1.广州中医药大学东莞医院,广东省东莞市东城街道松山湖大道东城段3号,523000
2.广州中医药大学第二临床医学院
3.广州中医药大学第二附属医院省部共建中医湿证国家重点实验室
4.中医药广东省实验室(横琴实验室)
5.广东省中医药科学院
linghan99@gzucm.edu.cn
收稿:2025-01-02,
修回:2025-04-22,
纸质出版:2025-07-02
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张雅婷,苏浩杰,刘凡露等.外湿因素诱导下银屑病脾虚湿阻证病证结合大鼠模型的建立[J].中医杂志,2025,66(13):1369-1377.
ZHANG Yating,SU Haojie,LIU Fanlu,et al.Establishment of Psoriasis Rat Model with Spleen Deficiency and Dampness Obstruction Syndrome Induced by External Dampness Factors[J].Journal of Traditional Chinese Medicine,2025,66(13):1369-1377.
张雅婷,苏浩杰,刘凡露等.外湿因素诱导下银屑病脾虚湿阻证病证结合大鼠模型的建立[J].中医杂志,2025,66(13):1369-1377. DOI: 10.13288/j.11-2166/r.2025.13.012.
ZHANG Yating,SU Haojie,LIU Fanlu,et al.Establishment of Psoriasis Rat Model with Spleen Deficiency and Dampness Obstruction Syndrome Induced by External Dampness Factors[J].Journal of Traditional Chinese Medicine,2025,66(13):1369-1377. DOI: 10.13288/j.11-2166/r.2025.13.012.
目的
2
构建外湿型银屑病脾虚湿阻证病证动物模型并对其进行评价。
方法
2
22只SD大鼠随机分为正常对照组3只、寻常型银屑病组5只、外湿型脾虚湿阻证组7只、外湿型银屑病脾虚湿阻证组7只。采用人工模拟高湿环境32周建立外湿型脾虚湿阻证大鼠模型,外涂咪喹莫特乳膏7天建立寻常型银屑病大鼠模型,并二者结合构建外湿型银屑病脾虚湿阻证大鼠模型。正常组大鼠饲养于正常湿度环境。观察大鼠整体状态、舌象变化评价大鼠宏观证候表现;通过脂肪组织和肝脏组织变化评价大鼠微观证候表现;通过皮肤病理变化、银屑病面积和严重程度指数(PASI)、增殖细胞核抗原(PCNA)表达和脾脏组织变化评价大鼠银屑病严重程度;通过流式细胞术检测大鼠CD4
+
干扰素-γ
+
细胞(CD4
+
IFN-γ
+
细胞)、CD4
+
肿瘤坏死因子-α
+
细胞(CD4
+
TNF-α
+
细胞)和叉头框蛋白P3
+
调节性T细胞(CD3
+
CD4
+
FoxP3
+
Treg细胞)变化。
结果
2
在宏观表现上,外湿型脾虚湿阻证组大鼠及外湿型银屑病脾虚湿阻证组大鼠均出现精神萎靡、扎堆嗜睡、毛发枯黄且杂乱无光等脾虚湿阻证表现,部分可见软便稀便、肛周污秽;外湿型脾虚湿阻证组及外湿型银屑病脾虚湿阻证组大鼠均可见舌体胖大,舌质淡而胖润,舌面水滑。在微观表现上,与寻常型银屑病组比较,外湿型银屑病脾虚湿阻证组附睾脂肪指数上升(
P
<0.05);与正常对照组和外湿型脾虚湿阻证组大鼠比较,外湿型银屑病脾虚湿阻证组大鼠脾脏质量显著上升(
P
<0.05);各组大鼠肝脏表观及HE染色表明肝脏组织无明显病变。大鼠背部皮损表观可见外湿型银屑病脾虚湿阻证组大鼠背部皮损情况较寻常型银屑病组大鼠明显加重。与正常对照组及外湿型脾虚湿阻证组比较,寻常型银屑病组和外湿型银屑病脾虚湿阻证组大鼠红斑评分、鳞屑评分、浸润评分、PASI总分以及CD3
+
CD4
+
FoxP3
+
Treg细胞比例均显著升高(
P
<0.05),皮肤表皮基底层及真皮层均可见PCNA阳性表达明显增多;与外湿型脾虚湿阻证组比较,外湿型银屑病脾虚湿阻证组CD4
+
TNF-α
+
细胞比例显著上升(
P
<0.05);各组间CD4
+
IFN-γ
+
细胞比例无显著差异(
P
>0.05)。
结论
2
采用人工模拟高湿环境结合咪喹莫特诱导可成功构建银屑病外湿型银屑病脾虚湿阻证病证结合大鼠模型。
Objective
2
To construct a rat model of psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type), and evaluate the macroscopic manifestations and microscopic indicators of the model.
