何首乌提取物基于IL-23/IL-17轴对银屑病小鼠皮损及抑炎机制研究

Amelioration and Anti-inflammatory Effects of Polygonum Multiflorum Extract on Psoriasis Mice Based on Interleukin-23/Interleukin-17 Axis

目的 探讨何首乌提取物(PME)对银屑病小鼠皮损的改善及抑炎作用,并阐明其可能的作用机制。方法 雌性BALB/c小鼠随机分为正常组、模型组和PME低、中、高剂量组,每组各15只,除正常组外,剩余各组利用咪喹莫特(IMQ)诱导银屑病模型后给药,连续给药7 d。记录各组小鼠背部皮损情况并进行银屑病皮损面积及严重程度指数(PASI)评分;小鼠背部表皮组织行HE染色;免疫组化法检测CD4及K17的阳性表达;酶联免疫吸附测定(ELISA)法检测皮肤组织内肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)表达;实时荧光定量聚合酶链反应(RT-qPCR)检测IL-17A、IL-17F、IL-22、IL-23信使RNA(mRNA)相对表达水平;蛋白质印迹(Western blotting)法检测细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)蛋白相对表达量。结果 与正常组比较,模型组小鼠PASI评分升高(P<0.05);棘层增厚呈嵴状延长,有明显的细胞炎性水肿,皮肤表皮层厚度增大(P<0.05);CD4、K17阳性表达、TNF-α及IL-8含量、IL-17A、IL-17F、IL-22、IL-23 mRNA相对表达水平及ICAM-1、VCAM-1蛋白相对表达量均提高(P<0.05);与模型组比较,PME低、中、高剂量组小鼠银屑病PASI评分降低,炎性细胞浸润减少,棘层畸变改善,表皮层厚度降低(P<0.05);CD4、K17阳性表达、TNF-α及IL-8含量、IL-17A、IL-17F、IL-22和IL-23 mRNA相对表达水平及ICAM-1和VCAM-1蛋白相对表达量降低(P<0.05);PME作用效果呈剂量依赖性。结论 PME可改善IMQ诱导的银屑病小鼠皮损情况,可能与抑制IL-23/IL-17炎性轴有关。

Objective In order to investigate the amelioration and anti-inflammatory effects of polygonum multiflorum extract(PME)on skin lesions of psoriasis mice,and to elucidate its possible mechanism.Methods Female BALB/c mice were randomly divided into normal group,model group,PME low-dose,medium-dose and high-dose groups,with 15 mice in each group.In addition to the normal group,imiquimod(IMQ)was used to induce psoriasis model for seven days.The skin lesions on the back of mice in each group were recorded,the lesion area and psoriasis severity index(PASI)score was performed.HE staining was performed on mouse dorsal epidermis.The positive expression of CD4 and K17 was detected by immunohistochemistry.The expression of tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)in skin tissues was detected by enzyme-linked immunosorbent assay(ELISA).The messenger RNA(mRNA)relative expression levels of IL-17A,IL-17F,IL-22 and IL-23 were detected by quantitative reverse transcription polymerase chain reaction(RT-qPCR).The relative expression levels of intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)proteins were detected by Western blotting.Results Compared with normal group,PASI score of model group was increased(P<0.05).The thickening of the spinous layer was crista-like extension,there was obvious inflammatory edema,and the thickness of the skin epidermis increased(P<0.05).The positive expression of CD4 and K17,the content of IL-8 and TNF-α,the relative expression levels of IL-17A,IL-17F,IL-22 and IL-23 mRNA as well as the relative expression levels of ICAM-1 and VCAM-1 protein were significantly increased (P<0.05). Compared with model group,PASI score,inflammatory cell infiltration,spinous layer distortion and epidermal layer thickness were decreased in low,medium and high dose PME groups(P<0.05). CD4 and K17 positive expression,TNF-α and IL-8 content,IL-17A,IL-17F,IL-22,IL-23 mRNA relative expression level and ICAM-1,VCAM-1 protein relative expression level decreased(P<0.05). The effects of PME were dose-dependent. Conclusion PME could improve the skin lesions of IMQ-induced psoriasis mice,which may be related to the inhibition of IL-23/IL-17 inflammatory axis.