黄芩素通过抑制NF-κB通路对小鼠烧伤愈合的促进作用

Effects of baicalein on promoting burn healing in mice via inhibiting NF-κB pathway

目的 探究黄芩素对小鼠烧伤愈合的作用,并阐明其机制。方法 分离成年小鼠原代皮肤成纤维细胞经热刺激30 s后,分别以20、50μmol/L黄芩素处理细胞;经热刺激后的细胞或以50μmol/L黄芩素单独处理后与100 ng/mL TNF-α重组蛋白共处理细胞。采用流式细胞术、CCK-8、Western blot法分别检测细胞凋亡、增殖、蛋白表达。将小鼠随机分为对照组,烧伤组,20、50 mg/kg黄芩素组,黄芩素(50 mg/kg)+TNF-α重组蛋白(5μg/kg)组,每隔2 d测量伤口面积,在第8天进行组织学检查和烧伤组织中凋亡、炎症相关蛋白表达检测。结果 黄芩素处理可抑制热刺激诱导的细胞凋亡和炎症,促进细胞增殖(P<0.01),而TNF-α处理阻碍黄芩素对热刺激诱导的细胞损伤的抑制作用(P<0.01)。黄芩素腹腔注射可促进烧伤愈合,降低烧伤组织中的炎症和细胞凋亡(P<0.01);TNF-α处理可阻碍黄芩素对烧伤愈合的促进作用(P<0.01)。结论 黄芩素通过抑制NF-κB通路缓解热刺激对原代小鼠皮肤成纤维细胞的损伤,促进小鼠烧伤愈合。

AIM To investigate the effects of baicalein on mouse burn healing and elucidate its mechanism.METHODS The isolated primary skin fibroblasts of adult mice were treated with 20, 50 μmol/L baicalein respectively following 30 s thermal stimulation. The other cells exposed to the heat were co-treated with 50 μmol/L baicalein and 100 ng/mL TNF-α recombinant protein. These cells had their cell apoptosis, proliferation and protein expressions detected by flow cytometry, CCK-8 and Western blot. The mice were randomly divided into the control group, burn group, 20, 50 mg/kg baicalein group, baicalein(50 mg/kg) + TNF-α recombinant protein(5 μg/kg) group. The mice had their wound size measured every two days, and histological indices levels and expressions of apoptosis and inflammation-related proteins in the burn tissues determined on the eighth day.RESULTS Baicalein treatment inhibited thermal stimulation-induced apoptosis and inflammation, and promoted cell proliferation(P<0.01) which was suppressed by TNF-α treatment(P<0.01). Intraperitoneal injection of baicalein promoted burn healing and reduced inflammation and apoptosis in the burn tissue(P<0.01), which was offset by the injection of TNF-α(P<0.01).CONCLUSION Baicalin alleviates the damage of primary mice skin fibroblasts induced by thermal stimulation, and promotes burn healing in mice via inhibiting NF-κB signaling pathway.