Smad3 gene C-terminal phosphorylation site mutation aggravates CCl4-induced inflammation in mice
Smad3 gene C-terminal phosphorylation site mutation aggravates CCl4-induced inflammation in mice
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Smad3基因C端磷酸化位点突变加重CCl4诱导的小鼠炎症
DOI：
10.1111/jcmm.15385
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发表时间：
2020-05-14
影响因子：
5.3
通讯作者：
Yang, Yan
中科院分区：
文献类型：
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作者：
Ding, Hanyan;Fang, Meng;Yang, Yan
随着肝病的进展，C末端磷酸化Smad3(PSmad3C)的表达下调。因此，我们假设pSmad3C的表达可能与肝脏疾病负相关。为了开发新的治疗策略，需要一个合适的动物模型，使研究人员能够研究Smad3结构域特异性磷酸化对肝脏疾病进展的影响。本研究旨在构建一种新的带有Smad3 C末端磷酸化位点突变的小鼠模型，并探讨该突变在CCl4诱导的炎症反应中的作用。采用TetraOne(TM)基因定点敲入技术和胚胎干细胞显微注射技术获得Smad3 C末端磷酸化位点突变小鼠。得到的小鼠通过基因分型进行鉴定，并在存在或不存在CCl4的情况下探索其对炎症的影响。没有纯合子的小鼠出生，这表明这种突变是胚胎致命的。在没有试剂刺激的情况下，野生型(WT)和杂合子(HT)小鼠的肝脏表型和生长没有显著差异。在CCl4诱导的急性和慢性肝损伤后，HT小鼠的肝脏病理、血清转氨酶(ALT/AST)表达和炎症因子(IL-6/TNF-α)水平的改变比WT小鼠更严重。此外，HT组小鼠肝组织中pSmad3C蛋白水平较低。这些结果提示Smad3 C末端的磷酸化可能在肝损伤的早期阶段起到保护作用。总之，我们已经建立了一个新的动物模型，它将成为未来研究Smad3结构域特异性磷酸化对肝脏疾病进展影响的新工具。
The expression of C-terminal phosphorylated Smad3 (pSmad3C) is down-regulated with the progression of liver disease. Thus, we hypothesized that pSmad3C expression may be negatively related to liver disease. To develop novel therapeutic strategies, a suitable animal model is required that will allow researchers to study the effect of Smad3 domain-specific phosphorylation on liver disease progression. The current study aimed to construct a new mouse model with the Smad3 C-terminal phosphorylation site mutation and to explore the effects of this mutation on CCl4-induced inflammation. Smad3 C-terminal phosphorylation site mutant mice were generated using TetraOne (TM) gene fixed-point knock-in technology and embryonic stem cell microinjection. Resulting mice were identified by genotyping, and the effects on inflammation were explored in the presence or absence of CCl4. No homozygous mice were born, indicating that the mutation is embryonic lethal. There was no significant difference in liver phenotype and growth between the wild-type (WT) and heterozygous (HT) mice in the absence of reagent stimulation. After CCl4-induced acute and chronic liver damage, liver pathology, serum transaminase (ALT/AST) expression and levels of inflammatory factors (IL-6/TNF-alpha) were more severely altered in HT mice than in WT mice. Furthermore, pSmad3C protein levels were lower in liver tissue from HT mice. These results suggest that Smad3 C-terminal phosphorylation may have a protective effect during the early stages of liver injury. In summary, we have generated a new animal model that will be a novel tool for future research on the effects of Smad3 domain-specific phosphorylation on liver disease progression.