跨膜蛋白TMEM88在LPS/D-GaIN诱导的急性肝损伤中的作用

何翠霞; 朱敏慧; 徐媛媛; 王雪枫; 丁佳翔; 周焕

doi:10.16766/j.cnki.issn.1674-4152.003503

跨膜蛋白TMEM88在LPS/D-GaIN诱导的急性肝损伤中的作用

doi: 10.16766/j.cnki.issn.1674-4152.003503

何翠霞^{1, 2},
朱敏慧^{1, 2},
徐媛媛^{1, 2},
王雪枫^{1, 2},
丁佳翔^{1, 3},
周焕^{1, 2, 3, ,}

1.
蚌埠医科大学第一附属医院国家药物临床试验机构，安徽蚌埠 233004
2.
蚌埠医科大学药学院，安徽蚌埠 233030
3.
蚌埠医科大学公共基础学院，安徽蚌埠 233030

基金项目:

蚌埠医科大学“512人才培育计划”优秀青年教师基金项目 by51201313

详细信息

通讯作者:
周焕，E-mail: zhouhuan@bbmc.edu.cn

中图分类号: R575 R-332
计量
- 文章访问数: 35
- HTML全文浏览量: 26
- PDF下载量: 3
- 被引次数: 0
出版历程
- 收稿日期: 2023-02-15
- 网络出版日期: 2024-07-20

Role of the transmembrane protein TMEM88 in acute liver injury induced by LPS/D-GaIN

HE Cuixia^{1, 2},
ZHU Minhui^{1, 2},
XU Yuanyuan^{1, 2},
WANG Xuefeng^{1, 2},
DING Jiaxiang^{1, 3},
ZHOU Huan^{1, 2, 3
, ,}

1.
Clinical Trials Center, the First Affiliated Hospital of Bengbu Medical University, Bengbu, Anhui 233004, China

摘要

摘要: 目的探讨脂多糖(LPS)联合D-半乳糖胺(D-GaIN)诱导急性肝损伤小鼠中TMEM88的表达变化，并研究TMEM88表达高低对LPS诱导的AML-12细胞炎症因子分泌的影响。方法体内，将20只小鼠按随机数字表法随机分为正常组与模型组，每组10只，模型组采用LPS/D-GaIN联合诱导，收集小鼠肝脏组织和血液。体外，AML-12细胞经LPS刺激及TMEM88转染，测定TMEM88及炎症因子的表达变化。结果模型组小鼠肝脏组织发生明显损伤，ALI模型造模成功率为80%。此外，模型组小鼠肝脏TMEM88表达水平均明显升高(P<0.05)。在AML-12细胞中，LPS刺激的最适条件为100 ng/mL，刺激24 h。pEGFP-C1-TMEM88和TMEM88-siRNA经验证成功转染进LPS刺激的AML-12细胞内(P<0.05)。此外，TMEM88过表达会导致炎症因子IL-1β、IL-6及TNF-α分泌上调(P<0.01)，而敲低TMEM88则会减少炎症因子的分泌(P<0.05)，以上实验结果表明TMEM88的表达水平可能影响炎症过程。结论 TMEM88的表达水平在LPS/D-GaIN诱导的急性肝损伤模型中具有重要作用。
- TMEM88 /
- LPS/D-GaIN /
- 急性肝损伤 /
- 炎症因子
Abstract: Objective This study aims to investigate the changes of TMEM88 expression in mice with acute liver injury induced by lipopolysaccharide (LPS) combined with D-galactosamine (D-GaIN), and investigate the effect of high and low TMEM88 expression on the secretion of inflammatory factors in AML-12 cells induced by LPS. Methods In vivo, 20 mice were randomly divided into normal and model groups with 10 mice in each group according to random number table method. The model group was induced by LPS/D-GaIN combination, and liver tissues and blood of mice were collected. In vitro, AML-12 cells were stimulated by LPS and transfected with TMEM88 to access the changes in the expression of TMEM88 and inflammatory factors. Results The model group exhibited significant liver tissue, with a successful ALI modelling rate of 80%. In addition, the hepatic TMEM88 expression level was significantly increased in the model group (P<0.05). In AML-12 cells, the optimal condition for LPS stimulation was determined to be 100 ng/mL for 24 h. pEGFP-C1-TMEM88 and TMEM88-siRNA were verified to be successfully transfected into LPS-stimulated AML-12 cells (P<0.05). Furthermore, overexpression of TMEM88 resulted in an upregulation of inflammatory factors IL-1β, IL-6, and TNF-α (P<0.01), while knockdown of TMEM88 reduced the secretion of inflammatory factors (P<0.05). These experimental results suggest that the expression level of TMEM88 may influence the inflammatory process. Conclusion The expression level of TMEM88 plays an important role in the LPS/D-GaIN-induced acute liver injury model.
- TMEM88 /
- LPS/D-GaIN /
- Acute liver injury /
- Inflammatory factor

HTML全文

图 1 ALI模型构建及TMEM88的表达

注：A为ALI及正常肝脏HE染色；B和C为RT-qPCR法及免疫印迹法检测ALI及正常肝脏中TMEM88的表达；与正常肝脏组织比较，^aP<0.05，^bP<0.01。

Figure 1. Construction of ALI models and TMEM88 expression

下载: 全尺寸图片幻灯片

图 2 LPS刺激对AML-12细胞中TMEM88表达的影响

注：A为CCK8实验检测细胞存活率，^aP<0.05；B和C为免疫印迹法测定LPS不同浓度及刺激时长下TMEM88水平，与正常组比较，^bP<0.05，^cP<0.05；D为100 ng/mL LPS诱导24 h后TMEM88水平，与正常组比较，^dP<0.05。

Figure 2. Impact of LPS stimulation on TMEM88 expression in AML-12 cells

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图 3 TMEM88敲低和过表达模型构建

注：A和B为RT-qPCR法及免疫印迹法敲低TMEM88，与TMEM88-NC组比较，^aP<0.01；C和D为RT-qPCR法及免疫印迹法过表达TMEM88，与pEGFP-C1组比较，^bP<0.01。

Figure 3. Construction of TMEM88 knockdown and overexpression models

下载: 全尺寸图片幻灯片

图 4 TMEM88过表达调控炎症因子的分泌

注：A为RT-qPCR法及免疫印迹法检测TMEM88过表达对炎症因子表达的影响，与pEGFP-C1比较，P<0.05；B为敲低TMEM88对炎症因子表达的影响，与TMEM88-NC比较，P<0.05。

Figure 4. Overexpression of TMEM88 regulates the secretion of inflammatory cytokines

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. Primer sequence

项目	序列
TMEM88	F: 5'-CCTGTGCAGTGCTAGTAACGG-3'
	R: 5'-GCCGAAGCCTAACATGATGACA-3'
GAPDH	F: 5'-AGGTCGGTGTGAACGGATTTG-3'
	R: 5'-GGGGTCGTTGATGGCAACA-3'
IL-1β	F: 5'-TTCAGGCAGGCAGTATCACTC-3'
	R: 5'-GAAGGTCCACGGGAAAGACAC-3'
TNF-α	F: 5'- CAGGCGGTGCCTATGTCTC-3'
	R: 5'-CGATCACCCCGAAGTTCAGTAG-3'
IL-6	F: 5'-CTGCAAGAGACTTCCATCCAG-3'
	R: 5'-AGTGGTATAGACAGGTCTGTTGG-3'

下载: 导出CSV

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