基于TWEAK/Fn-14信号通路探讨半枝莲方治疗银屑病的机制研究

Study on the mechanism of Banzhilian formula in the treatment ofpsoriasis based on TWEAK/Fn-14 signaling pathway

目的 探讨半枝莲方对咪喹莫特诱导的银屑病样小鼠模型皮损组织中肿瘤坏死因子样凋亡诱导物(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)、成纤维细胞生长诱导因子(fibroblast growth factor-inducible,Fn)-14以及脂质运载蛋白(lipocalin,Lcn)-2表达水平影响,并探讨其作用机制。方法 将21只BALB/c雄性小鼠随机分为对照组、模型组以及半枝莲方干预组,每组7只。对照组外涂凡士林及生理盐水灌胃,模型组外涂咪喹莫特及生理盐水灌胃,干预组外涂咪喹莫特及半枝莲方灌胃,连续7 d。比较各组小鼠银屑病皮损面积和严重程度指数(psoriasis area and severity index,PASI)评分;苏木精-伊红染色观察皮损组织病理改变;免疫组织化学法检测皮损组织中Ki-67、Fn-14以及Lcn-2表达;采用蛋白质印迹法检测皮损组织Fn-14、Lcn-2水平;酶联免疫吸附试验测定皮损中TWEAK含量;定量聚合酶链反应检测皮损组织TWEAK、Fn-14以及Lcn-2 mRNA表达。结果 与模型组相比,半枝莲方干预组小鼠皮肤红斑、鳞屑严重程度减轻,PASI评分下降,Ki-67阳性细胞数减少,同时显著降低皮损组织中TWEAK、Fn-14以及Lcn-2 mRNA和蛋白表达(P<0.05)。结论 半枝莲方能够改善咪喹莫特诱导的小鼠银屑病样模型皮肤炎症反应,可能是通过抑制TWEAK/Fn-14信号通路发挥作用。

Objective To explore the effects of Banzhilian formula on the expression levels of tumor necrosis factorlike weak inducer of apoptosis(TWEAK),fibroblast growth factor-inducible(Fn)-14,and lipocalin(Lcn)-2 in the psoriatic mouse skin tissue induced by imiquimod,and to investigate the mechanism of action.Method A total of 21 BALB/c male mice were randomly divided into control group,model group and intervention group,with 7 mice in each group.The control group was given Vaseline and 0.9%sodium chloride injection intragastric administration,the model group was given imiquimod and 0.9%sodium chloride injection intragastric administration,and the intervention group was given imiquimod and Banzhilian formula intragastric administration for 7 consecutive days.The psoriasis area and severity index(PASI)scores in each group were compared.Hematoxylin eosin staining was used to observe the pathological changes of skin tissue. The expressions of Ki-67, Fn-14 and Lcn-2 in skin lesions were detected by immunohistochemistry. The levelsof Fn-14 and Lcn-2 in skin lesions were detected by western blot. TWEAK content in skin lesions was determined byenzyme linked immunosorbent assay. The mRNA expression of TWEAK, Fn-14 and Lcn-2 were detected by quantitativepolymerase chain reaction. Results Compared with the model group, the severity of skin erythema and scales in theintervention group were reduced, PASI score and the number of Ki-67 positive cells were decreased, the mRNA andprotein expression of TWEAK, Fn-14, and Lcn-2 in the lesion tissues were significant decreased (P<0.05). ConclusionBanzhilian formula can improve imiquimod-induced skin inflammatory response in mouse psoriasis model, possibly byinhibiting TWEAK/Fn-14 signaling pathway.