Methods
2
Twenty-two SD rats were divided into normal group (n=3), common psoriasis group (n=5), spleen deficiency and dampness obstruction syndrome (external dampness type) group (n=7), and psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group (n=7). The spleen deficiency and dampness obstruction syndrome (external dampness type) rat model was established through 32-week exposure to an artificially simulated high-humidity environment, while the common psoriasis model was developed via 7-day topical application of imiquimod cream, and these two approaches were combined to construct a composite model of psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type). Rats in the normal group were housed under normal humidity conditions. The general state, tongue manifestation of rats were observed to evaluate the macroscopic syndrome manifestations; the microscopic syndrome manifestations of rats were evaluated through adipose tissue and liver tissue changes; the severity of psoriasis in rats was evaluated through skin pathological changes, psoriasis area and severity index (PASI), proliferating cell nuclear antigen (PCNA) expression and spleen tissue changes; changes in rat CD4
+
interferon-γ
+
cells (CD4
+
IFN-γ
+
cells), CD4
+
tumour necrosis factor-α+ cells (CD4
+
TNF-α
+
cells), and forkhead framing protein P3
+
regulatory T cells (CD3
+
CD4
+
FoxP3
+
Treg cells) were detected by flow cytometry.
Results
2
Macroscopically, both the spleen deficiency and dampness obstruction syndrome (external dampness type) group and psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group exhibited manifestati
ons of spleen deficiency and dampness obstruction, including lethargy, huddling behavior, dull and disheveled fur, as well as soft or loose stools and perianal soiling in some individuals; both these two groups displayed enlarged tongue, swollen, and moist tongue texture, accompanied by slippery tongue surface. Microscopically, compared to the common psoriasis group, the psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group showed increased epididymal fat index (
P
<0.05); compared to the normal group and spleen deficiency and dampness obstruction syndrome (external dampness type) group, the psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group demonstrated significantly elevated spleen mass (
P
<0.05), while hepatic gross morphology and HE staining revealed no significant histopathological changes across all groups. Dorsal skin lesions were markedly exacerbated in the psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group when compared to those in common psoriasis group. Both the common psoriasis group and psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group exhibited significantly higher erythema scores, scaling scores, infiltration scores, PASI total scores, and proportions of CD3
+
CD4
+
FoxP3
+
Treg cells compared to the normal group and spleen deficiency and dampness obstruction syndrome (external dampness type) group (
P
<0.05), with pronounced PCNA-positive expression observed in the epidermal basal layer and dermis; the psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) group displayed significantly increased proportions of CD4
+
TNF-α
+
cells compared to the spleen deficiency and dampness obstruction syndrome (external dampness type) group (
P
<0.05); whereas no significant differences w
ere detected in CD4
+
IFN-γ
+
cell proportions among groups (
P
>0.05).
Conclusion
2
The rat model of psoriasis with spleen deficiency and dampness obstruction syndrome (external dampness type) can be successfully constructed by artificially simulating a high-humidity environment combined with imiquimod induction.
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