摘要
目的:探讨复方生地合剂对MRL/lpr小鼠Janus激酶(JAK)/信号转导和转录激活因子(STAT)信号通路的影响。方法:将30只MRL/lpr小鼠随机分组为模型组、复方生地合剂组、强的松组,每组10只;取10只雌性C57BL/6小鼠作为空白组。复方生地合剂组小鼠予复方生地合剂药液灌胃(27.7 g/kg),强的松组小鼠予强的松片混悬液灌胃(12.3 mg/kg),模型组、空白组小鼠予0.9%氯化钠溶液灌胃(0.3 mL/次),各组均每日给药1次,灌胃8周。观察各组小鼠一般情况、死亡情况、脾脏指数;采用酶联免疫吸附试验(ELISA)法检测血清白细胞介素-2(IL-2)、白细胞介素-17(IL-17)、抗ds-DNA、免疫球蛋白G(IgG)水平;采用Western blotting检测小鼠Janus激酶2(JAK2)、磷酸化Janus激酶2(p-JAK2)、信号转导和转录激活因子1(STAT1)和磷酸化信号转导和转录激活因子1(p-STAT1)蛋白表达;采用HE染色观察肾组织病理变化;采用免疫组化染色检测织p-JAK2阳性表达。结果:模型组、复方生地合剂组、强的松组各有1只小鼠死亡,空白组无小鼠死亡。模型组小鼠脾脏指数高于空白组(P<0.05);复方生地合剂组、强的松组小鼠脾脏指数均低于模型组(P<0.05)。模型组小鼠血清IL-17、IgG、抗ds-DNA水平高于空白组,IL-2水平低于空白组(P<0.05);复方生地合剂组、强的松组小鼠血清IL-17、IgG、ds-DNA水平均低于模型组(P<0.05),IL-2水平高于模型组(P<0.05)。模型组小鼠肾组织JAK2、p-JAK2、STAT1及p-STAT1蛋白相对表达量高于空白组(P<0.05);复方生地合剂组、强的松组小鼠肾组织JAK2、p-JAK2、STAT1及p-STAT1蛋白相对表达量均低于模型组(P<0.05)。模型组小鼠肾组织p-JAK2阳性表达水平高于空白组(P<0.05);复方生地合剂组、强的松组小鼠肾组织p-JAK2阳性表达水平均低于模型组(P<0.05)。模型组肾脏病理显示系膜细胞及内皮细胞增生,大范围炎症细胞浸润;复方生地合剂组、强的松组可见少量系膜细胞及内皮细胞增生,轻微炎症细胞浸润,轻度肾小管变性。结论:复方生地合剂可能通过抑制JAK2/STAT1信号通路,抑制MRL/lpr小鼠相关炎症因子和抗体表达。
Objective:To explore effect of compound Shengdi mixture(CSM)on janus kinase(JAK)/signal transduction and activator of transcription(STAT)in mice of systemic lupus erythematosus.Methods:Totally 30 MRL/lpr mice were randomly divided into model group,CSM group and prednisone group,with 10 mice in each group.And 10 female C57BL/6 mice were used as blank group.The mice were given CSM liquid medicine(27.7 g/kg)by gavage in CSM group,and prednisone tablet suspension(12.3 mg/kg)by gavage in prednisone group.The mice were given 0.9%sodium chloride solution by gavage(0.3 mL per time)in the model group and blank group,once a day for 8 weeks.The general situation,death and spleen index were observed.The levels of interleukin-2(IL-2),interleukin-17(IL-17),anti-ds-DNA and immunoglobulin G(IgG)in serum were detected by enzyme-linked immunosorbent assay(ELISA).Western blotting was used to detect the expression of janus kinase 2(JAK2),phosphorylase JAK2(p-JAK2),signal transduction and activator of transcription 1(STAT1)and phosphorylase STAT1(p-STAT1)proteins in mice.HE staining was used to observe the pathological changes of kidney tissue.Immunohistochemical staining was used to detect the positive expression of p-JAK2.Results:There was one mouse died in the model group,CSM group and prednisone group,and there was no mouse died in blank group.The model group showed higher spleen index than blank group(P<0.05).The CSM group and prednisone group showed lower spleen index than model group(P<0.05).The model group showed higher levels of serum IL-17,IgG and anti-ds-DNA than blank group,while lower level of IL-2 than blank group(P<0.05).The CSM group and prednisone group showed lower levels of serum IL-17,IgG and anti-ds-DNA than model group,while higher level of IL-2 than model group(P<0.05).The model group showed higher protein contents of JAK2,p-JAK2,STAT1 and p-STAT1 in renal tissue than blank group(P<0.05).The CSM group and prednisone group showed lower protein contents of JAK2,p-JAK2,STAT1 and p-STAT1 in renal tissue than model group(P<0.05).The model group showed higher positive expression of p-JAK2 in kidney than blank group(P<0.05).The CSM group and prednisone group showed lower positive expression of p-JAK2 in kidney than model group(P<0.05).Renal pathology in model group showed that mesangial cells and endothelial cells proliferated and inflammatory cells infiltrated in a large range.In CSM group and prednisone group,a small amount of mesangial cells and endothelial cells proliferated,with slight inflammatory infiltration and mild renal tubular degeneration.Conclusion:Compound Shengdi mixture may inhibit the expression of inflammatory factors and antibodies through inhibition of JAK2/STAT1 pathway.
作者
朱博玉
陈薇薇
刘智超
常靖升
黄慧萍
ZHU Boyu;CHEN Weiwei;LIU Zhichao;CHANG Jingsheng;HUANG Huiping(Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China)
出处
《中医药导报》
2025年第8期41-46,共6页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
上海医学创新发展基金会中医药科技发展项目(WL-YJXYS-2021005K)
上海医学创新发展基金会中医药传承发展项目(WLJH2021ZY-MZY037)
上海市医院中药制剂产业转化协同创新中心项目(K211009N)。
