The rCC16 Protein Protects Against LPS-Induced Cell Apoptosis and Inflammatory Responses in Human Lung Pneumocytes
The rCC16 Protein Protects Against LPS-Induced Cell Apoptosis and Inflammatory Responses in Human Lung Pneumocytes
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rCC16蛋白对lps诱导的人肺细胞凋亡和炎症反应的保护作用
DOI：
10.3389/fphar.2020.01060
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发表时间：
2020-07-14
影响因子：
5.6
通讯作者：
Zeng, Xiaobin
中科院分区：
文献类型：
--
作者：
Lin, Jinle;Li, Jiemei;Zeng, Xiaobin
目的本课题组前期临床研究表明，肺内CC16水平降低对ARDS的发生、发展有重要影响。本研究的目的是评估rCC16对LPS诱导的A549细胞炎症的治疗作用并确定其机制。方法采用脂多糖（LPS）刺激诱导细胞凋亡和炎症反应。采用MTT法检测rCC16的细胞毒作用。使用酶联免疫吸附试验测定细胞因子水平。通过分析相关信号通路，探讨rCC16的分子机制。结果LPS处理后A549细胞存活率明显降低，凋亡蛋白Bax、巴克和Cleaved Caspase-3表达增加，炎性细胞因子分泌增加，TLR4、p-NF/kappa B、MAPK蛋白表达增加。LPS处理后A549细胞Bcl-2、p-AKT、p-mTOR、p-ERK 1/2、NF/kappa B、p-AMPK和p-p38表达明显降低。我们的实验结果还证实rCC 16抑制LPS诱导的细胞凋亡，通过激活PI3 K/AKT/mTOR/ERK 1/2通路促进A549细胞增殖，通过TLR4/NF-κ B/AMPK信号通路的去磷酸化和失活抑制某些炎症因子的释放，特别是HMG B1。结论CC16作为一种重要的细胞因子，具有潜在的抗炎、抗炎作用，在ARDS的临床治疗中可能发挥重要作用。
Objective Our previous clinical study showed that low lung levels of CC16 strongly influence the occurrence and development of ARDS. The aim of the present study was to evaluate the therapeutic effect of rCC16 on LPS-induced inflammation in A549 cells and to determine its mechanism. Methods Cell apoptosis and inflammation was induced by LPS stimulation. The cytotoxic effect of rCC16 was evaluated using the MTT assay. Cytokine levels were determined using enzyme-linked immunosorbent assays. The molecular mechanism of rCC16 was investigated by analyzing relevant signaling pathways. Results The LPS treatment of A549 cells significantly decreased cell viability, increased the levels of the apoptotic proteins Bax, Bak and Cleaved Caspase-3, the secretion of inflammatory cytokines, and the expression levels of TLR4, p-NF/kappa B, MAPK proteins. While the levels of Bcl-2, p-AKT, p-mTOR, p-ERK1/2, NF/kappa B, p-AMPK, and p-p38 were significantly decreased in LPS-treated A549 cells. Our experimental results also confirmed that rCC16 inhibited LPS-induced apoptosis, promoted A549 cell proliferation by activating the PI3K/AKT/mTOR/ERK1/2 pathway, and inhibited the release of certain inflammatory factors, especially HMGB1, through dephosphorylation and inactivation of the TLR4/NF-kappa B/AMPK signaling pathways. Conclusion These results highlight the potential utility of CC16 as an important cytokine for the prevention or treatment of inflammation and show that CC16 may play an important role in the future clinical treatment of ARDS.