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牦牛源CXCL14激活巨噬细胞的促炎反应并抑制HepG2细胞的增殖和迁移。

Yak-Derived CXCL14 Activates the Pro-Inflammatory Response of Macrophages and Inhibits the Proliferation and Migration of HepG2.

作者信息

Li Biao, Li Juan, Wang Li, Wei Yong, Luo Xiaolin, Guan Jiuqiang, Zhang Xiangfei

机构信息

Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China.

Key Laboratory of Animal Science of State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China.

出版信息

Animals (Basel). 2023 Sep 27;13(19):3036. doi: 10.3390/ani13193036.

Abstract

(C-X-C motif chemokine ligand 14) is an important chemokine involved in infection and immunity and plays an important role in a variety of immune-related diseases. The 446 bp cDNA sequence of the gene in yaks was obtained. Additionally, the prokaryotic expression vector of the CXCL14 protein with a molecular weight of 27 kDa was successfully constructed and expressed. The proliferation activities and migration abilities of spleen macrophages were significantly inhibited after treatment with the CXCL14 protein at different concentrations (1, 10 and 20 μg/mL) ( < 0.05). Furthermore, the expressions of pro-inflammatory cytokines interleukin 1 beta (), interleukin 6 (), interleukin 8 () and interferon-α () were significantly increased ( < 0.05), but the expression of anti-inflammatory factor interleukin 10 () was significantly decreased ( < 0.05). The contents of inflammatory factors in the supernatant of cells were detected using ELISA, and it was also found that the contents of TNF-α, IL6 and cytochrome c oxidase subunit II (COX2) were significantly increased under different CXCL14 protein concentrations ( < 0.05). Finally, the exogenous addition of CXCL14 inhibited the activity, clonal formation and migration of hepatoma cells (HepG2). Additionally, after HepG2 cells were treated with 20 μg/mL CXCL14 protein for 12 h, 24 h and 36 h, the expression levels of BCL2 homologous antagonist/killer () and the BCL2-associated X apoptosis regulator () were increased to varying degrees, while the expression levels of hypoxia-inducible factor 1 subunit alpha (, the mechanistic target of rapamycin kinase () and cyclin-dependent kinase 1 () genes decreased compared to the control group. In conclusion, the CXCL14 protein can inhibit the proliferation and migration of HepG2 cells by inducing the expression of macrophage pro-inflammatory factors and activating apoptosis-related genes to exert innate immunity. These results are helpful to further study the function of the CXCL14 protein and provide research data for the innate immune mechanism of yaks under harsh plateau environments.

摘要

(C-X-C基序趋化因子配体14)是一种参与感染和免疫的重要趋化因子,在多种免疫相关疾病中发挥重要作用。获得了牦牛该基因446 bp的cDNA序列。此外,成功构建并表达了分子量为27 kDa的CXCL14蛋白的原核表达载体。用不同浓度(1、10和20 μg/mL)的CXCL14蛋白处理后,脾巨噬细胞的增殖活性和迁移能力受到显著抑制(P<0.05)。此外,促炎细胞因子白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素8(IL-8)和干扰素-α(IFN-α)的表达显著增加(P<0.05),但抗炎因子白细胞介素10(IL-10)的表达显著降低(P<0.05)。采用ELISA法检测细胞上清液中炎症因子含量,发现在不同CXCL14蛋白浓度下肿瘤坏死因子-α(TNF-α)、IL-6和细胞色素c氧化酶亚基II(COX2)含量均显著增加(P<0.05)。最后,外源性添加CXCL14抑制了肝癌细胞(HepG2)的活性、克隆形成和迁移。此外,用20 μg/mL CXCL14蛋白处理HepG2细胞12 h、24 h和36 h后,BCL2同源拮抗剂/杀手(BAK)和BCL2相关X凋亡调节因子(BAX)的表达水平有不同程度升高,而缺氧诱导因子1亚基α(HIF-1α)、雷帕霉素激酶机制靶点(mTOR)和细胞周期蛋白依赖性激酶1(CDK1)基因的表达水平与对照组相比降低。综上所述,CXCL14蛋白可通过诱导巨噬细胞促炎因子表达和激活凋亡相关基因来抑制HepG2细胞增殖和迁移,从而发挥固有免疫作用。这些结果有助于进一步研究CXCL14蛋白的功能,为牦牛在恶劣高原环境下的固有免疫机制提供研究数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5bb3/10571970/63ff6bc234d5/animals-13-03036-g001.jpg

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