银屑病中滤泡性辅助性T细胞及IL-21对HaCaT细胞增殖及细胞周期的影响

doi:10.3969/j.issn.1006-5725.2023.18.010

摘要/Abstract

摘要：

目的探究寻常型银屑病患者外周血滤泡性辅助性T细胞及其主要功能性细胞因子IL-21在体外培养条件下对HaCaT细胞增殖及细胞周期的影响。方法选取15例寻常型银屑病患者作为试验组，同时纳入10位健康志愿者作为对照组。运用流式细胞技术分选出外周血CD4⁺CXCR5⁺的Tfh细胞，与HaCaT细胞以1∶1进行共培养。加入IL-21或IL-21中和抗体干预72 h后收集HaCaT细胞。采用CCK-8法检测细胞增殖活性，并运用流式细胞学技术检测HaCaT细胞周期及Ki-67表达特征。结果 IL-21对HaCaT细胞的增殖有促进作用（P < 0.05），使G₀/G₁期HaCaT细胞减少、S期HaCaT细胞增多（P < 0.05）。HaCaT细胞与Tfh细胞共培养时，银屑病组与健康对照组HaCaT细胞增殖活性均较空白对照组显著升高（P < 0.000 1），且银屑病组较健康对照组促进作用更强（P < 0.05）；银屑病组与健康对照组中加入IL-21组均比加入IL-21中和抗体组的HaCaT细胞增殖活性高（P < 0.05）；此外，在银屑病组细胞共培养中，IL-21中和抗体组同单纯共培养组相比，可将HaCaT细胞阻滞于G₀/G₁期，减少S期细胞数量，进而抑制HaCaT 细胞增殖活性（P < 0.05）。结论银屑病患者中Tfh细胞及其主要功能性细胞因子IL-21可促进角质形成细胞增殖，且中和IL-21会抑制这种促进作用。因此，Tfh细胞可能通过IL-21促进角质形成细胞增殖从而促进银屑病的发生发展。

关键词: 银屑病, 滤泡性辅助性T细胞, HaCaT细胞, 白细胞介素-21

Abstract:

Objective To observe the effects of T follicular helper cells and the main functional cytokines IL?21 on the proliferation and cell cycle of HaCaT cells cultured in vitro in patients with psoriasis. Methods Fifteen patients with psoriasis vulgaris and 10 healthy controls were defined as psoriasis group and healthy control group， respectively. CD4⁺CXCR5⁺ Tfh cells from psoriatic patients and healthy controls were separated by flow cytometry and co?cultured with HaCaT cells at 1∶1. IL?21 or IL?21 neutralizing antibodies were added and the HaCaT cells were collected after 72 h of intervention. The cell proliferation was measured by the CCK?8 and the cell cycle and Ki?67 expression of HaCaT cells were detected by flow cytometry. Results IL?21 promoted the proliferation of HaCaT cells （P < 0.05） and increased the proportion of HaCaT cells in S phase and decreased the proportion of cells in G₀/G₁ phase （P < 0.05）. When Tfh cells were co?cultured with HaCaT cells， the proliferation activity of HaCaT cells in psoriasis group and healthy control group was significantly higher than that in control group （P < 0.000 1）， and the proliferation activity of HaCaT cells in psoriasis group was higher than that in IL?21 neutralization antibody group （P < 0.000 1）. In psoriasis group， IL?21 neutralizing antibody group compared with Tfh?HaCaT co?culture group show that it could block HaCaT cells in G₀/G₁ phase， reduce the number of cells in S phase， and then inhibit the proliferation of HaCaT cells. Conclusion IL?21 promoted the HaCaT cell proliferation invitro； Tfh cells inpsoriasispatients promoted the proliferation of keratinocytes invitro， and blocking IL?21 inhibitd this promoting effect. In summary， Tfh cells may promote theproliferation of keratinocytes through producing IL?21， thereby promoting the development and progression of psoriasis.

Key words: psoriasis, T follicular helper cells, HaCaT cells, interleukin?21

中图分类号:

R758.63

李思彤,王傲,白彦萍,王英. 银屑病中滤泡性辅助性T细胞及IL-21对HaCaT细胞增殖及细胞周期的影响[J]. 实用医学杂志, 2023, 39(18): 2342-2348.

Sitong LI,Ao WANG,Yanping BAI,Ying. WANG. Effects of T follicular helper cells and IL⁃21 on proliferation and cell cycle of HaCaT cellsinpsoriasis[J]. The Journal of Practical Medicine, 2023, 39(18): 2342-2348.

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分组	细胞周期			PI
分组	G₀/G₁	S	G₂/M	PI
P值	< 0.05	< 0.05	> 0.05	< 0.05
HaCaT	57.81 ± 10.51	32.85 ± 8.56	9.34 ± 3.71	42.19 ± 10.51
IL-21+HaCaT	47.01 ± 9.02	43.73 ± 8.37	9.26 ± 2.90	52.99 ± 9.01

分组	细胞增殖活性（od450）		Ki-67（%）
分组	HC	PV	HC	PV
Tfh+HaCaT	2.49 ± 0.94	3.42 ± 0.94	53.80 ± 22.48	64.29 ± 10.45
IL-21	3.04 ± 0.73	3.57 ± 0.69	68.61 ± 17.78	61.28 ± 22.01
IL-21中和抗体	2.46 ± 0.58	2.47 ± 0.96	48.45 ± 20.61	42.13 ± 18.92

分组	细胞周期（%）			PI
分组	G₀/G₁	S	G₂/M	PI
HC-T+H	54.33 ± 10.18	37.11 ± 10.79	8.57 ± 3.86	45.68 ± 10.18
HC-T+H+I	47.13 ± 9.86	41.77 ± 7.62	11.10 ± 3.13	52.87 ± 9.86
HC-T+H+A	58.98 ± 9.62	33.06 ± 9.77	7.95 ± 2.58	41.01 ± 9.62
PV-T+H	46.18 ± 6.03	43.86 ± 9.00	9.96 ± 4.48	53.82 ± 6.03
PV-T+H+I	49.96 ± 6.46	39.50 ± 6.91	10.55 ± 3.43	50.04 ± 6.46
PV-T+H+A	56.93 ± 8.91	33.54 ± 8.19	9.53 ± 2.31	43.07 ± 8.91