Circ_0086376 targeted miR-5195-3p to regulate the expression of inflammatory factors mediated by Propionibacterium acnes in HaCaT cells
- VernacularTitle: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
- Author:
Ruixian YE
1
;
Rujun XUE
1
;
Jingyao LIANG
1
;
Xibao ZHANG
1
Author Information
- Publication Type:Journal Article
- Keywords: RNA, circular; MicroRNAs; Competitive endogenous RNA; Propionibacterium acnes; HaCaT cells; Interleukins; Tumor necrosis factor-alpha
- From: Chinese Journal of Dermatology 2025;58(4):340-346
- CountryChina
- Language:Chinese
- Abstract: 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-α in 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 group were lower than those in circ-overexpression empty vector + P.acne group ( P < 0.01). The expression levels of inflammatory factors mentioned above in the culture supernatant of HaCaT cells in the interference circ-empty vector + P.acne group were higher than those in the interference circ-empty vector group. Compared with the interference circ-empty vector + P.acne group, the expression of circ in the interference circ + P.acne group was higher ( P < 0.01). Luciferase reporter assay confirmed that circ_0086376 could bind to miR-5195-3p. The expression levels of IL-1β, IL-6, IL-12, IL-18 and TNF-α in the circ-overexpression group were lower than those in the empty vector group ( P < 0.05), and the levels of these inflammatory factors in the circ-overexpression + miR mimic group were higher than those in the circ overexpression group ( P < 0.05). The expression levels of inflammatory factors in the interference circ group were higher than those in the empty vector group, levels of inflammatory factors in the circ + miR-inhibiting group were lower than those in the circ interference group ( P < 0.05) . Conclusion:Circ_0086376 can inhibit the expression of acne-related inflammatory factors by targeting and downregulating miR-5195-3p.
