circ_0086376靶向miR-5195-3p对痤疮丙酸杆菌介导的HaCaT细胞炎症因子表达的调控作用研究  

Circ_0086376 targeted miR-5195-3p to regulate the expression of inflammatory factors mediated by Propionibacterium acnes in HaCaT cells

作　　者：叶瑞贤 薛如君 梁景耀 张锡宝 Ye Ruixian;Xue Rujun;Liang Jingyao;Zhang Xibao(Guangzhou Dermatology Hospital,Institute of Dermatology,Guangzhou Medical University,Guangzhou 510095,China)

机构地区：[1]广州市皮肤病医院、广州医科大学皮肤病研究所,广州510095

出　　处：《中华皮肤科杂志》2025年第4期340-346,共7页Chinese Journal of Dermatology

基　　金：广东省医学科学技术研究基金(A2022052)、广州市科技计划(2023A03J0944)。

摘　　要：目的探讨环状RNA circ_0086376对微小RNA miR-5195-3p的调控作用及对痤疮相关炎症因子表达的影响。方法构建过表达和干扰circ_0086376的HaCaT稳定细胞株,与痤疮丙酸杆菌(P.acne)共培养,采用实时荧光定量(qRT)PCR检测共培养对circ_0086376表达的影响,酶联免疫吸附试验(ELISA)检测细胞培养上清液中炎症因子的表达。将过表达或干扰circ_0086376质粒、模拟或抑制miR-5195-3p片段的慢病毒转染HaCaT细胞,采用ELISA检测过表达或干扰circ_0086376及与P.acne共培养对HaCaT细胞培养上清液中炎症因子的表达。采用荧光素酶活性实验验证circ_0086376与miR-5195-3p的靶向性。采用ELISA检测过表达/干扰circ_0086376与模拟/抑制miR-5195-3p对与P.acne共培养的HaCaT细胞培养上清液中炎症因子表达的影响。结果P.acne与HaCaT细胞共培养后,培养上清液中下述炎症因子水平均高于单纯HaCaT细胞组,白细胞介素(IL)1β[(355.80±23.20)比(260.50±16.58)pg/ml,t=5.79,P<0.01]、IL-6[(38.04±2.69)比(14.33±0.75)pg/ml,t=14.65,P<0.01]、IL-12[(10.87±0.78)比(6.52±0.77)pg/ml,t=6.89,P<0.01]、IL-18[(222.60±21.07)比(146.10±9.14)pg/ml,t=5.77,P<0.01]和肿瘤坏死因子α(TNF-α)[(50.39±1.29)比(20.46±0.83)pg/ml,t=33.83,P<0.01]。circ空载体+P.acne组HaCaT细胞培养上清液中IL-1β、IL-6、IL-12、IL-18和TNF-α表达水平高于过表达circ空载体组,circ过表达+P.acne组上述炎症因子水平低于过表达circ空载体+P.acne组(均P<0.01);干扰circ空载体+P.acne组HaCaT细胞培养上清液中IL-1β、IL-6、IL-12、IL-18和TNF-α表达水平高于干扰circ空载体组,干扰circ+P.acne组高于干扰circ空载体+P.acne组(均P<0.01)。荧光素酶活性实验证实circ_0086376可与miR-5195-3p结合。circ过表达组细胞IL-1β、IL-6、IL-12、IL-18和TNF-α表达水平低于空载体组(均P<0.05),circ过表达+miR模拟组上述炎症因子水平高于circ过表达组(均P<0.05);干扰circ组细胞各炎症因子表达水平高于空载�Objective To investigate the effects of circular RNA circ_0086376 on the expression of acne-related inflammatory factors by targeting microRNA(miR)-5195-3p.Methods Circ_0086376-overexpressing or-interferred stable HaCaT cell lines were constructed and co-cultured with Propionibacterium acnes(P.acne).Quantitative real-time PCR was used to detect the effect of co-culture on circ_0086376 expression,while enzyme-linked immunosorbent assay(ELISA)was performed to measure the expression of inflammatory factors in the cell culture supernatant.The overexpression or interference of circ_0086376 plasmid and lentivirus mimicking or inhibiting miR-5195-3p fragment were transfected into HaCaT cells,and ELISA was used to detect the expression of inflammatory factors in the culture supernatant of HaCaT cells after overexpression or interference of circ_0086376 cultured alone or with P.acne.The luciferase reporter assay was conducted to verify the targeting relationship between circ_0086376 and miR-5195-3p.Additionally,ELISA was used to detect the effects of overexpression/interference of circ_0086376 and mimic/inhibition of miR-5195-3p on the expression of inflammatory factors in the culture supernatant of HaCaT cells co-cultured with P.acne.Results After co-culture with P.acne,the levels of inflammatory factors in the culture supernatant were significantly higher than those in the HaCaT cells cultured alone,including Interleukins(IL)-1β([355.80±23.20]vs.[260.50±16.58]pg/ml,t=5.79,P<0.01),IL-6([38.04±2.69]vs.[14.33±0.75]pg/ml,t=14.65,P<0.01),IL-12([10.87±0.78]vs.[6.52±0.77]pg/ml,t=6.89,P<0.01),IL-18([222.60±21.07]vs.[146.10±9.14]pg/ml,t=5.77,P<0.01),and Tumor necrosis factor(TNF)-α([50.39±1.29]vs.[20.46±0.83]pg/ml,t=33.83,P<0.01).The expression levels of IL-1β,IL-6,IL-12,IL-18 and TNF-αin the culture supernatant of HaCaT cells in the circ-empty vector+P.acne group were higher than those in the circ-empty vector overexpression group.The levels of inflammatory factors mentioned above in circ-overexpression+P.acne gro

关 键 词：RNA 环状 微RNAS 竞争性内源性RNA 痤疮丙酸杆菌 HACAT细胞 白细胞介素类 肿瘤坏死因子α 

分 类 号：R758.72[医药卫生—皮肤病学与性病学]