康对照组(P＜0.05)；各亚组间比较发现，OM亚组的IL-18蛋白表达显著高于PM亚组(P＜0.05)。相关性分析表明，IIM组caspase-1、GSDMD和IL-18 mRNA与AIM2 mRNA呈正相关，caspase-1、GSDMD、IL-1β和IL-18蛋白与AIM2蛋白表达也呈正相关。结论: AIM2炎性小体介导的细胞焦亡通路可能参与IIM的发病机制，这一结论可以为研究IIM的病因及开发新的治疗方法提供理论基础。

关键词: 特发性炎性肌病, 黑色素瘤缺乏因子2, 细胞焦亡, 半胱氨酸天冬氨酸蛋白酶1, 焦孔素蛋白D

Abstract:

Objective: To detect the expression levels of absence in melanoma 2 (AIM2), cysteine aspartate-specific protease-1 (caspase-1), and gasdermin D (GSDMD) in peripheral blood mononuclear cell (PBMC) of patients with idiopathic inflammatory myopathy (IIM) and to explore their role in the pathogenesis of IIM. Methods: A total of 30 IIM patients (IIM group) who visited the Department of Rheumatology and Immunology, General Hospital of Northern Theater Command from May 2020 to June 2022 were recruited. Concurrently, 30 healthy volunteers matched by gender and age were recruited from the hospital's Health Examination Center. Clinical information, biochemical and immunological mar-kers, and venous blood samples were collected from the study subjects. Serum double-stranded DNA (dsDNA) levels were detected by fluorescence quantitative method, and the mRNA expression levels of AIM2, caspase-1, GSDMD, interleukin 1β (IL-1β), and IL-18 in PBMC were detected by reverse transcription quantitative real-time PCR (RT-qPCR). The protein expression levels of AIM2, caspase-1, GSDMD, IL-1β, and IL-18 in PBMC were detected using the Western blot (WB) method, and the serum levels of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay (ELISA). Results: The IIM group included 10 cases of dermatomyositis (DM), 5 cases of polymyositis (PM), 11 cases of overlap syndrome (OM), and 4 cases of immune-mediated necrotizing myopathy (IMNM). Compared with the healthy control group, the serum levels of dsDNA, IL-1β, and IL-18 were significantly increased in the IIM group and its subgroups (P < 0.05). Except for the fact that there was no statistically significant difference in AIM2 mRNA levels in PBMC of the IMNM subgroup compared to the healthy control group, the expression of AIM2, caspase-1, and GSDMD mRNA was significantly increased in the IIM group and other subgroups (P < 0.05); Except for the comparison of IL-1β mRNA levels in PBMC of the IMNM and OM subgroups with the healthy control group showing no statistical difference, the expression of IL-1β and IL-18 mRNA was significantly increased in the IIM group and other subgroups (P < 0.05); Comparisons between subgroups indicated that the expression of IL-1β mRNA in the DM subgroup was significantly higher than that in the OM and IMNM subgroups, and the expression of IL-18 mRNA in the PM subgroup was significantly higher than that in the DM and OM subgroups (P < 0.05). The expression levels of AIM2, caspase-1, GSDMD, IL-1β, and IL-18 proteins in PBMC of the IIM group and its subgroups were significantly higher than those in the healthy control group (P < 0.05); Comparisons among subgroups revealed that the expression of IL-18 protein in the OM subgroup was significantly higher than that in the PM subgroup (P < 0.05). In the IIM group, the mRNA of caspase-1, GSDMD, and IL-18 showed a positive correlation with AIM2 mRNA, and the protein expression of caspase-1, GSDMD, IL-1β, and IL-18 also showed a positive correlation with AIM2 protein expression. Conclusion: The AIM2 inflammasome-mediated pyroptosis pathway may be involved in the pathogenesis of IIM, providing a theoretical basis for further research on the etiology of IIM and the development of new therapies.

