内质网应激对肝泡型包虫病部分炎症介质的影响

李姚; 王靖超; 袁加琪; 温浩; 周瀛; 樊海宁; 王志鑫

doi:10.3969/j.issn.1001-5256.2021.01.026

内质网应激对肝泡型包虫病部分炎症介质的影响

DOI: 10.3969/j.issn.1001-5256.2021.01.026

1.
青海大学附属医院肝胆胰外科，西宁 810000
2.
新疆医科大学第一附属医院消化血管外科中心肝胆包虫外科，乌鲁木齐 830000

基金项目:

青海省重大科技专项子课题 (2016-SF-A5);

2017年青海“高端创新人才千人计划” (培养拔尖人才项目)

利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突，特此声明。

作者贡献声明：李姚、王靖超、袁加琪、温浩、周瀛、樊海宁、王志鑫负责课题设计、资料分析; 李姚、王靖超负责收集数据与撰写论文; 李姚、王靖超、王志鑫参与修改论文; 温浩、周瀛、樊海宁、王志鑫责拟定写作思路，指导撰写论文并最后定稿。

详细信息

作者简介:
李姚(1992—)，男，主要从事肝胆胰外科、肝包虫病学研究

王靖超(1994—)，男，主要从事肝胆胰外科、肝包虫病学研究

通信作者:
王志鑫，wangzhixin126@163.com

二人对本文贡献相同，同为第一作者

中图分类号: R575;R535
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出版历程
- 收稿日期: 2020-05-20
- 录用日期: 2020-09-09
- 出版日期: 2021-01-20

Influence of endoplasmic reticulum stress on the secretion of some inflammatory mediators in hepatic alveolar echinococcosis

1.
Department of Hepatopancreatobiliary Surgery, Qinghai University Affiliated Hospital, Xining 810000, China
2.
Department of Hepatobiliary and Hydatid Surgery, Center of Digestive and Vascular Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, China

摘要

摘要: 目的探究内质网应激(ERS)在肝泡型包虫病(HAE)病情发展过程中对部分炎症介质的影响，初步探讨其临床意义。方法选取2018年6月—2019年9月因HAE在青海大学附属医院接受肝脏部分切除术的患者15例，切除HAE病灶边缘带作为AE组，同期肝脏正常组织15例作为对照组。通过蛋白质印迹法、实时荧光定量PCR检测AE组和对照组中的蛋白激酶R样激酶(PERK)、CCAAT/增强子结合蛋白同源蛋白(CHOP)、含半胱氨酸的天冬氨酸蛋白水解酶12(caspase-12)和葡萄糖调节蛋白78(GRP-78)表达水平以及IL-1β、IL-6和TNF的mRNA表达水平。两组间不满足正态性的计量资料比较采用Mann-Whitney U检验; 满足正态性的计量资料两组间分析采用t检验; 双变量相关性分析采用Pearson相关分析法。结果 AE组中PERK、CHOP、caspase-12、GRP-78的蛋白表达量(U值分别为4.165、3.461、2.577、3.344，P值均＜0.001)和mRNA表达水平(t值分别为34.003、4.461、53.573、55.224，P值均＜0.001)均明显高于对照组，差异均有统计学意义。AE组中IL-1β、IL-6和TNF的mRNA表达水平明显高于对照组(t值分别为6.090、12.578、53.573，P值均＜0.001)。PERK、CHOP、caspase-12、GRP-78的蛋白表达水平与IL-1β、IL-6和TNF的mRNA表达水平均呈正相关(r值均＞0.700，P值均＜0.05)。结论 HAE中ERS的激活与炎症介质的活化存在正相关性，ERS的过度激活能够改变部分炎症介质的分泌来加剧肝脏的损伤，具体机制有待进一步研究。
- 棘球蚴病, 肝 /
- 内质网应激 /
- 炎症 /
- 细胞因子类
Abstract: Objective To investigate the influence of endoplasmic reticulum stress (ERS) on some inflammatory mediators during the progression of hepatic alveolar echinococcosis (HAE) and its clinical significance. Methods A total of 15 patients with HAE who underwent partial liver resection in Qinghai University Affiliated Hospital from June 2018 to September 2019 were enrolled, and the marginal zone of HAE lesion was resected as AE group; 15 normal liver tissue samples collected during the same period of time were selected as control group. Western blot and qRT-PCR were used to measure the protein and mRNA expression of protein kinase R-like ER kinase (PERK), CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12, and glucose-regulated protein-78 (GRP-78), and q-PCR was used to measure the mRNA expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF). The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, and the t-test was used for comparison of normally distributed continuous data between two groups; a Pearson correlation analysis was performed to investigate the correlation between two variables. Results Compared with the control group, the AE group had significantly higher protein expression levels of PERK, CHOP, caspase-12, and GRP78 (U=4.165, 3.461, 2.577, and 3.344, all P < 0.001) and their mRNA expression levels (t= 34.003, 4.461, 53.573, and 55.224, all P < 0.001). The AE group had significantly higher mRNA expression levels of IL-1β, IL-6, and TNF than the control group (t=6.090, 12.578, and 53.573, all P < 0.001). The protein expression levels of PERK, CHOP, caspase-12, and GRP-78 were positively correlated with the mRNA expression levels of IL-1β, IL-6, and TNF (all r > 0.700, all P≤0.05). Conclusion Positive correlation is observed between the activation of ERS and inflammatory mediators in HAE, and excessive activation of ERS can change the secretion of several inflammatory mediators to exacerbate liver injury, while further studies are needed to clarify the specific mechanism.
- Echinococcosis, Hepatic /
- Endoplasmic Reticulum Stress /
- Inflammation /
- Cytokines

