摘要
目的:探讨加味地榆清疕膏对咪喹莫特乳膏(imiquimod cream,IMQ)诱导的银屑病模型小鼠的治疗作用及其机制。方法:将50只BALB/c小鼠随机分为5组:空白组、模型组、加味地榆清疕膏组(加味组)、地榆清疕膏组(地榆组)、卡泊三醇软膏组(卡泊组),每组10只。空白组小鼠每日仅涂抹凡士林50 mg。其余4组每日均涂抹IMQ 50 mg以造模,6 h后分别涂抹:模型组涂抹凡士林50 mg,加味组、地榆组、卡泊组分别涂抹相应药膏50 mg,连续7 d。第7天进行银屑病皮损面积及严重性指数(psoriasis area and severity index,PASI)评分;干预结束后处死小鼠,取小鼠银屑病皮损进行HE染色并观察其病理形态学变化,ELISA法检测小鼠血清白细胞介素6(interleukin-6,IL-6)、白细胞介素22(interleukin-22,IL-22)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的表达水平,qPCR法检测皮损组织中酪氨酸激酶1(janus kinase 1,JAK1)、信号转导与转录激活因子3(signal transducer and activator of transcription 3,STAT3)的mRNA表达水平,Western blot法检测皮损组织中JAK1、p-STAT3/STAT3、Survivin蛋白的表达水平。结果:HE染色结果表明,模型组小鼠出现红斑、鳞屑、浸润性皮损及角化不全、角化过度、Munro微脓肿等银屑病样病理变化;与模型组相比,各治疗组病理表现均有不同程度减轻且PASI评分明显降低,其中加味组PASI评分低于卡泊组。ELISA检测结果表明,与空白组相比,模型组IL-6、IL-22、TNF-α水平明显升高(P<0.05);与模型组相比,各治疗组IL-6、IL-22、TNF-α水平明显降低(P<0.05);加味组、地榆组IL-6水平明显低于卡泊组(P<0.05),加味组IL-22水平明显低于地榆组、卡泊组(P<0.05)。qPCR和Western blot结果表明,与空白组相比,模型组JAK1、p-STAT3/STAT3和Survivin蛋白表达水平、JAK1 mRNA、STAT3 mRNA表达水平均明显升高(P<0.05);与模型组相比,各治疗组JAK1、p-STAT3/STAT3和Survivin蛋白表达水平、JAK1 mRNA、STAT3 mRNA表达水平均明显降低(P<0.05);加味组p-STAT3/STAT3明显低于地榆组(P<0.05),加味组JAK1 mRNA表达水平明显低于卡泊组(P<0.05)。结论:加味地榆清疕膏可以减轻银屑病模型小鼠皮损的严重程度并改善其病理表现,整体优于地榆清疕膏,其作用机制可能与降低IL-6、IL-22、TNF-α等炎性细胞因子的表达、抑制JAK1/STAT3信号通路及凋亡抑制蛋白Survivin的表达有关。
Objective:To investigate the therapeutic effects and mechanism of Diyu Qingbi Ointment on imiquimod cream(IMQ)-induced psoriasis model mice.Methods:Fifty BALB/c mice were randomly divided into five groups:blank group,model group,modified Jiawei Qingbi group(treated with modified Diyu Qingbi Ointment),Diyu group(treated with Diyu Qingbi Ointment),and Calcipotriol ointment group.Each group contained 10 mice.The blank group received daily application of Vaseline ointment,while the remaining groups were subjected to daily 5%IMQ application to induce the model,followed 6 hours later by the corresponding drug intervention,continuously for 7 days.On day 7,psoriasis lesion area and severity index(PASI)scores were recorded;after the intervention,mice were sacrificed,psoriatic lesions were collected for HE staining to observe pathological morphological changes.ELISA was used to detect serum expression levels of interleukin-6(IL-6),interleukin-22(IL-22),and tumor necrosis factor-alpha(TNF-α).qPCR was used to detect mRNA expression levels of Janus kinase 1(JAK1)and signal transducer and activator of transcription 3(STAT3)in lesion tissues;Western blot analysis was used to detect the expression levels of JAK1,p-STAT3/STAT3,and Survivin proteins in psoriatic lesions.Results:HE staining results showed that mice in the model group exhibited erythema,scaling,infiltrative lesions,as well as abnormal keratinization,hyperkeratosis,and Munro microabscesses,indicating psoriasis-like pathological changes.Compared with the model group,the pathological manifestations of each treatment group were alleviated to varying degrees,and PASI scores were significantly reduced,with the modified formula group showing lower PASI scores than the calcipotriol group(P<0.05).ELISA results indicated that compared with the blank group,the model group had significantly elevated levels of IL-6,IL-22,and TNF-α(P<0.05);compared with the model group,levels of IL-6,IL-22,and TNF-αin each treatment group were significantly reduced(P<0.05);IL-6 levels in the modified formula and Sanguisorba officinalis groups were significantly lower than those in the calcipotriol group(P<0.05),and IL-22 levels in the modified formula group were significantly lower than those in both the Sanguisorba officinalis and calcipotriol groups(P<0.05).qPCR and Western blot results showed that compared with the blank group,the model group had significantly higher protein expression levels of JAK1,p-STAT3/STAT3,and Survivin,as well as elevated mRNA levels of JAK1 and STAT3(P<0.05);compared with the model group,expression levels of JAK1,p-STAT3/STAT3,Survivin proteins,and JAK1 and STAT3 mRNA were significantly reduced in the treatment groups(P<0.05);p-STAT3/STAT3 in the modified formula group was significantly lower than in the Sanguisorba officinalis group(P<0.05),and JAK1 mRNA expression in the modified formula group was significantly lower than in the calcipotriol group(P<0.05).Conclusion:Modified Diyu Qingbi Ointment can reduce the severity of skin lesions in psoriatic model mice and improve their pathological manifestations,performing overall better than Sanguisorba officinalis Qingbi ointment.Its mechanism may be related to reducing the expression of inflammatory cytokines such as IL-6,IL-22,TNF-α,and inhibiting the JAK1/STAT3 signaling pathway as well as the expression of the apoptosis-inhibiting protein Survivin.
作者
王凯利
段行武
陈曦
朱泽兵
吴治民
WANG Kaili;DUAN Xingwu;CHEN Xi;ZHU Zebing;WU Zhimin(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing China 100700;Hubei Provincial Hospital of Chinese Medicine,Wuhan Hubei China 430073;The Third Affiliated Hospital to Guangzhou University of Chinese Medicine(Guangdong Provincial Institute of Traditional Chinese Medicine Orthopedics),Guangzhou Guangdong China 510378)
出处
《中医学报》
2025年第12期2661-2667,共7页
Acta Chinese Medicine
基金
国家自然科学基金项目(82074436)。
