健脾解毒汤对脾虚湿盛证银屑病大鼠皮肤组织AQP3蛋白表达的影响

Effect of decoction for fortifying spleen and removing toxin on AQP3 protein expression in skin tissues of psoriasis rat with syndrome of spleen deficiency and dampness exuberance

目的研究健脾解毒汤对脾虚湿盛证银屑病大鼠皮肤组织水通道蛋白3(AQP3)表达的影响,探讨健脾解毒汤修复皮肤屏障功能的作用机制。方法按照随机数字表将72只雄性SD大鼠随机分为空白组、模型组、甲氨蝶呤组、健脾解毒汤低剂量组、健脾解毒汤中剂量组、健脾解毒汤高剂量组,每组12只。除空白组大鼠外,其余组大鼠采用咪喹莫特乳膏涂抹及大黄配方颗粒剂灌胃方法诱导建立脾虚湿盛证银屑病模型。造模结束后,甲氨蝶呤组给予甲氨蝶呤片0.97 mg/kg灌胃,健脾解毒汤低、中、高剂量组给予健脾解毒汤2.32 g/kg、4.64 g/kg、9.26 g/kg灌胃,均1次/d,连续14 d。观察各组大鼠表征特点,HE染色观察皮肤组织病理形态,Western blot法检测皮肤组织中AQP3蛋白表达情况。结果灌胃结束后,模型组大鼠进食量减少,排便次数增加,粪便稀;健脾解毒汤各组大鼠进食量明显改善,排便次数明显减少,粪便质地适中。HE染色显示模型组大鼠表皮增生肥厚,显著角化不全伴角化过度,表皮突延长,部分表皮细胞间水肿,表皮内及角质层内较多中性粒细胞浸润,真皮浅层内血管扩张,慢性炎细胞散在或灶状浸润;健脾解毒汤各组大鼠皮肤组织结构较模型组明显改善。模型组大鼠皮肤组织中AQP3蛋白表达量明显低于空白组(P<0.05),甲氨蝶呤组和健脾解毒汤各组大鼠AQP3表达量均明显高于模型组(P均<0.05),甲氨蝶呤组和健脾解毒汤各组间比较差异均无统计学意义(P均>0.05)。结论健脾解毒汤可能通过促进脾主运化水液功能的恢复来调节脾虚湿盛证银屑病大鼠皮肤AQP3蛋白的表达,恢复皮肤屏障功能。

Objective It is to study the effect of decoction for fortifying spleen and removing toxin on the expression of aquaporins 3(AQP3)in the skin tissues of psoriasis rat with syndrome of spleen deficiency and dampness exuberance,and to explore the mechanism of this decoction in repairing skin barrier function.Methods According to the random number table,72 male SD rats were randomly divided into 6 groups:blank group,model group,methotrexate group,low dose,medium dose and high dose groups of decoction for fortifying spleen and removing toxin,with 12 rats in each group.Except for the rats in the blank group,the rats in the remaining groups were induced with application of imiquimod cream and rhubarb formula granule by gavage to establish the psoriasis models with syndrome of spleen deficiency and dampness exuberance.After modeling,the methotrexate group was given methotrexate tablets 0.97 mg/kg by gavage,and the low,medium and high dose groups of decoction for fortifying spleen and removing toxin were respectively given decoction for fortifying spleen and removing toxin 2.32 g/kg,4.64 g/kg and 9.26 g/kg by gavage,all once daily,continuously treated for 14 days.The phenotypic characteristics of the rats in each group were observed,the histopathological morphology of the skin was observed by HE staining,and the expression of AQP3 protein in the skin tissue was detected by Western blot.Results At the end of gavage,the rats in the model group ate less,defecated more frequently and had dilute feces;the rats in all groups of decoction for fortifying spleen and removing toxin ate significantly better,defecated less frequently and had moderate texture of feces.HE staining showed that the rats in the model group had hyperplastic hypertrophy of the epidermis,with significant parakeratosis with hyperkeratosis,prolonged epidermal protrusions,some epidermal intercellular edema,more neutrophil infiltration in the epidermis and the stratum corneum,dilated blood vessels in the superficial dermis,and scattered or focal infiltration of chronic inflammatory cells;the skin structure of the rats in each group of decoction for fortifying spleen and removing toxin was significantly improved compared with the model group.The expression of AQP3 protein in the skin tissues of rats in the model group was significantly lower than that in the blank group(P<0.05),and the expressions of AQP3 protein in the methotrexate group and each group of decoction for fortifying spleen and removing toxin were significantly higher than that in the model group(all P<0.05),the differences among the methotrexate group and each group of decoction for fortifying spleen and removing toxin were not statistically significant(all P>0.05).Conclusion Decoction for fortifying spleen and removing toxin can regulate the expression of AQP3 protein in the skin of psoriasis rat with syndrome of spleen deficiency and dampness exuberance to restore the skin barrier function maybe by promoting the recovery of function of the spleen in governing transformation and transportation of water and fluid.