Objective:To investigate the effect of Fuzheng Xiaoying Decoction on experimental autoimmune thyroiditis(EAT)model rats and explore its possible mechanism of immune regulation.Methods:Among the 40 female SD rats,10 we...Objective:To investigate the effect of Fuzheng Xiaoying Decoction on experimental autoimmune thyroiditis(EAT)model rats and explore its possible mechanism of immune regulation.Methods:Among the 40 female SD rats,10 were randomly selected as the blank group,and the rest were immunized with pig thyroglobulin and Freund's adjuvant combined with high iodine feeding to make the EAT model.After the model was established,they were randomly divided into three groups:the model group,the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group,which were continuously gavaged for 2 months.The anti thyroid autoantibodies,thyroid function,IL-38 and Th17/Treg distribution were measured.The pathological changes of thyroid tissue were observed by HE staining.Results:Compared with the blank group,the levels of TPOAb and TGAb in the model group were significantly higher(P<0.05).Lymphocyte infiltration was seen in the thyroid tissue of the model group,and also the thyroid follicles were partially destroyed,the shape was irregular,and the colloid distribution was uneven,indicating that the modeling was successful.Compared with the model group,the levels of TPOAb and TGAb in the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group decreased significantly(P<0.05).And the Fuzheng Xiaoying Decoction group had more significant improvement in thyroid follicle destruction and lymphocyte infiltration than the selenium yeast tablet group(P<0.05).Compared with the blank group,the levels of T3,T4,FT3 and FT4 in the model group increased,the level of IL-38 decreased,the proportion of Th17 increased,and the ratio of Th17/Treg increased(P<0.05).Compared with the model group,T4 level decreased,IL-38 level increased in the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group(P<0.05).But there was no statistically significant difference in the reduction of Th17 proportion and Th17/Treg ratio between the above two groups and the model group(P>0.05).There was no significant difference in Treg ratio among the groups.Conclusion:Fuzheng Xiaoying Decoction can significantly reduce the level of anti thyroid autoantibodies in EAT rats by regulating the immunity of EAT rats,and improve the follicular destruction and lymphocyte infiltration of thyroid tissue in rats.展开更多
Objective： To explore the regulatory mechanism of Xiaoyin Recipe （消银方） on the T helper 1/ T helper 2 （Th1/Th2） immune balance. Methods： Thirty-six experimental animals were divided into three groups, 12 rats ...Objective： To explore the regulatory mechanism of Xiaoyin Recipe （消银方） on the T helper 1/ T helper 2 （Th1/Th2） immune balance. Methods： Thirty-six experimental animals were divided into three groups, 12 rats in each group： blank control group （B group）, negative control group （N group）, and Xiaoyin Recipe treatment group （T group）. The latter two groups received immunization of experimental autoimmune thyroiditis （EAT）, and T group were treated with Xiaoyin Recipe for a month. Then, the expression of Th1-Th2-related genes in peripheral blood mononuclear cells （PBMCs） were screened with Oligo GEArray Rat Thl-Th2-Th3 Microarray. The expressions of tumor necrosis factor- α （TNF- α）, interleukin-10 （IL-10）, T-box expressed in T-cells （T-bet）, and GATA-binding protein-3 （GATA-3） were detected by real-time polymerase chain reaction （RT-PCR）. Results： Gene array screening showed that compared to N group, in T group after Xiaoyin Recipe treatment, 3 genes were upregulated in EAT rats, including interleukin-27 receptor alpha （IL-27roL）, glomulin （GImn）, and GATA-3, while 38 genes were downregulated, such as CD28, IL-18, signal transducer, and activator of transcription 1 （STAT1）, T-bet, TNF receptor superfamily member 4 （TNFRSF4）, TNF ligand superfamily member 5 （TNFSF5）, and TNF receptor superfamily member 5 （TNFRSF5）. While RT-PCR showed that there was an increased level of TNF-α mRNA （P〈0.01）, an elevated ratio of T-bet/GATA-3, and a decreased level of IL-10 mRNA in PBMC of N and T group compared to B group （P〈0.01）; and after treatment with Xiaoyin Recipe, IL-10 mRNA level increased （P〈0.01）, while TNF-α mRNA level and T-bet/GATA-3 ratio decreased in T group compared to N group （P〈0.01）. Conclusion： Xiaoyin Recipe for psoriasis could induce a Thl/Th2 balance drift toward Th2 in PBMC of EAT rats and thus improve the conditions.展开更多
