转化生长因子β1对原代培养猪甲状腺过氧化物酶活性及mRNA表达的影响

目的 观察转化生长因子(TGF)-β1对原代培养猪甲状腺过氧化物酶(TPO)及mRNA表达的影响,探讨其可能的作用机制.方法 体外培养猪甲状腺细胞,按照TGF-β1水平,实验分为0μg/L(对照组),2、5、10μg/L组,培养72 h后,用噻唑蓝(MTT)法测定TGF-β1对细胞存活率,用愈创木酚法测定TGF-β1对细胞TPO活性的影响,用反转录聚合酶链反应(RT-PCR)方法检测甲状腺细胞TPOmRNA表达.结果 ①与对照组(100%)比较,2、5、10μg/L组细胞的存活率(85.26%、75.14%、63.21%)均明显降低(P均<0.05).②与对照组(2.143±0.102)比较,2、5、10μg/L组的TPO活性[(1.915±0.047)、(1.423±0.073)、(1.018±0.101)]均明显下降(P均<0.05),呈显著的负相关(r=-0.979,P<0.01).③与对照组(1.419±0.148)比较,2、5、10μg/L组的TPO mRNA表达[(0.875±0.078)、(O.466±0.044)、0.273±0.007)]明显降低(P均<0.05).结论 TGF-β1对原代培养猪甲状腺细胞有抑制的作用,可能与TGF-β1抑制甲状腺细胞TPO活性及其mRNA表达有关.

Abstract：

Objective To observe the effect of transforming growth factor β1 (TGF-β1) on thyroid peroxidase activity and the expression of mRNA in primary porcine thyrocytes, and approach its possible mechanism. Methods In vitro cultured porcine thyrocytes were divided into 0 μg/L(control group), 2, 5, and 10 μg TGF-β1/L groups.After cultured for 72 h, the cell survival rate was measured with methyl thiazolyl tetrazolium (MTT) assay and the thyroid peroxidase (TPO) activity was measured by guaiacol method. The expression of TPO mRNA was determined cell survival rates of 2, 5, 10 μg/L groups(85.26%, 75.14% and 63.21%) were significantly lower(all P < 0.05 ).significantly decreased as compared with the control group [(2.143 ± 0.102), all P < 0.05)], and the TPO of TGF-β1, the lower the expression of TPO mRNA [(0.875 ± 0.078), (0.466 ± 0.044), (0.273 ± 0.007)] compared with the control group[(1.419 ± 0.148), all P < 0.05)]. Conclusions The inhibiting effect of TGF-β1 on primary cultured porcine thyrocytes may be related to its mechanism of inhibiting TPO activity and reduced mRNA expression.