黄芩提取物抑制IL⁃22诱导的人角质形成细胞系HaCaT过度增殖  

Scutellaria Extract inhibits IL-22-induced hyperproliferation of human keratinocyte cell line HaCaT

作　　者：曾颖 付桂莉[1] ZENG Ying;FU Guili(Department of Dermatology,Wuhan Children's Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430019,China)

机构地区：[1]华中科技大学同济医学院附属武汉儿童医院皮肤科,湖北武汉430019

出　　处：《基础医学与临床》2023年第1期123-129,共7页Basic and Clinical Medicine

基　　金：武汉市医学科研项目(WX19Y01)。

摘　　要：目的探究黄芩提取物通过miR-369-3p对白介素-22(IL-22)诱导的人角质形成细胞系HaCaT增殖、凋亡和炎性反应的影响。方法将HaCaT细胞分为对照组、IL-22组(细胞模型,100 ng/mL的IL-22培养细胞)、低、中和高剂量组(黄芩提取物50、100、150μg/mL)、anti-miR-NC干预组(转染anti-miR-NC)、anti-miR-369-3p干预组(转染anti-miR-369-3p)、miR-NC+高剂量组(转染miR-NC,高剂量黄芩提取物培养细胞)、miR-369-3p+高剂量组(转染miR-369-3p,高剂量黄芩提取物培养细胞)。MTT法检测细胞增殖;流式细胞测量术检测细胞凋亡;Western blot检测cleaved caspase-3蛋白表达;ELISA检测TNF-α、IL-6含量;RT-qPCR检测miR-369-3p表达。结果与对照组相比,IL-22组细胞存活率、miR-369-3p表达、TNF-α、IL-6表达升高(P<0.05),细胞凋亡率、cleaved caspase-3蛋白表达的差异无统计学意义;与IL-22组相比,低、中和高剂量组细胞存活率、miR-369-3p表达、TNF-α、IL-6表达降低,细胞凋亡率、cleaved caspase-3蛋白表达升高(P<0.05)。与anti-miR-NC干预组相比,anti-miR-369-3p干预组细胞存活率、TNF-α、IL-6表达降低,细胞凋亡率、cleaved caspase-3蛋白表达升高(P<0.05)。过表达miR-369-3p可以部分逆转黄芩提取物对银屑病细胞模型的细胞活性,凋亡以及炎性水平的作用(P<0.05)。结论黄芩提取物可能通过抑制miR-369-3p表达,抑制IL-22诱导的人角质形成细胞过度增殖和炎性反应,并促进其凋亡。Objective To explore the experimental study of Scutellaria extract on IL-22-induced proliferation,apoptosis and inflammatory response of human keratinocyte cells line(HaCaT)via miR-369-3p.Methods HaCaT cells were divided into control group,IL-22 group(cell models,100 ng/mL of IL-22 cultured cells),low,medium and high dose group(Scutellaria extract 50,100,150μg/mL),anti-miR-NC intervention group(transfected anti-miR-NC),anti-miR-369-3p intervention group(transfected with anti-miR-369-3p),miR-NC+high-dose group(transfected with miR-NC,Scutellaria extract cultured cells),miR-369-3p+high-dose group(transfected with miR-369-3p,Scutellaria extract cultured cells).MTT experiment detected cell proliferation;Flow cytometry detected cell apoptosis;Western blot detected cleaved caspase-3 protein expression;ELISA detected TNF-αand IL-6 content;RT-qPCR detected miR-369-3p expression.Results Compared with the control group,the IL-22 group cell survival rate,miR-369-3p expression,TNF-α,IL-6 expression increased(P<0.05),and the difference in the cell apoptosis rate,cleaved caspase-3 protein expression were not signficantly(P>0.05);Compared with the IL-22 group,the cell survival rate,miR-369-3p expression,TNF-α,IL-6 expression decreased in the low,middle and high dose groups,while the apoptosis rate,cleaved caspase-3 protein expression increased(P<0.05).Compared with the anti-miR-NC intervention group,the anti-miR-369-3p intervention group had lower cell viability,TNF-α,IL-6 expression,while apoptosis rate,cleaved caspase-3 protein expression increased(P<0.05).Over-expression of miR-369-3p could partially reverse the effects of Scutellaria extract on cell viability,apoptosis and inflammatory response of psoriasis cell models.Conclusions Scutellaria extract inhibits IL-22-induced hyperproliferation and inflammatory response of human keratinocytes,and promotes apoptosis with potential mechanisms by inhibiting the expression of miR-369-3p.

关 键 词：银屑病 黄芩提取物 miR-369-3p IL-22 人角质形成细胞 炎性反应 

分 类 号：R310.17[医药卫生—基础医学]