OBJECTIVE:To observe the effect of herb-partitioned moxibustion(HPM)on the miRNA expression profile of thyroid tissue in experimental autoimmune thyroiditis(EAT)rats.METHODS:Rats were randomly divided into normal cont...OBJECTIVE:To observe the effect of herb-partitioned moxibustion(HPM)on the miRNA expression profile of thyroid tissue in experimental autoimmune thyroiditis(EAT)rats.METHODS:Rats were randomly divided into normal control(NC)group,EAT model(EAT)group,HPM group and western medicine(Med)group.EAT model rats were prepared by a combined immunization with complete and incomplete Freund’s adjuvant emulsified with porcine thyroglobulin and iodine.Rats in the HPM group were treated with HPM,while rats in the Med group were treated with levothyrocine(1μg/2 m L)by gavage.HE staining was used to observe the pathological morphological changes of thyroid tissue,ELISAs was uaed to detect the serum concentrations of TGAb,TPOAb,FT3,FT4,TSH.We then performed high-throughput mi RNA sequencing to analyse the mi RNA expression profiles in the thyroid tissues,followed by a bioinformatics analysis.RT-q PCR was used to verify the identified differentially expressed mi RNAs.RESULTS:HPM improved the thyroid tissue morphology and reduced serum TPOAb,TGAb,TSH concentration in EAT rats(P<0.05),but with no obvious effect on FT3 and FT4 concentration.While the TSH,FT3 and FT4 concentration was significantly changed in the Med group(P<0.01 or P<0.05)compared with that of EAT group.Sequencing results showed that a total of 17 mi RNAs were upregulated,and 4 were downregulated in the EAT rats,in which the expression levels of mi R-346 and mi R-331-5 p were reversed by HPM.The target genes of the mi RNAs that regulated by HPM wereassociated with a variety of immune factors and immune signals.RT-q PCR verification showed that the expression of mi RNA-346 and mi RNA-331-5 p was consistent with the sequencing results.CONCLUSIONS:HPM could regulate the the expression of mi RNA-346 and mi RNA-331-5 p,then act on their target genes to immune and inflammation-related pathways,which may be one of the mechanisms of HPM on EAT rats.展开更多
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...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。展开更多
基金supported by National Natural Science Foundation of China(No.81973855)。
文摘Objective:To investigate the effect of Fuzheng Xiaoying Decoction on experimental autoimmune thyroiditis(EAT)model rats and explore its possible mechanism of immune regulation.Methods:Among the 40 female SD rats,10 were randomly selected as the blank group,and the rest were immunized with pig thyroglobulin and Freund's adjuvant combined with high iodine feeding to make the EAT model.After the model was established,they were randomly divided into three groups:the model group,the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group,which were continuously gavaged for 2 months.The anti thyroid autoantibodies,thyroid function,IL-38 and Th17/Treg distribution were measured.The pathological changes of thyroid tissue were observed by HE staining.Results:Compared with the blank group,the levels of TPOAb and TGAb in the model group were significantly higher(P<0.05).Lymphocyte infiltration was seen in the thyroid tissue of the model group,and also the thyroid follicles were partially destroyed,the shape was irregular,and the colloid distribution was uneven,indicating that the modeling was successful.Compared with the model group,the levels of TPOAb and TGAb in the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group decreased significantly(P<0.05).And the Fuzheng Xiaoying Decoction group had more significant improvement in thyroid follicle destruction and lymphocyte infiltration than the selenium yeast tablet group(P<0.05).Compared with the blank group,the levels of T3,T4,FT3 and FT4 in the model group increased,the level of IL-38 decreased,the proportion of Th17 increased,and the ratio of Th17/Treg increased(P<0.05).Compared with the model group,T4 level decreased,IL-38 level increased in the selenium yeast tablet group and the Fuzheng Xiaoying Decoction group(P<0.05).But there was no statistically significant difference in the reduction of Th17 proportion and Th17/Treg ratio between the above two groups and the model group(P>0.05).There was no significant difference in Treg ratio among the groups.Conclusion:Fuzheng Xiaoying Decoction can significantly reduce the level of anti thyroid autoantibodies in EAT rats by regulating the immunity of EAT rats,and improve the follicular destruction and lymphocyte infiltration of thyroid tissue in rats.
