寻常型银屑病患者血清中IL-17的表达及紫草素对IL-17刺激HaCaT细胞分泌IL-6和IL-23的影响

doi:10.13481/j.1671-587x.20170519

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (05): 958-962.doi: 10.13481/j.1671-587x.20170519

寻常型银屑病患者血清中IL-17的表达及紫草素对IL-17刺激HaCaT细胞分泌IL-6和IL-23的影响

行敏^1,2, 任宏伟^1,3, 屈慧明^1,4, 耿龙¹

1. 中国医科大学附属第一医院皮肤性病科, 辽宁沈阳 110001;
2. 抚顺矿务局总医院皮肤性病科, 辽宁抚顺 113008;
3. 抚顺矿务局总医院儿科, 辽宁抚顺 113008;
4. 第四军医大学唐都医院皮肤性病科, 陕西西安 710038

收稿日期:2017-01-19 出版日期:2017-09-28 发布日期:2017-09-29
通讯作者: 耿龙,教授,博士研究生导师(Tel:024-83282196,E-mail:genglong99@126.com) E-mail:genglong99@126.com
作者简介:行敏(1981-),女,山西省侯马市人,主治医师,医学硕士,主要从事皮肤病基础和临床方面的研究。
基金资助:
国家自然科学基金资助课题（81673055）；辽宁省高等学校优秀人才支持计划项目资助课题（LR2012026）

Clinical significance of IL-17 expression in serum of patients with psoriasis vulgaris and effects of shikonin on IL-6 and IL-23 production in IL-17-stimulated HaCaT cells

HANG Min^1,2, REN Hongwei^1,3, QU Huiming^1,4, GENG Long¹

1. Department of Dermatology and Venereology, First Affiliated Hospital, China Medical University, Shenyang 110001, China;
2. Department of Dermatology and Venereology, General Hospital, Fushun Mining Bureau, Fushun 113008, China;
3. Department of Paediatrics and Venereology, General Hospital, Fushun Mining Bureau, Fushun 113008, China;
4. Department of Dermatology and Venereology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China

Received:2017-01-19 Online:2017-09-28 Published:2017-09-29

摘要/Abstract

摘要： 目的：探讨寻常型银屑病患者血清中白细胞介素17（IL-17）的表达水平及IL-17刺激后HaCaT细胞分泌白细胞介素6（IL-6）和白细胞介素23（IL-23）的变化，阐明其临床意义及紫草素的干预作用。方法：以25名正常对照者、29例寻常型银屑病患者和不同组别HaCaT细胞（空白对照组、IL-17刺激24h组、IL-17刺激36h组、IL-17刺激48h组、紫草素+IL-17组、环孢素A+IL-17组和IL-17组）为研究对象，采用双抗体夹心酶联免疫吸附法（ELISA）检测寻常型银屑病患者血清IL-17水平和HaCaT细胞各组上清液中IL-6、IL-23水平；实时荧光免疫定量-聚合酶链式反应法（RT-PCR）检测各组HaCaT细胞中IL-6和IL-23p19 mRNA表达水平；采用细胞计数盒-8（CCK-8）法检测各组HaCaT细胞活力。结果：与正常对照组比较，银屑病组患者血清IL-17水平明显升高，以重度皮损银屑病组升高最为明显（P < 0.05）；IL-17刺激24、36和48h组HaCaT细胞及其培养上清中IL-6和IL-23水平及其mRNA表达水平均高于空白对照组（P < 0.01）；紫草素+IL-17组和环孢素A+IL-17组HaCaT细胞及其培养上清中IL-6和IL-23水平及其mRAN表达水平均低于IL-17组（P < 0.05）。与空白对照组比较，其他各组细胞活力差异无统计学意义（P > 0.05）。结论：银屑病患者血清IL-17表达水平升高，尤其是重度皮损银屑病患者血清IL-17表达水平升高明显，IL-17可促进HaCaT细胞分泌IL-6和IL-23，呈时间依赖性，紫草素可抑制IL-17的促炎症作用。

关键词: 银屑病, HaCaT细胞, 白细胞介素17, 白细胞介素6, 白细胞介素23, 紫草素

Abstract: Objective: To explore the expression level of serum interleukin-17 (IL-17) in the patients with psoriasis vulgaris and the changes of interleukin-6 (IL-6) and interleukin-23 (IL-23) levels secrered by the HaCaT cells after stimulated with IL-17, and to clarify its clinical significance and the intervention effect of shikonin. Methods: Twenty-five normal controls, 29 patients with psoriasis vulgaris and different groups of HaCaT cells (blank control group,IL-17-24 h group,IL-17-36 h group,IL-17-48 h group,shikonin+IL-17 group,CsA+IL-17 group and IL-17 group) were used as the subjects. The levels of serum IL-17 in the patients with psoriasis vulgaris and the levels of IL-6 and IL-23 in supernatant of HaCaT cells in various groups were measured by double antibody sandwich enzyme-linked immunosorbent assay(ELISA) and real-time polymerase chain reaction RT-PCR was used to detect the expression levels of IL-6 and IL-23 p19 mRNA in HaCaT cells in various groups.At the same time,Cell Counting Kit-8 (CCK-8) method was used to detect the viabilities of HaCaT cells in various groups. Results: The level of serum IL-17 of the patients in psoriasis vulgaris group was increased compared with nomral control group, especially in the severe skin lesion group(P<0.05). The levels of IL-6 and IL-23 in HaCaT cells and supernatant and their mRNA expression levels in IL-17-24 h,IL-17-36 h and IL-17-48 h groups were significantly higher than those in blank control group (P<0.01). The expression levels of IL-6 and IL-23 in HaCaT cells and supernatant and their mRNA expression levels in shikonin+IL-17 and CsA+IL-17 groups were lower than those in IL-17 groups (P<0.05). No statistical differences were found in the cell viabilities between each drug treatment group and blank control group (P>0.05). Conclusion: The expression level of IL-17 in the patients with psoriasis vulgaris is significantly increased,especially in the patients with severe psoriasis vulgaris.IL-17 can promote the secretion of IL-6 and IL-23 in HaCaT cells in a time-dependent manner. Shikonin can inhibit the proinflammatory effect of IL-17.

Key words: psoriasis, HaCaT cell, interleukin-17, interleukin-6, interleukin-23, shikonin

中图分类号:

R758.63

行敏, 任宏伟, 屈慧明, 耿龙. 寻常型银屑病患者血清中IL-17的表达及紫草素对IL-17刺激HaCaT细胞分泌IL-6和IL-23的影响[J]. 吉林大学学报(医学版), 2017, 43(05): 958-962.

HANG Min, REN Hongwei, QU Huiming, GENG Long. Clinical significance of IL-17 expression in serum of patients with psoriasis vulgaris and effects of shikonin on IL-6 and IL-23 production in IL-17-stimulated HaCaT cells[J]. Journal of Jilin University Medicine Edition, 2017, 43(05): 958-962.

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寻常型银屑病患者血清中IL-17的表达及紫草素对IL-17刺激HaCaT细胞分泌IL-6和IL-23的影响

Clinical significance of IL-17 expression in serum of patients with psoriasis vulgaris and effects of shikonin on IL-6 and IL-23 production in IL-17-stimulated HaCaT cells

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