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嗜铬粒蛋白A衍生多肽CHR抑制LPS诱导的永生化脑微血管内皮细胞高通透性
Chromogranin A derived peptide CGA47-66 suppresses hyperpermeability of immortalized brain microvascular endothelial cells induced by lipopolysaccharide
- 第三军医大学学报 2018年40卷第15期 页码：1364-1369
- 作者机构：
400016 重庆, 重庆医科大学附属第一医院重症医学科
- 作者简介：
[ 张丹, E-mail:doctor_zhangdan@126.com ZHANG Dan, E-mail:doctor_zhangdan@126.com
- 基金信息：国家自然科学基金面上项目(81071531);国家自然科学基金面上项目(81372102);重庆市自然科学基金面上项目(CSTC2009BB5066)
DOI：10.16016/j.1000-5404.201801036
中图分类号： R322.13;R329.27;R341收稿：2018-01-04，
修回：2018-02-02，
纸质出版：2018-08-15
- 稿件说明：
移动端阅览
刘思佚, 曾燕, 刘疏柯, 等. 嗜铬粒蛋白A衍生多肽CHR抑制LPS诱导的永生化脑微血管内皮细胞高通透性[J]. 第三军医大学学报, 2018,40(15):1364-1369.
Siyi LIU, Yan ZENG, Shuke LIU, et al. Chromogranin A derived peptide CGA47-66 suppresses hyperpermeability of immortalized brain microvascular endothelial cells induced by lipopolysaccharide[J]. Journal of Third Military Medical University, 2018, 40(15): 1364-1369.
刘思佚, 曾燕, 刘疏柯, 等. 嗜铬粒蛋白A衍生多肽CHR抑制LPS诱导的永生化脑微血管内皮细胞高通透性[J]. 第三军医大学学报, 2018,40(15):1364-1369. DOI： 10.16016/j.1000-5404.201801036.
Siyi LIU, Yan ZENG, Shuke LIU, et al. Chromogranin A derived peptide CGA47-66 suppresses hyperpermeability of immortalized brain microvascular endothelial cells induced by lipopolysaccharide[J]. Journal of Third Military Medical University, 2018, 40(15): 1364-1369. DOI： 10.16016/j.1000-5404.201801036.
摘要
目的
2
探讨嗜铬粒蛋白A衍生多肽CHR（chromofungin）对LPS所诱导的永生化人脑微血管内皮细胞高通透性的影响。
方法
2
分别用LPS、CHR、CGA
4-16
作用于人脑微血管内皮细胞（human brain microvascular endothelial cells，HBMEC），用CCK8法检测不同浓度LPS刺激HBMEC细胞活力变化；EVOM法及Transwell小室法检测内皮细胞相对通透性；Western blot及RT-PCR法检测紧密连接蛋白（ZO-1和Claudin-5）的表达情况。
结果
2
与空白组相比，随着LPS浓度增加，HBMEC细胞的活性受到显著抑制（
P
<
0.05）。随着LPS作用时间延长，其细胞内紧密连接蛋白ZO-1、Claudin-5的表达逐渐下调，6 h后蛋白表达量降到最低（
P
<
0.05），12 h和24 h后其蛋白表达与6 h相比无明显差异。LPS作用下HBMEC单层细胞电阻值明显降低（
t
=-18.214，
P
<
0.001），细胞通透性显著升高（
t
=6.083，
P
=0.004），ZO-1、Claudin-5的蛋白和mRNA表达明显下调（
P
<
0.05），而CHR可显著改善上述变化。
结论
2
嗜铬粒蛋白A衍生多肽CHR能通过上调紧密连接相关蛋白的表达从而改善LPS所致的脑微血管内皮细胞高通透性。
Abstract
Objective
2
To determine the effect of chromogranin A derived peptide CGA47-66 (chromofungin
CHR) on lipopolysaccharide (LPS) induced high permeability of immortalized brain microvascular endothelial cells.
Methods
2
Immortalized human brain microvascular endothelial cells (HBMEC) were exposed to LPS
CHR and CGA4-16 respectively. Cell counting kit-8 (CCK8) assay was used to detect the cell viability after inducement of different concentrations of LPS. The relative permeability of HBMEC cells was measured by a Transwell chamber system and epithelial voltmeter (EVOM). The expression levels of the tight junction protein ZO-1 and Claudin-5 were detected by Western blotting and quantitative real-time polymerase chain reaction (RT-PCR).
Results
2
Compared with the blank control cells
the viability of HBMEC cells was significantly inhibited with the increase concentrations of LPS (
P
<
0.05). With the elapse of time
LPS treatment decreased the expression levels of ZO-1 and Claudin-5 gradually
and the levels reached the lowest after 6 h (
P
<
0.05)
which having no significant difference with the levels at 12 or 24 h. LPS treatment also resulted in significantly decreased transendothelial electrical resistance (
t
=-18.214
P
<
0.001)
increased cell permeability (
t
=6.083
P
=0.004)
and down-regulation of ZO-1 and Claudin-5 at mRNA and protein levels (
P
<
0.05). However
CHR significantly attenuated the above changes.
Conclusion
2
CHR could attenuate the hyperperneability of human brain microvascular endothelial cells induced by LPS through up-regulating tight junction proteins.
关键词
Keywords
references
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