参附注射液对脂多糖所致肺泡细胞损伤的保护作用

doi:10.3877/cma.j.issn.1674-6880.2021.05.003

中华危重症医学杂志(电子版) ›› 2021, Vol. 14 ›› Issue (05) : 362 -367. doi: 10.3877/cma.j.issn.1674-6880.2021.05.003

论著

参附注射液对脂多糖所致肺泡细胞损伤的保护作用

乔莉¹, 赵超¹, 孙昊¹, 陈洁², 王军³, 张劲松¹^,^†(

)

^1. 210029　南京，南京医科大学第一附属医院急诊科
^2. 210011　南京，南京医科大学第二附属医院重症医学科
^3. 211166　南京，南京医科大学公共卫生学院

收稿日期:2021-01-13 出版日期:2021-10-31
通信作者: 张劲松
基金资助:
国家自然科学基金项目(81772057、81571875); 江苏省医学创新团队项目(CXTDA2017007); 北京协和医学基金"睿E（睿意）急诊医学研究专项基金"(R2013002)

Protective effects of Shenfu injection on alveolar cell injury induced by lipopolysaccharide

Li Qiao¹, Chao Zhao¹, Hao Sun¹, Jie Chen², Jun Wang³, Jinsong Zhang¹^,^†()

^1. Department of Emergency Medicine, the First Affiliated Hospital with Nanjing Medical University, Nanjing 210029, China
^2. Intensive Care Unit, the Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China
^3. School of Public Health, Nanjing Medical University, Nanjing 211166, China

Received:2021-01-13 Published:2021-10-31
Corresponding author: Jinsong Zhang

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目的

探讨参附注射液（SFI）对脂多糖（LPS）所致肺泡细胞损伤的保护作用及其对通透性调控相关蛋白血管内皮钙黏蛋白（VE-cadherin）、ZO-1、Claudin-5和细胞间连接黏附分子2（JAM-2）的影响。

方法

将MLE-12细胞分为对照组、LPS组、SFI组和SFI + LPS组。LPS组和SFI + LPS组均采用2.0 μg/mL的LPS处理MLE-12细胞4 h，以制作急性脓毒症细胞模型。SFI组和SFI + LPS组均采用100 μL/mL的SFI预处理2 h，对照组仅培养基培养6 h。采用细胞计数试剂盒（CCK-8）法测定各组细胞活力，Western-blotting检测各组细胞VE-cadherin、ZO-1、Claudin-5及JAM-2蛋白表达水平。

结果

4组细胞间VE-cadherin、ZO-1、Claudin-5、JAM-2蛋白表达水平及细胞活力比较，差异均有统计学意义（F = 98.000、73.060、59.104、83.007、81.206，P = 0.043、0.007、0.003、0.023、0.001）。进一步两两比较发现，与对照组比较，LPS组VE-cadherin、ZO-1、Claudin-5、JAM-2蛋白表达水平和细胞活力均显著降低，SFI组上述蛋白表达水平均显著升高；与LPS组比较，SFI和SFI + LPS组VE-cadherin、ZO-1、Claudin-5、JAM-2蛋白表达水平和细胞活力均显著升高，且SFI组更高（P均< 0.05）。

结论

SFI能减轻LPS所致的小鼠肺泡上皮细胞中膜通透性调控相关蛋白表达的下调作用，对LPS所致的肺泡通透性增加效应有一定的治疗作用。

关键词: 参附注射液, 脂多糖, 小鼠肺泡上皮细胞株

Objective

To study the effect of Shenfu injection (SFI) on alveolar cell injury caused by lipopolysaccharide (LPS), and to study the mechanism of SFI on vascular endothelial-cadherin (VE-cadherin), ZO-1, Claudin-5 and junctional adhesion molecule-2 (JAM-2) which are related to permeability regulation.

Methods

MLE-12 cells were divided into a control group, a LPS group and a SFI + LPS group. To make an acute sepsis cell model, MLE-12 cells were induced with 2.0 μg/mL LPS for 4 h in the LPS group and SFI + LPS group. The SFI group and SFI + LPS group were pretreated with 100 μL/mL SFI for 2 h, and the control group was only cultured with medium for 6 h. Cell viability of each group was measured by the cell counting kit-8, and the protein expression levels of VE-cadherin, ZO-1, Claudin-5 and JAM-2 were analyzed by Western-blotting.

Results

There were statistically significant differences in the VE-cadherin, ZO-1, Claudin-5 and JAM-2 protein expression levels and cell viability among the four groups (F = 98.000, 73.060, 59.104, 83.007, 81.206; P = 0.043, 0.007, 0.003, 0.023, 0.001). Compared with the control group, the VE-cadherin, ZO-1, Claudin-5 and JAM-2 protein expression levels and cell viability in the LPS group significantly decreased, while the protein expression levels in the SFI group significantly increased. Compared with the LPS group, the VE-cadherin, ZO-1, Claudin-5 and JAM-2 protein expression levels and cell viability in SFI and SFI + LPS groups significantly increased, which were highest in the SFI group (all P < 0.05).

Conclusion

SFI could attenuate the down-regulation of permeability related proteins, and have a certain therapeutic effect on the increased alveolar permeability caused by LPS.

Key words: Shenfu Injection, Lipopolysaccharide, Mice alveolar epithelial cell line

图1 CCK-8检测不同浓度LPS和SFI对MLE-12细胞的活力影响（n = 3）

图2 不同时间点LPS对VE-cadherin、ZO-1蛋白表达的调节作用（n = 3）

图3 各组MLE-12细胞间VE-cadherin、ZO-1、Claudin-5及JAM-2蛋白表达水平的比较（n = 3）

图4 各组MLE-12细胞活力的比较（n = 3）

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Protective effects of Shenfu injection on alveolar cell injury induced by lipopolysaccharide

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Protective effects of Shenfu injection on alveolar cell injury induced by lipopolysaccharide