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河北省衡水市哈励逊国际和平医院内分泌科,衡水,053000
纸质出版日期:2021,
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侯丽萍. 维生素D3对实验性自身免疫性甲状腺炎Th1/Th2细胞平衡和MCP-1/CCR2信号的影响[J]. 重庆医科大学学报, 2021,46(8):921-926.
Hou Liping. Effects of vitamin D3on helper T cell 1/helper T cell 2 cell balance and monocyte chemoattractant protein-1/cell surface chemokine receptor 2 signal in experimental autoimmune thyroiditis[J]. Journal of Chongqing Medical University, 2021,46(8):921-926.
侯丽萍. 维生素D3对实验性自身免疫性甲状腺炎Th1/Th2细胞平衡和MCP-1/CCR2信号的影响[J]. 重庆医科大学学报, 2021,46(8):921-926. DOI: 10.13406/j.cnki.cyxb.002524.
Hou Liping. Effects of vitamin D3on helper T cell 1/helper T cell 2 cell balance and monocyte chemoattractant protein-1/cell surface chemokine receptor 2 signal in experimental autoimmune thyroiditis[J]. Journal of Chongqing Medical University, 2021,46(8):921-926. DOI: 10.13406/j.cnki.cyxb.002524.
目的: 探讨维生素D3对实验性自身免疫性甲状腺炎大鼠炎症、辅助性T细胞1/辅助性T细胞2(helper T cell 1/helper T cell 2,Th1/Th2)及单核细胞趋化蛋白1/细胞表面趋化因子受体2(monocyte chemoattractant protein-1/cell surface chemokine receptor 2,MCP-1/CCR2)信号的影响。方法: 将Lewis大鼠按随机数字表法分为对照组(n=10)、模型组(n=15)和治疗组(n=15),采用免疫猪甲状腺球蛋白(porcine thyroglobulin,pTG)诱导建立实验性自身免疫性甲状腺炎(experimental autoimmune thyroiditis,EAT)大鼠模型,治疗组按照每只5 mg/kg的剂量腹腔注射1,25-二羟维生素D3[1,25-(OH)2D3],隔天1次,共用4周,其余2组均给予等体积生理盐水。采用苏木素-伊红(hematoxylin-eosin stain,HE)染色观察大鼠甲状腺组织形态学变化;酶联免疫吸附法(enzyme-linked immunosorbentassay,ELISA)检测血清中促甲状腺激素(thyroid stimulating hormone,TSH)、自身抗体抗甲状腺球蛋白抗体(anti-thyroglobulin antibody,TGAb)、甲状腺过氧化物酶抗体(anti-thyroid peroxidase antibody,TPOAb)及细胞因子干扰素-γ(interferon,IFN-γ)、白细胞介素-12(interleukin-12,IL-12)、白细胞介素-4(interleukin-4,IL-4)、白细胞介素-10(interleukin-10,IL-10),逐步线性回归分析TSH与自身抗体、细胞因子的关系;免疫组织化学检测甲状腺组织中的NF-κB p65;Western blot检测甲状腺组织中MCP-1/CCR2信号轴。结果: 模型组甲状腺滤泡结构破坏、炎性细胞浸润,治疗组甲状腺病理变化明显改善并趋向于对照组。治疗组TSH、TGAb、TPOAb水平与模型组相比均明显降低(t=4.000、2.603、5.279,P=0.000、0.015、0.000),但仍明显高于对照组(t=2.400、2.216、7.392,P=0.025、0.037、0.000)。与模型组相比,治疗组IL-4、IL-10水平明显升高(t=3.522、2.432,P=0.001、0.022),IFN-γ、IL-12水平及IFN-γ/IL-4、IL-12/IL-10比值明显降低(t=2.940、4.700、5.416、8.178,P=0.007、0.000、0.000、0.000),均趋向于对照组水平。逐步回归分析结果显示,TSH与TGAb、TPOAb、IFN-γ、IL-12均呈正相关,与IL-4和IL-10呈负相关(r=0.872、0.868、0.731、0.706、-0.557、-0.236,均P<0.001),其中TGAb、TPOAb和IL-12对TSH的影响最明显。与模型组比较,治疗组大鼠甲状腺组织中的NF-κB p65、MCP-1及CCR2水平均下降,差异具有统计学意义(t=2.432、2.267、5.143,P=0.000、0.031、0.000)。结论: 维生素D3可能是通过抑制NF-κB通路以下调Th1型细胞因子、上调Th2型细胞因子进而降低Th1/Th2比值,还可能是通过抑制MCP-1/CCR2信号轴,最终减轻EAT大鼠甲状腺炎,改善甲状腺功能,抑制甲状腺抗体生成,保护甲状腺。
Objective: To investigate the effect of vitamin D3on inflammation
helper T cell 1/helper T cell 2 (Th1/Th2) and monocyte chemoattractant protein-1/cell surface chemokine receptor 2 (MCP-1/CCR2) signal in experimental autoimmune thyroiditis (EAT) rats. Methods: Lewis rats were randomly divided into the control group (n=10)
