紫草素联合当归饮子方调控IL-23/IL-17轴对咪喹莫特诱导银屑病样小鼠模型的保护作用

Protective Effect of Shikonin Combined with Dangguiyinzi Decoction on Imiquimote-induced Psoriasis in Mice by Regulating IL-23/IL-17 Axis

目的探讨紫草素联合当归饮子方通过调控IL-23/IL-17轴对咪喹莫特诱导的银屑病样小鼠模型的保护作用及机制。方法将30只KM雄性小鼠随机分为空白对照组、模型组、紫草素治疗组、当归饮子治疗组、紫草素+当归饮子治疗组,每组6只。在等面积(2 cm×3 cm)小鼠背部皮肤上除去被毛,除空白对照组外其余各组小鼠涂抹咪喹莫特乳膏(62.5 mg/只),紫草素治疗的小鼠涂抹紫草素0.5 mL;当归饮子治疗的当归饮子灌胃4 mL/(kg·d),1次/d,连续7 d,期间观察并记录各组小鼠背部PASI评分。在第8天麻醉后处死小鼠并收集血清及小鼠皮损组织,HE染色观察表皮厚度,ELISA法检测各组小鼠皮损组织及血清中IL-17A、IL-6、IL-23A和IL-1β水平,qRT-PCR方法检测各组皮损中IL-17A、IL-6、IL-23A和IL-1β的基因表达情况。结果PASI评分及HE染色结果显示,在紫草素联合当归饮子给药第7天可显著改善小鼠背部皮肤发红、增厚、皮屑增多等银屑病特征性临床表现,小鼠背部皮肤结痂及表皮增厚现象明显减轻;在皮损组织及血清样本中,模型组IL-17A、IL-23A、IL-6和IL-1β水平较对照组明显上升(P<0.0001),各治疗组IL-17A、IL-23A、IL-6和IL-1β水平较模型组均明显降低,紫草素+当归饮子治疗组IL-23A、IL-6水平显著低于当归饮子治疗组,IL-17A水平显著低于当归饮子治疗组及紫草素治疗组;模型组小鼠皮损组织中IL-17A、IL-23A、IL-6和IL-1βmRNA表达水平显著高于对照组(P<0.0001),各治疗组IL-17A、IL-23A、IL-6和IL-1βmRNA表达水平较模型组均明显降低,紫草素+当归饮子治疗组IL-23A、IL-17A mRNA表达水平显著低于当归饮子治疗组。结论紫草素与当归饮子方联合应用可调节IL-23/IL-17轴,从而降低银屑病小鼠血清及皮损组织IL-6、IL-23A、IL-1β及IL-17A的表达。

Objective To explore the protective effect of the combination of shikonin and Dangguiyinzi decoction in treating imiquimod-induced psoriasis in mice through the regulation of the IL-23/IL-17 axis.Methods Thirty male KM mice were randomly assigned to 5 groups(n=6),namely the control group(Control),the model group(Model),the shikonin group(Shikonin),the Dangguiyinzi group(DGYZ),and the combination group(Shikonin+DGYZ).Fur was removed from the dorsal area(2 cm×3 cm)of all mice,imiquimod(62.5 mg each)was applied in all groups except for the control group.The shikonin-treated mice were treated with 0.5 mL of shikonin.The DGYZ-treated mice were given Dangguiyinzi decoction at a dose of 4 mL/(kg·d)via gastric lavage once daily for seven consecutive days,and the Psoriasis Area and Severity Index(PASI)score of the dorsal area of each group was observed and recorded.On day 8,the mice were killed after anesthesia to collect serum and skin samples.HE staining was conducted to observe the thickness of the epidermis,ELISA was performed to determine IL-17 A,IL-6,IL-23 A,and IL-1βlevels in skin tissue and serum,and quantitative real-time PCR(qRT-PCR)was implemented to analyze IL-17 A,IL-6,IL-23 A,and IL-1βgene expression levels in the skin.Results The PASI scores and HE staining showed that in the combination group,distinct clinical features of psoriasis such as redness,thickened skin,and increased silvery scales were ameliorated at Day 7.Scrabs and epidermal thickening on the dorsal area were reduced significantly.IL-17 A,IL-23 A,IL-6,and IL-1βgene and serum levels were significantly increased in the model group(P<0.0001)while,in sharp contrast,they were significantly reduced in the treatment group.In the Shikonin+DGYZ group,the IL-23 A and IL-6 levels were significantly lower than in the DGYZ group,and the IL-17 A level was markedly lower than in the DGYZ group and the Shikonin group.The IL-17 A,IL-23 A,IL-6,and IL-1βmRNA expression levels in skin tissue were substantially higher in the model group compared with the control group(P<0.0001).Compared with the model group,the treatment group had significantly reduced IL-17 A,IL-23 A,IL-6,and IL-1βmRNA expression levels.The IL-23 A and IL-17 A mRNA expression levels in the Shikonin+DGYZ group were remarkably lower than in the DGYZ group.Conclusion The combination of shikonin and Dangguiyinzi decoction can regulate the IL-23/IL-17 axis,thereby reducing the expression of IL-6,IL-23 A,IL-1β,and IL-17 A in serum and skin tissue of psoriatic mice.