Department of Dermatology, Qilu Hospital, Shangdong University, 107 West Wenhua Road, Jinan, Shandong, 250012, China.
Inflammation. 2018 Mar;41(2):496-504. doi: 10.1007/s10753-017-0705-3.
Psoriasis is an immune-mediated chronic skin disorder where T cells play a main role, and numerous inflammatory cytokines are implicated in its pathogenesis by initiating keratinocyte proliferation. Interleukin-22 (IL-22), an IL-10 family cytokines, is critical in the pathogenesis and development of psoriasis. To determine the target of microRNA (miR) -548a-3p and investigate its role in keratinocyte proliferation after treating human keratinocytes (HaCaT) with IL-22, we used quantitative reverse transcriptase PCR to measure the expression of miR-548a-3p in both HaCaT cells stimulated with IL-22 and psoriatic lesions, and then detected the biological function of miR-548a-3p in HaCaT cells by performing Counting Kit-8 (CCK-8) assays. Luciferase reporter assay was conducted to determine the target gene of miR-548a-3p. Immunohistochemistry and Western blot were performed to verify the target gene. Results showed that miR-548a-3p was significantly upregulated both in HaCaT cells treated with IL-22 and psoriatic lesions. The over expression of miR-548a-3p could promote the proliferation of HaCaT cells. Luciferase was mutated in the 3'UTR of PPP3R1, a gene coding Calcineurin. Immunohistochemistry and Western blot demonstrated that the expression of PPP3R1 decreased respectively in psoriatic lesions and HaCaT cells. In conclusion, the expression of miR-548a-3p is upregulated in IL-22 mediated keratinocyte proliferative disorder like psoriasis. The impact of miR-548a-3p on keratinocyte proliferation may be implemented by targeting PPP3R1 and T regulatory cells may be involved in the pathogenesis of psoriasis.
银屑病是一种免疫介导的慢性皮肤疾病，其中 T 细胞发挥主要作用，许多炎症细胞因子通过启动角质形成细胞增殖而参与其发病机制。白细胞介素-22（IL-22）是一种白细胞介素-10 家族细胞因子，在银屑病的发病机制和发展中起关键作用。为了确定 microRNA（miR）-548a-3p 的靶标，并研究其在人角质形成细胞（HaCaT）经 IL-22 处理后增殖的作用，我们使用定量逆转录酶 PCR 测量了在 IL-22 刺激的 HaCaT 细胞和银屑病病变中 miR-548a-3p 的表达，然后通过细胞计数试剂盒（CCK-8）测定法检测 miR-548a-3p 在 HaCaT 细胞中的生物学功能。进行荧光素酶报告基因测定以确定 miR-548a-3p 的靶基因。进行免疫组织化学和 Western blot 以验证靶基因。结果表明，miR-548a-3p 在经 IL-22 处理的 HaCaT 细胞和银屑病病变中均显著上调。miR-548a-3p 的过表达可促进 HaCaT 细胞的增殖。PPP3R1 的 3'UTR 中的荧光素酶发生突变，该基因编码钙调磷酸酶。免疫组织化学和 Western blot 表明 PPP3R1 的表达分别在银屑病病变和 HaCaT 细胞中降低。总之，miR-548a-3p 的表达在像银屑病这样的 IL-22 介导的角质形成细胞增殖障碍中上调。miR-548a-3p 对角质形成细胞增殖的影响可能通过靶向 PPP3R1 来实现，调节性 T 细胞可能参与银屑病的发病机制。
