Clascoterone对SZ95人皮脂腺细胞增殖、脂质合成和炎症因子表达的影响

Effect of clascoterone on the proliferation, lipid synthesis and inflammatory factors expression of human SZ95 sebocytes in vitro

目的:明确clascoterone对雄激素作用下人皮脂腺细胞增殖、脂质合成和炎症因子表达的影响,探索其抗痤疮机制。方法:体外培养SZ95人皮脂腺细胞,分为双氢睾酮(DHT)组、clascoterone组,clascoterone+DHT组,CCK8法检测细胞增殖,尼罗红染色检测细胞内中性脂质,蛋白免疫印迹检测脂质合成相关基因表达。酶联免疫吸附检测细胞培养上清液中炎症因子蛋白,RT-qPCR检测细胞因子转录水平。结果:与DHT组相比,clascoterone和DHT共培养24h后细胞数量出现显著下降(P<0.01);在亚油酸(LA)作用下,clascoterone+DHT组细胞内中性脂质低于DHT组(P<0.0001);此外,clascoterone+DHT组PPARγ、PLIN2、FASN、SCD1等脂质合成相关基因的表达、炎症因子IL-1α、IL-1β、IL-6、IL-8及相应蛋白水平低于DHT组(均P<0.05)。结论:clascoterone体外对人皮脂腺细胞增殖、脂质合成相关基因和炎症因子具有调节作用,可能是其抗痤疮治疗机制之一。

Objective: To determine the effect of clascoterone on androgen-induced human sebocytes proliferation, lipid synthesis and inflammatory factors expression in vitro and explore its anti-acne mechanism. Methods: The immortalized human sebocytes SZ95 were cultrured in vitro and then divided into Dihydrotestosterone(DHT) group, clascoterone group and clascoterone+DHT group. The cell proliferation, intracellular neutral lipids, lipid synthesis-related proteins expression, inflammatory factors and transcription level of cytokines was detected by CCK8, nile red staining and western blot, enzyme-linked immunosorbent assay and RT-qPCR respectively. Results: Compared with the DHT group, the number of cells in the clascoterone+DHT group significantly decreased(P<0.01) after 24-hour treatment. After treated with linoleic acid(LA), the level of neutral lipids in the clascoterone+DHT group was significantly lower than that in the DHT group(P<0.0001). The lipid synthesis-related proteins such as PPARγ, PLIN2, FASN and SCD1, the transcription levels of IL-1α, IL-1β, IL-6, IL-8 and the corresponding protein levels in the clascoterone+DHT group was significantly lower than those in the DHT group(P;<0.05). Conclusion: Clascoterone has a partly androgen-independent regulatory effect on human sebocytes proliferation, lipid synthesis-related genes and inflammatory factors in vitro, which might be one of anti-acne mechanisms.