miR-128-3p inhibits the proliferation of keratinocytes in psoriasis via repressing leptin
10.3969/j.issn.1674-8115.2024.10.005
- VernacularTitle:miR-128-3p靶向沉默瘦素抑制银屑病角质形成细胞增殖及炎症反应
- Author:
Jing PENG
1
;
Jing YIN
;
Ping XIA
;
Liuqing CHEN
Author Information
1. 湖北省武汉市第一医院皮肤科,皮肤感染和免疫湖北省重点实验室,武汉 430022
- Keywords:
psoriasis;
leptin;
miR-128-3p;
keratinocyte
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2024;44(10):1241-1248
- CountryChina
- Language:Chinese
-
Abstract:
Objective·To explore the role of miR-128-3p/leptin(LEP)axis in the proliferation and inflammation of keratinocytes in psoriasis.Methods·BALB/c mice were randomly divided into a control group(n=10)and a model group(n=10).Mice in the model group were given imiquimod on the back.miR-128-3p overexpression and interference plasmids,as well as LEP interference plasmids,were constructed and transfected into HaCaT cells,respectively.miR-128-3p and LEP mRNA were quantified by real-time quantitative polymerase chain reaction,and LEP protein levels were detected by using Western blotting.Enzyme-linked immunosorbent assay was used to measure the content of tumor necrosis factor α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in the culture medium.MTT assay was used to evaluate cell activity and EdU assay was to used to test cell proliferation.The binding site between miR-128-3p and LEP was determined by using a dual luciferase reporter gene assay.Results·Compared with mice in the control group,mice in the model group showed downregulated expression of miR-128-3p and upregulated expression of LEP at both RNA and protein levels(all P＜0.05).The dual luciferase reporter gene assay confirmed that LEP was a downstream target of miR-128-3p.Compared with the negative control mimic(NC mimic)group,expression of miR-128-3p was up-regulated in the miR-128-3p mimic group,and expression of LEP was reduced.The levles of TNF-α,IL-6,and IL-1β were significantly lower in the miR-128-3p mimic group than in the NC mimic group.The relative cell viability and EdU-positive cell rate were also reduced after miR-128-3p up-regulation(all P＜0.05).Compared with the negative control inhibitor(NC inhibitor)group,expression of miR-128-3p was down-regulated in the miR-128-3p inhibitor group,and expression of LEP was increased.The levles of TNF-α,IL-1β and IL-6 were increased after miR-128-3p downregulation.miR-128-3p down-regulation led to an increase in relative cell viability and EdU-positive cell rate(all P＜0.05).Further experimental results showed that LEP expression was up-regulated in the miR-128-3p inhibitor+LEP inhibitor group compared with that in the LEP inhibitor group,whereas the levels of TNF-α,IL-6,and IL-1β were elevated,and the relative viability of the cells and the rate of EdU-positive cells were increased(all P＜0.05).Conclusion·miR-128-3p downregulates LEP to inhibit the proliferation and inflammatory response of keratinocytes,thereby inhibiting the occurrence and development of psoriasis.
