解毒祛瘀滋肾方对MRL/lpr狼疮小鼠皮肤组织JAK1/STAT3信号通路的影响

Effect of Jiedu Quyu Zishen Prescription on JAK1/STAT3 Signaling Pathway in Skin Tissue of MRL/lpr Lupus Mice

[目的]探讨解毒祛瘀滋肾方对MRL/lpr狼疮小鼠皮肤组织酪氨酸蛋白激酶1/信号转导与转录激活因子3(Janus kinase 1/signal transducers and activators of transcription 3,JAK1/STAT3)信号通路的影响。[方法]将30只MRL/lpr小鼠随机分成模型组、西药组(强的松组)和中药组(解毒祛瘀滋肾方组),另选10只C57BL/6小鼠作为对照组。中药组予0.5mg·mL^-1的解毒祛瘀滋肾方水煎剂灌胃,西药组予0.5mg·mL^-1的强的松片混悬液灌胃,模型组和对照组予相同体积的0.9%氯化钠溶液灌胃,各组均灌胃1次/d,连续干预8周。干预结束后观察小鼠皮毛、活动等一般性状及浅表淋巴结大小,并采集小鼠血液、皮肤和肾脏等样本。以苏木精-伊红(hematoxylin-eosin,HE)染色法观察皮损组织病理变化,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清JAK1/STAT3通路上游分子白介素-6(interleukin-6,IL-6)、IL-10含量;Western blot检测皮肤组织JAK/STAT信号通路中JAK1、STAT3蛋白表达。[结果]与对照组比较,模型组小鼠皮肤组织病理显示损伤明显,血清IL-6、IL-10含量均明显升高(P<0.01),JAK1、STAT3蛋白表达均明显升高(P<0.01)。与模型组比较,中药组、西药组小鼠皮肤病理损伤明显改善,血清IL-6、IL-10含量明显降低(P<0.01),皮肤JAK1、STAT3蛋白表达明显降低(P<0.01)。与西药组比较,中药组血清IL-6含量差异无统计学意义(P>0.05),而血清IL-10含量明显升高(P<0.05),JAK1、STAT3蛋白表达差异无统计学意义(P>0.05)。[结论]解毒祛瘀滋肾方能够降低MRL/lpr小鼠血清IL-6、IL-10含量,可抑制皮肤组织中JAK1、STAT3蛋白表达,提示阻断JAK/STAT信号通路可能是解毒祛瘀滋肾方治疗系统性红斑狼疮的作用机制之一。

[Objective]To investigate the effect of Jiedu Quyu Zishen Prescription on Janus kinase 1/signal transducers and activators of transcription 3(JAK1/STAT3)signaling pathway in skin tissue of MRL/lpr lupus mice.[Methods]Thirty MRL/lpr mice were randomly divided into model group,western medicine group and traditional Chinese medicine(TCM)group,and another ten C57BL/6 mice were selected as control group.Mice in TCM group were given 0.5mg·mL^-1 of Jiedu Quyu Zishen Prescription,while western medicine group was given 0.5mg·mL^-1 of prednisone acetate suspension,model group and control group were given the same volume of distilled water,once a day,for 8 weeks.After the intervention,the skin,activity and superficial lymph nodes of mice were observed,and blood,skin and kidney samples were collected.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of skin lesions,the serum contents of interleukin-6(IL-6)and IL-10 in the upper reaches of serum JAK1/STAT3 signaling pathway were detected by enzyme linked immunosorbent assay(ELISA),and the protein expression of JAK1 and STAT3 in skin JAK/STAT signaling pathway was detected by Western blot.[Results]Compared with control group,the pathological changes of skin tissue in model group showed obvious damage,and the serum contents of IL-6 and IL-10 and the protein expression of JAK1 and STAT3 significantly increased in model group(P<0.01).Compared with model group,the skin pathological damage of the mice in TCM group and western medicine group was significantly improved,the serum contents of IL-6 and IL-10 and the protein expression of JAK1 and STAT3 significantly decreased(P<0.01).Compared with western medicine group,there was no significant difference in serum content of IL-6 in TCM group(P>0.05),while the serum content of IL-10 significantly increased(P<0.05),and there was no significant difference in protein expression of JAK1 and STAT3(P>0.05).[Conclusion]Jiedu Quyu Zishen Prescription can reduce the serum contents of IL-6 and IL-10 of MRL/lpr mice,and decrease the protein expression of JAK1 and STAT3 in skin tissue,which suggests that blocking of JAK/STAT signaling pathway may be one of the mechanisms of Jiedu Quyu Zishen Prescription in the treatment of systemic lupus erythematosus.