摘要
目的:观察槐杞黄颗粒调节Th17/Treg平衡对银屑病样小鼠皮损的干预作用,揭示其调节免疫发挥抗炎作用及改善皮损延缓复发的效应。方法:选取48只雄性KM小鼠,随机分为空白组、模型组、阳性组、高剂量组、中剂量组、低剂量组,每组各8只,采用背部涂抹5%咪喹莫特乳膏(IMQ)诱导建立银屑病小鼠模型,空白组涂抹等量凡士林对照,边造模边灌胃给药,共7 d,末次给药1 h后于小鼠右耳廓涂抹20μl二甲苯建立耳肿胀模型,观察小鼠耳肿胀程度,计算各组小鼠脾脏指数及体重变化;实验期间对小鼠背部皮损进行严重程度指数(PASI)评分。苏木精-伊红(HE)染色观察背部皮损病理学改变,并测量表皮厚度;酶联免疫吸附测定法(ELISA)检测小鼠血清白细胞介素(IL)-17、IL-6、IL-10、转化生长因子(TGF-β)水平;实时荧光定量PCR(RT-qPCR)检测小鼠脾脏组织叉头框蛋白P3(FoxP3)、维甲酸相关孤儿受体γt(RORγt)、转录激活因子3(STAT3)表达。结果:与空白组相比,模型组小鼠背部皮肤增厚,出现红斑、鳞屑等银屑病样改变,PASI评分升高;耳肿胀明显,脾脏指数升高,体重增长明显缓慢(P<0.01);HE染色表现为表皮细胞角化不全、棘层增厚,真皮层炎性细胞浸润,测量表皮厚度明显增厚(P<0.05);RORγt mRNA、STAT3 mRNA表达、炎症因子IL-17与IL-6水平较空白组升高(P<0.01),FoxP3 mRNA表达、IL-10与TGF-β水平降低(P<0.01)。与模型组相比,各治疗组皮损增厚减轻,PASI评分降低,病理学表现改善;耳肿胀得到抑制,脾脏指数降低(P<0.01),体重变化改善(P<0.05),RORγt mRNA、STAT3 mRNA表达、炎症因子IL-17与IL-6水平降低(P<0.01),FoxP3 mRNA表达、IL-10与TGF-β水平升高(P<0.01)。结论:槐杞黄颗粒通过抑制IL-17/IL-6/STAT3/RORγt轴、上调IL-10/TGF-β/FoxP3通路,恢复Th17/Treg平衡,减轻炎症反应及表皮增生,改善其皮损症状。
Objective:To observe the effect of Huaiqihuang granules in regulating the Th17/Treg balance on psoriasiform skin lesions in mice,and to elucidate its immunomodulatory anti-inflammatory effects in improving skin lesions and delaying recurrence.Methods:A total of 48 male KM mice were randomly divided into six groups:blank group,model group,positive control group,high-dose group,medium-dose group,and low-dose group,with 8 mice in each group.A psoriasis mouse model was established by applying 5%imiquimod(IMQ)cream on the dorsal skin.The blank group was treated with an equal amount of Vaseline as control.Drug administration was performed via gavage concurrently during modeling for 7 consecutive days.One hour after the final administration,20μl of xylene was applied to the right auricle of each mouse to establish an ear swelling model.The degree of ear swelling was observed,and spleen index and body weight changes were calculated for each group.During the experiment,the severity of dorsal skin lesions was evaluated using the Psoriasis Area and Severity Index(PASI)score.Hematoxylin-Eosin(HE)staining was used to observe pathological changes in dorsal skin lesions and to measure epidermal thickness.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of interleukin(IL)-17,IL-6,IL-10,and transforming growth factor(TGF-β).Quantitative real-time PCR(qPCR)was employed to detect the expression of forkhead box protein P3(FoxP3),retinoic acid-related orphan receptorγt(RORγt),and signal transducer and activator of transcription 3(STAT3)in spleen tissues.Results:Compared with the blank group,mice in the model group showed dorsal skin thickening,erythema,scaling,and other psoriasiform changes,with increased PASI scores;significant ear swelling,increased spleen index,and markedly slower body weight gain were observed(P<0.01).HE staining revealed parakeratosis,thickening of the stratum spinosum,and inflammatory cell infiltration in the dermis,with significantly increased epidermal thickness(P<0.05).The expression levels of RORγt mRNA and STAT3 mRNA,as well as the levels of inflammatory cytokines IL-17 and IL-6,were significantly elevated compared with the blank group(P<0.01),while the expression of FoxP3 mRNA and the levels of IL-10 and TGF-βwere significantly decreased(P<0.01).Compared with the model group,the treatment groups showed reduced skin lesion thickening,decreased PASI scores,and improved pathological manifestations;ear swelling was suppressed,spleen index was reduced(P<0.01),and body weight changes were improved(P<0.05).The expression levels of RORγt mRNA and STAT3 mRNA,as well as the levels of inflammatory cytokines IL-17 and IL-6,were decreased(P<0.01),while the expression of FoxP3 mRNA and the levels of IL-10 and TGF-βwere increased(P<0.01).Conclusion:Huaiqihuang granules alleviate inflammatory responses and epidermal hyperplasia,and improve skin lesion symptoms by inhibiting the IL-17/IL-6/STAT3/RORγt axis,upregulating the IL-10/TGF-β/FoxP3 pathway,and restoring the Th17/Treg balance.
作者
张雨冰
何海艳
焦滕震
李丹
贾颖
ZHANG Yubing;HE Haiyan;JIAO Tengzhen;LI Dan;JIA Ying(The First Clinical College,Shanxi Univresity of Traditional Chinese Medicine,Jinzhong 030619,China)
出处
《陕西中医》
2025年第10期1318-1324,共7页
Shaanxi Journal of Traditional Chinese Medicine
基金
山西省自然科学基金资助面上项目(202403021211186)
湖北陈孝平科技发展基金资助免疫性疾病研究槐杞黄专项基金资助项目(CXPJJH123004-029)
山西省中医药管理局中医药创新团队建设项目(ZYYTD2024033)。
