清肝凉血解毒汤对咪喹莫特诱导小鼠银屑病样模型皮损的干预作用

Eeffectsof Qinggan Liangxue Jiedu Decoction on Mice Psoriasis-Like Lesions Induced by Imiquimod

目的:观察清肝凉血解毒汤对咪喹莫特诱导的银屑病样小鼠模型皮损的影响。方法:72只BALB/c雄性小鼠,背部刮毛,随机分为空白对照组(Control,C)、模型组(Model,M)、复方高、中、低剂量组(Q-H、Q-M、Q-L)和甲氨蝶呤组(MTX),5%咪喹莫特乳膏(IMQ)背部涂抹诱导皮肤银屑病样模型。采用银屑病皮损面积和疾病严重程度(Psoriasis area and severity index,PASI)每日进行评分,光镜下观察皮损组织形态学变化及表皮厚度;免疫组化法检测皮损中增殖细胞核抗原(PCNA)和T淋巴细胞表面标志CD3表达情况;免疫荧光法检测皮损中CD11c+树突状细胞、CGRP、NK1R表达情况;采用实时PCR技术检测皮损中IL-1β、IL-12、IL-23mRNA表达水平;Western Blot法检测皮损中NPY、SP蛋白表达情况。结果:M组小鼠皮损上鳞屑厚,浸润明显,红斑重;其余各组皮损均较M组轻。与C组相比,M组表皮厚度、PCNA、CD^+_3 T细胞及CD11c+细胞均明显增多(P<0.01);复方各剂量组与M组相比,表皮薄,PCNA、CD^+_3及CD11c+细胞表达个数均明显减少。M组与C组相比,CGRP、NK-1R、IL-1β、IL-12、IL-23mRNA表达均上调(P<0.05),而余组与M组相比,上述指标的表达均明显下调(P<0.05)。与C组相比,M组小鼠皮损中NPY的表达增高(P<0.05),MTX、Q-M、Q-L组表达量较M组低(P<0.05);M、MTX、Q-M、Q-L组皮损中SP的表达均呈增高趋势,但组间相比无统计学差异。结论:清肝凉血解毒汤可通过下调NK-1R、NPY、CGRP的表达水平改善小鼠银屑病样皮损,降低表皮细胞的异常增殖、减轻炎症细胞的浸润,减少皮肤中CD11c+树突状细胞数量,下调IL-1β、IL-12、IL-23细胞因子的表达水平。

Objective:To observe the effects ofQinggan Liangxue Jiedu Decoction(QGLXJD)on mice psoriasis-like lesions induced by imiquimod.Methods:Seventy-two BALB/c mice were randomly divided into 6 groups:control group,model group,QGLXJD groups and MTX group.IMQ was daily used to induce the psoriasis-like lesion model.The model group was given purified water.QGLXJD groups were given QGLXJD.MTX group was givenMethotrexate.The lesions are evaluated according to the psoriasis area and severity index(PASI)daily.On the 7 th day,the histology and epidermal thicknesses were observed under high microscope.Meanwhile,the expressions of proliferating cell nuclear antigen(PCNA),CD3^+T cells,CD11 c+dendritic cells,NK-1 R,CGRP were counted.The expressions of NPY and SP protein were detected by Western blot and IL-1β,IL-12,IL-23 detected by PCR.Results:The skin lesions of the model group were thick,infiltrateand erythema.Compared with the model group,the skin lesions of the other groups were relatively light.Compared with the control group,the epidermal value of the model group was significantly increased,and the PCNA,CD3 and CD11 c positive cells were significantly increased(P<0.01).Compared with the model group,the expressions of PCNA,CD3 and CD11 c positive cells decreased significantly in other groups.Compared with the control group,CGRP and NK-1 R expressions in model group were raised(P<0.01),while the treatment group’s were significantly lowered(P<0.05)when compared with those of the model group.Compared with the model group,the relative expression levels of IL-1,IL-12 and IL-23 mRNA were significantly decreased in the other groups.The expression of NPY in the model group was significantly higher than that in the other groups(P<0.05).Compared with the control group,the expression of SP in the skin lesions of psoriatic groups showed an increasing trend.Conclusion:QGLXJD can improve the mice psoriasis-like skin through down-regulating the expressions of NK-1 R,NPY and CGRP,reducing the abnormal epidermal cell proliferation,inflammation cells infiltration,CD11 c+dendritic cellsand the expressions of IL-12,IL-23 and IL-1βcytokine.