文摘Objective： To explore the regulatory mechanism of Xiaoyin Recipe （消银方） on the T helper 1/ T helper 2 （Th1/Th2） immune balance. Methods： Thirty-six experimental animals were divided into three groups, 12 rats in each group： blank control group （B group）, negative control group （N group）, and Xiaoyin Recipe treatment group （T group）. The latter two groups received immunization of experimental autoimmune thyroiditis （EAT）, and T group were treated with Xiaoyin Recipe for a month. Then, the expression of Th1-Th2-related genes in peripheral blood mononuclear cells （PBMCs） were screened with Oligo GEArray Rat Thl-Th2-Th3 Microarray. The expressions of tumor necrosis factor- α （TNF- α）, interleukin-10 （IL-10）, T-box expressed in T-cells （T-bet）, and GATA-binding protein-3 （GATA-3） were detected by real-time polymerase chain reaction （RT-PCR）. Results： Gene array screening showed that compared to N group, in T group after Xiaoyin Recipe treatment, 3 genes were upregulated in EAT rats, including interleukin-27 receptor alpha （IL-27roL）, glomulin （GImn）, and GATA-3, while 38 genes were downregulated, such as CD28, IL-18, signal transducer, and activator of transcription 1 （STAT1）, T-bet, TNF receptor superfamily member 4 （TNFRSF4）, TNF ligand superfamily member 5 （TNFSF5）, and TNF receptor superfamily member 5 （TNFRSF5）. While RT-PCR showed that there was an increased level of TNF-α mRNA （P〈0.01）, an elevated ratio of T-bet/GATA-3, and a decreased level of IL-10 mRNA in PBMC of N and T group compared to B group （P〈0.01）; and after treatment with Xiaoyin Recipe, IL-10 mRNA level increased （P〈0.01）, while TNF-α mRNA level and T-bet/GATA-3 ratio decreased in T group compared to N group （P〈0.01）. Conclusion： Xiaoyin Recipe for psoriasis could induce a Thl/Th2 balance drift toward Th2 in PBMC of EAT rats and thus improve the conditions.
基金Supported by the National Natural Science Foundation of China(81704176,82074551)Three-year Action Plan Project of Shanghai Traditional Chinese Medicine Development(ZY(2018-2020)-CCCX-2004-01)+1 种基金the National Program on Key Basic Research Project(973 program)(2009CB522900,2015CB554501)Shanghai Municipal Health Commission Project(20194Y0013)。
文摘OBJECTIVE:To observe the effect of herb-partitioned moxibustion(HPM)on the miRNA expression profile of thyroid tissue in experimental autoimmune thyroiditis(EAT)rats.METHODS:Rats were randomly divided into normal control(NC)group,EAT model(EAT)group,HPM group and western medicine(Med)group.EAT model rats were prepared by a combined immunization with complete and incomplete Freund’s adjuvant emulsified with porcine thyroglobulin and iodine.Rats in the HPM group were treated with HPM,while rats in the Med group were treated with levothyrocine(1μg/2 m L)by gavage.HE staining was used to observe the pathological morphological changes of thyroid tissue,ELISAs was uaed to detect the serum concentrations of TGAb,TPOAb,FT3,FT4,TSH.We then performed high-throughput mi RNA sequencing to analyse the mi RNA expression profiles in the thyroid tissues,followed by a bioinformatics analysis.RT-q PCR was used to verify the identified differentially expressed mi RNAs.RESULTS:HPM improved the thyroid tissue morphology and reduced serum TPOAb,TGAb,TSH concentration in EAT rats(P<0.05),but with no obvious effect on FT3 and FT4 concentration.While the TSH,FT3 and FT4 concentration was significantly changed in the Med group(P<0.01 or P<0.05)compared with that of EAT group.Sequencing results showed that a total of 17 mi RNAs were upregulated,and 4 were downregulated in the EAT rats,in which the expression levels of mi R-346 and mi R-331-5 p were reversed by HPM.The target genes of the mi RNAs that regulated by HPM wereassociated with a variety of immune factors and immune signals.RT-q PCR verification showed that the expression of mi RNA-346 and mi RNA-331-5 p was consistent with the sequencing results.CONCLUSIONS:HPM could regulate the the expression of mi RNA-346 and mi RNA-331-5 p,then act on their target genes to immune and inflammation-related pathways,which may be one of the mechanisms of HPM on EAT rats.
文摘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。