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表 1 PCR引物序列

基因	引物序列(5′-3′)	引物长度(bp)
IL-1β	R：CGATCACTGAACTGCACGCTC	168
	F：ACAAAGGACATGGAGAACACCACTT
IL-6	R：CAATGAGGAGACTTGCCTGGTG	203
	F：TGGCATTTGTGGTTGGGTCA

下载: 导出CSV

表 2 两组间ERS相关蛋白表达水平的比较

组别	例数	PERK	CHOP	caspase-12	GRP-78
AE组	15	27.63(25.32~29.32)	37.36(35.34~39.52)	31.41(29.34~34.63)	30.89(28.34~33.12)
对照组	15	19.89(18.34~21.28)	28.63(24.52~32.14)	20.36(17.62~24.83)	20.45(16.34~23.66)
U值		4.165	3.461	2.577	3.344
P值		＜0.001	＜0.001	＜0.001	＜0.001

下载: 导出CSV

表 3 两组间ERS相关指标mRNA表达水平的比较

组别	例数	PERK	CHOP	caspase-12	GRP-78
AE组	15	27.445±0.553	37.007±1.364	31.252±0.568	30.927±0.536
对照组	15	19.892±0.658	27.752±2.539	20.227±0.559	20.110±0.138
t值		34.003	4.461	53.573	55.224
P值		＜0.001	＜0.001	＜0.001	＜0.001

下载: 导出CSV

表 4 两组间相关炎症介质mRNA表达水平的比较

组别	例数	IL-1β	IL-6	TNF
AE组	15	3.265±1.215	33.461±5.991	26.368±2.472
对照组	15	1.294±0.311	12.548±2.361	10.563±0.618
t值		6.090	12.578	53.573
P值		＜0.001	＜0.001	＜0.001

下载: 导出CSV

表 5 ERS相关蛋白与炎症介质的相关性

指标	PERK		CHOP		caspase-12		GRP-78
指标	r值	P值	r值	P值	r值	P值	r值	P值
IL-1β	0.811	0.016	0.782	0.008	0.778	＜0.001	0.786	0.005
IL-6	0.922	0.020	0.879	0.032	0.934	0.012	0.917	＜0.001
TNF	0.784	＜0.001	0.943	0.011	0.935	0.006	0.885	0.031

下载: 导出CSV

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