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布雷他汀改善高位胆总管结扎术大鼠肺炎性损伤的研究
Blebbistatin ameliorates pneumonic injury after high common bile duct ligation in rats
- 陆军军医大学学报 2022年44卷第8期 页码：757-764
- 作者机构：
400038 重庆，陆军军医大学(第三军医大学)第一附属医院麻醉科
- 作者简介：
[鲁开智，E-mail:lukaizhi2010@163.comLU Kaizhi, E-mail: lukaizhi2010@163.com
- 基金信息：国家自然科学基金面上项目(81671961)
DOI：10.16016/j.2097-0927.202202013
中图分类号： R563.05;R575;R977.6收稿：2022-02-08，
修回：2022-03-03，
纸质出版：2022-04-30
- 稿件说明：
移动端阅览
赖静, 陈林, 李才溢, 等. 布雷他汀改善高位胆总管结扎术大鼠肺炎性损伤的研究[J]. 陆军军医大学学报, 2022,44(8):757-764.
Jing LAI, Lin CHEN, Caiyi LI, et al. Blebbistatin ameliorates pneumonic injury after high common bile duct ligation in rats[J]. Journal of Army Medical University, 2022, 44(8): 757-764.
赖静, 陈林, 李才溢, 等. 布雷他汀改善高位胆总管结扎术大鼠肺炎性损伤的研究[J]. 陆军军医大学学报, 2022,44(8):757-764. DOI： 10.16016/j.2097-0927.202202013.
Jing LAI, Lin CHEN, Caiyi LI, et al. Blebbistatin ameliorates pneumonic injury after high common bile duct ligation in rats[J]. Journal of Army Medical University, 2022, 44(8): 757-764. DOI： 10.16016/j.2097-0927.202202013.
摘要
目的
2
探讨非肌肉肌球蛋白Ⅱ(non-muscle myosin Ⅱ
NMⅡ)抑制剂布雷他汀对慢性胆总管结扎术后肺组织炎性损伤的保护作用。
方法
2
选用体质量200~220 g的SPF级雄性SD大鼠55只，按照随机数字表法分为对照组(Sham组，
n
=11)、高位胆总管结扎术(common bile duct ligation
CBDL)模型组(CBDL组，
n
=22)和高位胆总管结扎术后布雷他汀治疗组[CBDLB组，
n
=22，腹腔注射布雷他汀5 mg/kg(PEG-300∶生理盐水=1∶1为溶剂)
]
。通过HE染色观察肺组织的改变；Western blot检测肺组织MPO、IL-6、TNF-α、SOD3表达情况；免疫组化检测炎症相关的MPO、IL-6、IL-1β、TNF-α、LY6B、CD163因子和血管内皮相关因子CD31的表达水平。
结果
2
胆总管结扎术后3周肺部损伤表现为大量的炎性细胞浸润为主，伴有局部肺组织肺泡腔和肺间质出血；免疫组化结果显示CBDL组相较于Sham组MPO、LY6B和CD163阳性细胞数量均显著增多(
P
<
0.01)
同时炎症因子IL-1β、TNF-α的表达量也显著升高(
P
<
0.01)；Western blot检测结果也同样证实CBDL组相较于Sham组肺组织炎性因子MPO、IL-6和超氧化物歧化酶SOD3分泌增多(
P
<
0.05)；胆总管结扎术后5周肺组织损伤主要表现为肺微血管扩张，CBDL组相较于Sham组肺微血管扩张明显(
P
<
0.01)。胆总管结扎的大鼠经过布雷他汀治疗1周后，第3周免疫组化结果显示CBDLB组较CBDL组MPO、LY6B和CD163阳性细胞数量均显著减少(
P
<
0.01)，同时炎症因子IL-1β、IL-6和TNF-α的表达量也显著降低(
P
<
0.01)，Western blot检测结果显示CBDLB组较CBDL组炎性因子IL-6、MPO和TNF-α的分泌减少(
P
<
0.05);第5周CBDLB组较CBDL组肺微血管扩张程度明显减轻(
P
<
0.01)。
结论
2
布雷他汀能有效抑制胆总管结扎后肺组织炎性细胞的浸润，减轻肺炎症损伤，缓解胆总管结扎后期肺微血管扩张。
Abstract
Objective
2
To investigate the protective effect of blebbistatin
a non-muscle myosin Ⅱ (NMⅡ) inhibitor
on inflammatory injury of lung tissue after chronic common bile duct ligation (CBDL) in rats.
Methods
2
A total of 55 SPF male SD rats (200~220 g) were randomly divided into 3 groups
namely control group (Sham
n
=11)
high CBDL group (model group
n
=22) and blebbistatin treatment after high CBDL group (treatment group
n
=22). The treatment group was intraperitoneally injected with blebbistatin at a dose of 5 mg/kg once a day for a week
while the model group was only given the solvent (PEG-300∶saline=1∶1). The changes of lung tissue were observed with HE staining. Western blotting was performed to detect the expression of myeloperoxidase (MPO)
IL-6
TNF-α
and superoxide dismutase (SOD3) in the lung tissues. The expression levels of MPO
IL-6
IL-1β
TNF-α
LY6B
CD163
and vascular endothelium related factor CD31 were also determined by immunohistochemical assay.
Results
2
The lung injury at the 3rd week after CBDL presented a large number of inflammatory cells infiltration
accompanied by local alveolar space and pulmonary interstitial hemorrhage. Immunohistochemical results demonstrated that the numbers of MPO positive cells
LY6B positive cells and CD163 positive cells were all increased significantly in the model group as compared with the Sham group (
P
<
0.01)
with enhanced expression of IL-1β and TNF-α (
P
<
0.01). Western blotting results confirmed the up-regulated levels of MPO
IL-6 and SOD3 in the model group than the sham group (
P
<
0.05). In comparison with the model group
the treatment group showed decreased numbers o
f MPO positive cells
LY6B positive cells
and CD163 positive cells (
P
<
0.01)
and declined levels of inflammatory factors IL-1β
IL-6
and TNF-α (
P
<
0.01). Western blotting indicated as well that the expression of MPO
IL-6 and TNF-α in the treatment group was reduced greatly (
P
<
0.05). Moreover
at the 5th week after surgery
the lung injury mainly manifested as pulmonary microvascular dilation
which was more severe in the model group than the Sham group (
P
<
0.01)
while the severity of pulmonary microvascular dilatation was remarkably milder after treatment (
P
<
0.01).
Conclusion
2
Blebbistatin can effectively inhibit the infiltration of inflammatory cells in lung tissue
reduce pneumonia injury and alleviate pulmonary microvascular dilatation after CBDL.
关键词
Keywords
references
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