清血毒胶囊对银屑病模型小鼠的治疗效果及皮损部HDAC1、Gli1蛋白表达的影响

Effect of Qingxuedu Capsule(清血毒胶囊)on the Therapeutic Effect of Psoriasis Model Mice and the Expression of HDAC1 and Gli1 Proteins in Skin Lesions

目的:观察清血毒胶囊对银屑病模型小鼠治疗效果及组蛋白脱乙酰基酶1(HDAC1)、胶质瘤相关癌基因蛋白-1(Gli1)表达的影响。方法:60只BALB/c小鼠随机分为空白组、模型组、甲氨蝶呤组、清血毒胶囊低剂量组、清血毒胶囊中剂量组和清血毒胶囊高剂量组,每组10只。采用5%咪喹莫特软膏制备寻常型银屑病小鼠模型,同时予治疗药物干预,各组均连续干预7 d。观察记录各组小鼠(第1天、第3天、第7天)皮损颜色、鳞屑程度、皮肤厚度并予银屑病皮损严重程度指数评分(Psoriasis Area and Severity Index,PASI);干预结束后,取皮损组织通过苏木精-伊红(HE)染色检测皮肤组织病理学变化并予Baker法评分;蛋白免疫印迹法检测皮损部HDAC1和Gli1的蛋白表达。结果:不同时间(第1天、第2天、第3天)之间小鼠PASI评分表达比较,差异有统计学意义(P<0.05),即存在时间效应。除空白组外,其余各组小鼠的PASI评分均逐渐上升。6组小鼠PASI评分总体比较,差异有统计学意义(P<0.05),即存在分组效应。时间因素和组别因素对PASI评分的影响存在交互效应(P<0.05)。干预7 d后,与空白组比较,模型组小鼠PASI评分、Baker法评分及HDAC1和Gli1的蛋白相对表达量均明显升高(P<0.05);与模型组比较,甲氨蝶呤组和清血毒胶囊中、高剂量组小鼠PASI评分、Baker法评分及HDAC1蛋白相对表达量均明显降低(P<0.05),清血毒胶囊低、中、高剂量组Gli1蛋白表达相对表达量均明显降低(P<0.05),且与药物浓度成负相关。结论:清血毒胶囊干预银屑病小鼠疗效肯定,其可能通过抑制HDAC1的表达,调控hedgehog(Hh)信号通路转录因子Gli1表达,抑制银屑病角质形成细胞异常增殖,发挥干预银屑病的作用。

Objective:To observe the therapeutic effect of Qingxuedu Capsule on psoriasis model mice and the expression of histone deacetylase 1(HDAC1)and glioma associated oncogene protein-1(Gli1).Methods:Sixty BALB/c mice were randomly divided into blank group,model group,Methotrexate group and Qingxuedu Capsule low,medium and high dose groups,with 10 mice in each group.The mouse model of psoriasis vulgaris was prepared with 5%imiquimod ointment and treated with therapeutic drugs for 7 days.Observe and record the skin lesion color,scale degree and skin thickness of mice in each group(day 1,day 3 and day 7),and score the psoriatic area and severity index(PASI).After the intervention,the pathological changes of skin tissue were detected by hematoxylin eosin(HE)staining and scored by Baker method.The protein expression of HDAC1 and Gli1 was detected by Western blot.Results:The expression of PASI score in mice at different times(day 1,day 2 and day 3)was statistically significant(P<0.05),which means there was a time effect.Except the blank group,the PASI scores of mice in other groups increased gradually.There was significant difference in PASI score between the 6 groups(P<0.05),which means there was a grouping main effect.There was an interaction effect between time factor and group factor on PASI score(P<0.05).After 7 days of intervention,compared with the blank group,the PASI score,Baker method score and the relative expression of HDAC1 and Gli1 protein in the model group were significantly increased(P<0.05).Compared with the model group,the PASI score,Baker method score and the relative expression of HDAC1 protein in the methotrexate group and the middle and Qingxuedu Capsule medium and high dose groups decreased significantly(P<0.05),and the relative expression of Gli1 protein in the Qingxuedu Capsule low,middle and high dose groups decreased significantly(P<0.05),which was negatively correlated with the drug concentration.Conclusion:Qingxuedu Capsule has a positive effect on psoriatic mice.It may inhibit the abnormal proliferation of psoriatic keratinocytes by inhibiting the expression of HDAC1 and regulating the expression of hedgehog(Hh)signal pathway transcription factor Gli1,and play a role in the intervention of psoriasis.