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

doi:10.16352/j.issn.1001-6325.2023.01.0123

摘要/Abstract

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、150 μg/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。

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

Abstract: 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, 150 μg/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-α and 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.

Key words: psoriasis, Scutellaria extract, miR-369-3p, IL-22, human keratinocytes, inflammatory reaction

中图分类号:

R310.17

曾颖, 付桂莉. 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[J]. 基础医学与临床, 2023, 43(1): 123-129.

ZENG Ying, FU Guili. Scutellaria Extract inhibits IL-22-induced hyperproliferation of human keratinocyte cell line HaCaT[J]. Basic & Clinical Medicine, 2023, 43(1): 123-129.

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