颠倒散对玫瑰痤疮小鼠炎症反应及IL-6/STAT3/NF-κB通路的影响

Effect of Diandaosan on inflammatory response and IL-6/STAT3/NF-κB pathway of mice with rose acne

目的探讨颠倒散(DDS)对玫瑰痤疮小鼠炎症反应、白细胞介素-6(IL-6)/信号转导子及转录激活因子3(STAT3)/核转录因子κB(NF-κB)通路的影响。方法皮内注射LL37建立玫瑰痤疮小鼠模型,随机分为模型组、DDS低剂量(DDS-L,0.5 g/mL DDS)、中剂量(DDS-M,1 g/mL DDS)、高剂量(DDS-H,2 g/mL DDS)组及DDS-H+重组IL-6蛋白(rIL-6)组(2 g/mL DDS+0.1 mg/kg rIL-6),并以皮内注射相同部位生理盐水的小鼠为对照组,每天1次,干预7 d;干预结束后,对小鼠注射部位皮肤红斑进行评分及面积测定;收集皮损组织,ELISA检测IL-6、干扰素-γ(IFN-γ)及肿瘤坏死因子-α(TNF-α),HE染色观察皮损组织病理学变化,甲苯胺蓝检测肥大细胞浸润,免疫组化检测CD4 T阳性细胞数,Western blot检测IL-6/STAT3/NF-κB通路相关蛋白表达。结果与对照组比较,模型组小鼠注射部位皮肤组织损伤严重,皮肤红斑评分及红斑面积增加,皮损组织中IL-6、IFN-γ及TNF-α水平增高,肥大细胞浸润数、CD4 T阳性细胞数增加,IL-6、STAT3及NF-κB蛋白表达显著增加(P均<0.05);与模型组比较,DDS-L组、DDS-M组、DDS-H组病理损伤不同程度减轻,皮肤红斑评分及红斑面积减小,皮损组织中IL-6、IFN-γ及TNF-α水平降低,肥大细胞浸润数、CD4 T阳性细胞数减少,IL-6、STAT3及NF-κB蛋白表达显著降低(P均<0.05),不同剂量DDS组间比较,差异有统计学意义(P均<0.05);rIL-6逆转了对DDS-H对玫瑰痤疮小鼠的保护作用。结论DDS减少玫瑰痤疮小鼠的炎症反应,可能与抑制IL-6/STAT3/NF-κB通路有关。

Objective To investigate the effect of Diandaosan(DDS)on the inflammatory response and interleukin-6(IL-6)/interleukin-6/signal transducer and activator of transcription 3(STAT3)/nuclear factor-κB(NF-κB)pathway of mice with rose acne.Methods The mice models with rose acne were established by intradermal injection of LL37,and were randomly divided into the model group,the DDS-L group(low-dose,0.5 g/mL DDS),the DDS-M group(medium-dose,1 g/mL DDS),the high-dose DDS-H group(2 g/mL DDS)group,and the DDS-H+rIL-6 group(2 g/mL DDS+0.1 mg/kg recombinant IL-6 protein).The mice injected with physiological saline at the same site were used as the control group,with intervention once a day for 7 days.After the intervention,the skin erythema at the injection site of mice was scored and its area was measured.The skin lesion tissues were collected.ELISA detected IL-6,interferon-γ(IFN-γ),and tumor necrosis factor(TNF-α).The histopathological changes were observed by HE staining.The infiltration of mast cells was detected by Toluidine blue.The number of CD4 T positive cells was detected by immunohistochemistry.Western blot was applied to detect the expression of IL-6/STAT3/NF-κB pathway related proteins.Results Compared with the control group,the skin tissues in the model group were seriously injured at the injection site,the skin erythema score and erythema area,the levels of IL-6,IFN-γ,and TNF-α,the number of mast cell infiltration and CD4 T-positive cells all increased.The expression of IL-6,STAT3 and NF-κB protein also significantly increased(all P<0.05).Compared with the model group,the pathological lesions in DDS-L,DDS-M and DDS-H groups were alleviated to different degrees,the skin erythema score,and erythema area,the levels of IL-6,IFN-γand TNF-αin skin lesions,and the number of mast cell infiltration and CD4 T-positive cells all decreased.The expressions of IL-6,STAT3,and NF-κB protein significantly decreased(all P<0.05),and the differences among different doses of DDS were statistically significant(all P<0.05);rIL-6 reversed the protective effect of DDS-H on mice with rose acne.Conclusion DDS reduces the inflammatory response in mice with rose acne,which may be related to the inhibition of the IL-6/STAT3/NF-κB pathway.