夏枯草水提液对实验性自身免疫性甲状腺炎的治疗作用及机制研究  

Effect of aqueous extract of Prunella vulgaris on experimental autoimmune thyroiditis in rats

作　　者：傅晓丹[1] 章晓芳[1] 马丽珍[1] 卓广超[2] 陈岳明[3] 王炜[4] 张险峰[1] 张楚[1] 俞灵莺[1] FU Xiaodan;ZHANG Xiaofang;MA Lizhen(Department of Endocrinology,Hangzhou First People’s Hospital,Hangzhou 310006,China)

机构地区：[1]杭州市第一人民医院内分泌科,310006 [2]杭州市第一人民医院中心试验室,310006 [3]杭州市第一人民医院基因实验室,310006 [4]杭州市第一人民医院病理实验室,310006

出　　处：《浙江医学》2020年第9期893-897,I0004,共6页Zhejiang Medical Journal

基　　金：浙江省中医药科学研究计划(2013ZA108);杭州市农业与社会发展科研自主申报项目(20191203B83)。

摘　　要：目的探讨夏枯草水提液(PVAE)对实验性自身免疫甲状腺炎(EAT)大鼠的治疗作用及机制。方法用猪甲状腺球蛋白和完全弗氏佐剂制作EAT大鼠模型,随机分为PVAE预防性治疗组(P组)、PVAE低剂量治疗组(TL组)、PVAE高剂量治疗组(TH组)、EAT模型对照组(M组),并设立正常对照组(C组),每组14只。P组于造模前2周开始予PVAE 10g/(kg·d)灌胃给药,TL组、TH组于造模后2周开始分别予PVAE 10g/(kg·d)、2.5g/(kg·d)灌胃给药,M组和C组于造模后2周开始予去离子水2ml/d灌胃,连续给药6周。实验结束后检测各组大鼠血清抗甲状腺球蛋白抗体(TgAb)、甲状腺过氧化物酶抗体(TPOAb)、磷脂酰肌醇-3-激酶(PI3K)-丝氨酸-苏氨酸蛋白激酶B(AKt)-核转录因子(NF-κB)途经下游产物趋化因子受体3(CXCR3)和趋化因子10(CXCL10)水平;观察甲状腺组织的病理改变,检测CXCR3、CXCL10、Akt和NF-κB的m RNA、蛋白及表达水平。结果M组大鼠血清Tg Ab和TPOAb水平较C组呈明显升高,甲状腺组织有不同程度的滤泡结构破坏和淋巴细胞浸润,提示EAT大鼠造模成功。PVAE各干预组的甲状腺滤泡均较M组完整,间质水肿减轻,少量炎症细胞浸润,其中P组TPOAb水平较M组明显降低(P<0.01)。PVAE各干预组的CXCL10水平均较M组下降,其中P组与C组比较差异无统计学意义(P>0.05)。PVAE治疗组(TH、TL组)的Akt m RNA、CXCR3蛋白表达及TH组的NF-κB m RNA、Akt蛋白表达显著低于M组(P<0.05或0.01)。免疫组化结果显示各组大鼠甲状腺组织Akt均呈阴性表达,NF-κB、CXCR3及CXCL10在甲状腺滤泡细胞的细胞质和细胞核中呈不同程度阳性表达。M组NF-κB表达量明显高于C组(P<0.05),且经PVAE干预后各组NF-κB表达量均呈不同程度下调,其中P组与C组比较差异无统计学意义(P>0.05)。结论PI3K途径参与EAT的形成,PVAE干预可能通过抑制PI3K途径减轻自身免疫性甲状腺炎。Objective To investigate the effects of aqueous extract of Prunella vulgaris(PVAE)on experimental autoimmune thyroiditis(EAT)in rats and its relation with phosphatidylinositol-3-kinase(PI3K)pathway.Methods EAT rats were randomly divided into 5 groups:PVAE pre-treatment group(group P,n=14),PVAE low dose treatment group(group TL,n=14),PVAE high dose treatment group(group TH,n=14),EAT model control group(group M,n=13)and normal control group(group C,n=14).PVAE of 10 g/(kg·d)was given to rats in groups P and TH orally,PVAE of 2.5 g/(kg·d)was given to rats in group TL,while rats in groups M and C were given deionized water at 2 ml/d.Treatment was offered for 6 weeks.Serum levels of autoantibody(TgAb and TPOAb),chemokine CXCL10 and its receptor CXCR3 were determined after the treatment.The pathological changes in thyroid tissues and mRNA expression of serine-threonine protein kinase B(Akt),nuclear factor kappa BNFκB(NF-κB),CXCL10 and CXCR3 were detected by immunohistochemistry and real-time PCR.The protein expressions of phosphorylated Akt,NF-κB and CXCR3 were detected by Western blot.Results Compared to group C,serum TgAb,TPOAb and the pathological changes in thyroid tissue were significantly increased in EAT rats.The serum TPOAb level in group P was decreased and the pathological changes in thyroid tissue of all PVAE groups were attenuated compared with group M(P<0.01).The level of serum CXCL10 was reduced in group TH,TL and P,and it showed no difference in groups M and C.The expression of Akt mRNA and CXCR3 protein were reduced in groups TH and TL(both P<0.05,respectively),and the levels of Akt protein and NF-κB mRNA also reduced in group TH(both P<0.01,respectively).The immunohistochemical staining of Akt was negative in thyroid follicular cells(TFCs)or in lymphocytes in all groups.The expression of NF-κB,CXCR3 and CXCL10 in TFCs were positive in both cytoplasm and nuclear;compared with group C,the expression of NFκBNF-κB was siginificantly increased in group M(P<0.05).When treated with PVAE,the expression

关 键 词：夏枯草 实验性自身免疫甲状腺炎 趋化因子 磷脂酰肌醇-3-激酶 

分 类 号：R285.5[医药卫生—中药学]