摘要
目的 探讨非肌肉肌球蛋白Ⅱ(non-muscle myosinⅡ, 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 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 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 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。
Objective 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 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, LY6 B, CD163, and vascular endothelium related factor CD31 were also determined by immunohistochemical assay. Results The lung injury at the 3 rd 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, LY6 B 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 of MPO positive cells, LY6 B 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 5 th week after surgery, the lung injury mainly manifested as pulmonary microvascular d
作者
赖静
陈林
李才溢
王丹丹
鲁开智
LAI Jing;CHEN Lin;LI Caiyi;WANG Dandan;LU Kaizhi(Department of Anesthesiology,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400038,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2022年第8期757-764,共8页
Journal of Army Medical University
基金
国家自然科学基金面上项目(81671961)。
关键词
布雷他汀
炎症
慢性胆总管结扎
肝肺综合征
blebbistatin
inflammation
chronic common bile duct ligation
hepatopulmonary syndrome