Key words: Idiopathic inflammatory myopathy, Absent in melanoma 2, Pyroptosis, Caspase-1, Gasdermin D

中图分类号:

R593.26

初吉燕, 李萍, 田竞, 付笛语, 郭琳, 孙蕊, 李亚娣. 黑色素瘤缺乏因子2介导的细胞焦亡通路在特发性炎性肌病患者外周血单个核细胞中的表达[J]. 北京大学学报（医学版）, 2026, 58(1): 175-183.

Jiyan CHU, Ping LI, Jing TIAN, Diyu FU, Lin GUO, Rui SUN, Yadi LI. Expression of the melanoma 2-mediated pyroptosis pathway in peripheral blood mononuclear cells of patients with idiopathic inflammatory myopathies[J]. Journal of Peking University (Health Sciences), 2026, 58(1): 175-183.

图/表 6

表1

表2

健康对照组和IIM组及其各亚组患者临床资料与实验室指标比较"

Items	IIM group
Items	DM subgroup (n = 10)	PM subgroup (n = 5)	OM subgroup (n = 11)	IMNM subgroup (n = 4)	Total (n = 30)	HC group (n = 30)
Demographic characteristics
Female	9 (90.0)	3 (60.0)	8 (72.7)	2 (50.0)	22 (73.3)	20 (66.7)
Age/years	59.3 ± 11.7	52.6 ± 3.9	55.3 ± 15.5	65.5 ± 13.5	55.0 ± 15.1	53.1 ± 8.4
Duration/months	5.5 (3.5 – 12.3)	24.0 (8.5 – 66.0)	12.0 (1.0 – 180.0)	14.0 (5.0 – 14.8)
Assessment of disease severity and activity
VAS score	76.8 ± 9.4	75.7 ± 5.1	69.7 ± 21.5	68.3 ± 38.8
MMT score	65.1 ± 14.6	75.0 ± 3.0	66.0 ± 17.4	68.7 ± 16.3
MYOACT score	3.2 ± 0.9	2.6 ± 1.2	4.4 ± 1.2	3.9 ± 0.7
MDAAT score	7.0 (3.8 – 8.3)	6.0 (4.3 – 8.5)	6.0 (4.5 – 8.0)	4.0 (3.3 – 6.3)
MDI score	3.0 (2.0 – 7.5)	5.0 (3.0 – 7.5)	9.0 (4.0 – 19.0)	2.5 (1.3 – 6.0)
Laboratory indicators
WBC/(×10⁹/L)	6.6 (6.0 – 7.4)	7.0 (4.7 – 11.3)	10.1 (5.5 – 12.4)	10.1 (5.5 – 12.4)	6.9 (6.1 – 8.7)^*	5.9 (5.0 – 6.5)
LY/(×10⁹/L)	1.1 (0.5 – 28.2)^^	0.8 (0.7 – 2.1)	1.5 (0.7 – 1.8)	1.6 (1.0 – 1.7)	1.2 (0.5 – 28.2)	1.4 (0.3 – 3.5)
Hb/(g/L)	119.3 ± 17.1^*	136.2 ± 15.3	126.9 ± 18.4	130.3 ± 9.4	126.4 ± 16.9^*	140.0 ± 12.7
ALT/(U/L)	50.3 (9.0 – 290.4)^^	123.0 (47.0 – 340.0)^*	31.0 (11.0 – 340.0)	81.0 (80.0 – 410.9)^^^▲	55.5 (9.0 – 410.9)^*	25.5 (10.0 – 89.0)
AST/(U/L)	104.0 (12.0 – 441.7)^^	107.0 (27.0 – 266.0)^*	25.0 (10.0 – 136.0)△	106.0 (73.0 – 589.1)^^^▲	76.0 (10.0 – 441.7)^*	20.5 (13.0 – 55.0)
