AjTGFβ alleviates V. splendidus-induced inflammation through SMADs pathway in Apostichopus japonicus.
AjTGFβ alleviates V. splendidus-induced inflammation through SMADs pathway in Apostichopus japonicus.
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DOI：
10.1016/j.fsi.2023.108593
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发表时间：
2023-02
影响因子：
4.7
通讯作者：
Zhan Zhen;Ye Wenwen;Han Guanghui;L. Chenghua;Zhimeng Lv
中科院分区：
文献类型：
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作者：
Zhan Zhen;Ye Wenwen;Han Guanghui;L. Chenghua;Zhimeng Lv
抑制炎症反应是控制炎症发展的必要过程，是保护机体免受过度炎症损伤的重要步骤。转化生长因子β（transforming growth factor beta，TGF-β）作为一种多效性细胞因子，在脊椎动物体内起着抑制炎症的调节作用。为了研究TGF-β在无脊椎动物炎症反应中的调控作用，本研究采用cDNA末端快速扩增的方法克隆了仿刺参（Apostichopus）TGF-β基因，命名为AjTGF-β。而在刺激后96 h，体腔细胞中AjTGF-β mRNA的表达量下降了0.27倍，与炎症反应的趋势相反。AjTGF-β在所有组织中均有表达，体壁中表达最高。当使用小干扰RNA（siRNA-KD）将AjTGF-β敲低至0.45倍时，AjSMAD 2/3和AjSMAD 6与对照组相比分别下调至0.32倍和0.05倍。与对照组相比，重组AjTGF-β与灿烂弧菌共孵育后，损伤区无广泛炎症反应，损伤修复时间为72 h，AjSMAD 2/3表达上调1.35倍。在此条件下，AjSMAD 2/3沉默减轻rAjTGF-β诱导的损伤恢复。与Vs + BSA组相比，rAjTGF-β可轻微诱导I型胶原表达，从6.13 ng/mL上调至7.84 ng/mL，III型胶原表达从6.23 ng/mL上调至6.89 ng/mL。这一发现表明AjTGF-β通过AjSMADs调节胶原蛋白表达来负性调节炎症进展并加速损伤修复。
The inhibition of inflammatory response is an essential process to control the development of inflammation and is an important step to protect the organism from excessive inflammatory damage. As a pleiotropic cytokine, transforming growth factor beta (TGF-β) plays a regulatory role in inhibiting inflammation in vertebrates. To investigate the role of TGF-β in the regulation of inflammation in invertebrates, we cloned and characterized the TGF-β gene from Apostichopus japonicus via rapid amplification of cDNA ends, and the sample was designated as AjTGF-β. For Vibrio splendidus-challenged sea cucumbers, the expression of AjTGF-β mRNAs in coelomocytes decreased at 96 h (0.27-fold), which was contrary to the trend of inflammation. AjTGF-β was expressed in all tissues with the highest expression in the body wall. When AjTGF-β was knocked down by using small interfering RNA (siRNA-KD) to 0.45-fold, AjSMAD 2/3 and AjSMAD6 were downregulated to 0.32-and 0.05-fold compared with the control group, respectively. Furthermore, when the damaged sea cucumber was challenged by V. splendidus co-incubated with rAjTGF-β, the damage area had no extensive inflammation, and damaged repair appeared at 72 h compared with the Vs+ BSA group, in which the expression of AjSMAD 2/3 was upregulated by 1.35-fold. Under this condition, AjSMAD 2/3 silencing alleviated rAjTGF-β-induced damage recovery. Moreover, rAjTGF-β slightly induced the collagen I expression from 6.13 ng/mL to 7.84 ng/mL, and collagen III was upregulated from 6.23 ng/mL to 6.89 ng/mL compared with the Vs+ BSA group. This finding indicates that AjTGF-β negatively regulated the inflammatory progress and accelerated the repair of damage by AjSMADs to regulate the collagens expression.