the model group (n=15) and the treatment group (n=15). The rat model of EAT was established by immunizing porcine thyroglobulin (pTG). The treatment group received intraperitoneal injection of 1
25- (OH) 2vitamin D3at the dose of 5 mg/kg
once every other day for consecutive four weeks
and the other two groups were given normal saline of equal volume. The morphological changes of thyroid tissue in rats were observed by hematoxylineosin (HE) staining. Serum thyroid stimulating hormone (TSH)
anti-thyroglobulin antibody (TGAb)
anti-thyroid peroxidase antibody (TPOAb) and cytokines interferon-γ (IFN-γ)
interleukin-12 (IL-12)
interleukin-14 (IL-4) and interleukin-10 (IL-10) were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between TSH and autoantibodies and cytokines was analyzed by stepwise linear regression. NF-κB p65 in thyroid tissue was detected by immunohistochemistry. MCP-1/CCR2 signal axis in thyroid tissue was analyzed by Western blot. Results: In the model group
thyroid follicles were damaged and inflammatory cells were infiltrated
while the pathological changes of thyroid in the treatment group were significantly improved and tended to the control group. The levels of TSH
TGAb and TPOAb in the treatment group were significantly lower than those in the model group (t=4.000
2.603
5.279; P=0.000
0.015
0.000)
but were still significantly higher than those in the control group (t=2.400
2.216
7.392; P=0.025
0.037
0.000). Compared with the model group
the levels of IL-4 and IL-10 were significantly increased in the treatment group (t=3.522
2.432; P=0.001
0.022)
and the levels of IFN-γ
IL-12 and IFN-γ/IL-4 and IL-12/IL-10 were significantly decreased in the treatment group (t=2.940
4.700
5.416
8.178; P=0.007
0.000
0.000
0.000)
which tended to the levels of the control group. Stepwise regression analysis showed that TSH was positively correlated with TGAb
TPOAb
IFN-γ and IL-12
and negatively correlated with IL-4 and IL-10 (r=0.872
0.868
0.731
0.706
-0.557
-0.236
all P<0.001)
among which TGAb
TPOAb and IL-12 had the most significant effect on TSH. Compared with the model group
the levels of NF-κB p65
MCP-1 and CCR2 in the thyroid tissues of rats in the treatment group were decreased
with statistically significant differences (t=2.432
2.267
5.143; P=0.000
0.031
0.000). Conclusion: Vitamin D3may inhibit NF-κB pathway to down-regulate Th1 cytokines and up-regulate Th2 cytokines
thereby reducing Th1/Th2 ratio. Or
it reduces thyroiditis in EAT rats
improves thyroid function
inhibits thyroid antibody production and protects thyroid by inhibiting MCP-1/CCR2 signaling axis.
维生素D3实验性自身免疫性甲状腺炎辅助性T细胞1/辅助性T细胞2单核细胞趋化蛋白1/细胞表面趋化因子受体2
vitamin D3experimental autoimmune thyroiditishelper T cell 1/helper T cell 2monocyte chemoattractant protein-1/cell surface chemokine receptor 2
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