LDH/(U/L)	513.5 (169.0 – 808.0)^^	543.0 (184.0 – 1335.0)^^	255.0 (162.0 – 1595.0)^^	1858.5 (891.0 – 1957.0)^^^▲^▲	551.0 (162.0 – 1957.0)^*	174.0 (98.0 – 278.0)
CK/(U/L)	442.0 (27.0 – 716.2)^^	2 554.0 (55.0 – 7690.0)^^	149.0 (36.0 – 1031.0)^^△	3428.5 (131.0 – 1679.0)^^^▲^▲	488.0 (27.0 – 1639.0)^*	90.0 (21.0 – 178.0)
CK-MB/(U/L)	54.0 (13.1 – 573.5)^^	60.8 (9.8 – 432.3)^^	12.1 (9.0 – 54.1)^^△	200.0 (181.4 – 899.2)^^^▲^▲	31.35 (9.0 – 899.0)^*	12.0 (3.0 – 23.4)
TNT/(μg/L)	27.0 (5.0 – 650.0)^^	200.0 (5.0 – 2820.0)^^	61.0 (5.0 – 279.0)^^	611.0 (383.0 – 2700.0)^^^▲^▲	92.0 (5.0 – 650.0)^*	0.0 (0.0 – 10.0)
ESR/(mm/h)	19.0 (2.0 – 58.0)^^	15.0 (2.0 – 35.0)^^	11.0 (2.0 – 106.0)^^	13.0 (10.0 – 46.0)^^	13.5 (2.0 – 106.0)^*	2.0 (2.0 – 12.0)
CRP/(mg/L)	4.9 (2.9 – 59.3)^^	2.9 (1.3 – 11.0)	4.4 (1.3 – 81.5)^^	9.1 (0.6 – 9.2)	4.0 (0.6 – 81.5)^*	0.9 (0.2 – 2.5)
SF/(mg/L)	187.5 (30.6 – 578.2)^^	255.5 (30.62 – 1632.0)^^	463.1 (142.0 – 1433.0)^^	626.3 (503.7 – 1345.0)^^^▲^▲	364.8 (30.6 – 1632.0)^*	67.4 (15.8 – 176.5)
IgG/(g/L)	16.9 ± 4.4^*	11.1 ± 4.6^*	11.6 ± 4.9^*	14.0 ± 0.6	13.6 ± 4.9^*	11.3 ± 3.6
IgA/(g/L)	3.3 (2.0 – 5.0)^^	1.8 (1.0 – 2.6)^^	2.2 (0.9 – 3.4)	2.3 (1.9 – 3.2)	2.3 (0.6 – 6.6)	2.0 (0.8 – 4.1)
IgM/(g/L)	1.3 (0.8 – 1.7)	1.1 (0.3 – 2.3)	1.0 (0.7 – 1.3)	1.5 (1.2 – 1.8)^^^▲^▲	1.1 (0.8 – 1.5)	1.0 (0.8 – 1.4)
ANA positive	6 (60)	4 (80)	9 (82)	2 (50)	21 (70)	0

表2

图1

图2

图3

图4

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Gene	Forward primer	Reverse primer
GAPDH	5′-ACAACTTTGGTATCGTGGAAGG-3′	5′-GCCATCACGCCACAGTTTC-3′
AIM2	5′-AGCAAGATATTATCGGCACAGTG-3′	5′-GTTCAGCGGGACATTAACCTT-3′
caspase-1	5′-TTTCCGCAAGGTTCGATTTTCA-3′	5′-GGCATCTGCGCTCTACCATC-3′
GSDMD	5′-GTGTGTCAACCTGTCTATCAAGG-3′	5′-CATGGCATCGTAGAAGTGGAAG-3′
IL-1β	5′-TGAGCTCGCCAGTGAAATGAT-3′	5′-TGCTGTAGTGGTGGTCGGAG-3′
IL-18	5′-TTCAAGACCAGCCTGACCAAC-3′	5′-GCTCACCACAACCTCTACCTCC-3′

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