连花清瘟胶囊对脂多糖致急性肺损伤小鼠炎症因子和连接蛋白表达的影响

Effect of Lianhuaqingwen capsules on inflammatory cytokines and junction protein expression in mice with acute lung injury induced by lipopolysaccharides

目的研究连花清瘟胶囊(LHQW)对脂多糖(LPS)致急性肺损伤小鼠肺组织连接蛋白表达的影响。方法雄性KM小鼠随机分为6组,正常对照组、模型组、模型+地塞米松5 mg·kg-1组、模型+LHQW 2,4和8 g·kg-1组,每组20只。地塞米松和LHQW均ig给药,每天1次,共7 d。末次给药24 h后,除正常对照组外其余5组气管内滴注LPS溶液,制备小鼠急性肺损伤模型。造模24 h后,处死小鼠。光镜下观察肺组织病理形态变化,透射电镜下观察肺泡上皮超微结构,流式细胞术检测外周血T细胞中肿瘤坏死因子α(TNF-α)阳性表达细胞百分率,免疫组化法检测肺组织间隙连接蛋白43(Cx43)、闭锁蛋白和闭锁小带蛋白(ZO-1)的表达。结果光镜下观察发现,模型组小鼠肺内出现大量炎症细胞浸润,肺泡壁增厚;模型+地塞米松组、模型+LHQW 2,4和8 g·kg-1组较模型组炎症细胞浸润减少,肺泡壁增厚减轻。电镜下可见,模型组肺泡上皮细胞出现损伤,模型+地塞米松组、模型+LHQW 2,4和8 g·kg-1组较模型组均不同程度减轻。正常对照组、模型组、模型+地塞米松组、模型+LHQW 2,4和8 g·kg-1组外周血T细胞中TNF-α阳性表达细胞百分率分别为(3.6±0.9)%,(6.4±0.8)%,(2.8±0.7)%,(4.7±1.6)%,(4.0±1.5)%和(3.6±1.2)%,模型组明显高于正常对照组(P<0.01),其余各组均低于模型组(P<0.05,P<0.01)。模型组肺组织中Cx43、闭锁蛋白和ZO-1表达均低于正常对照组(P<0.01),模型+地塞米松、模型+LHQW 4和8 g·kg-1组3种蛋白表达均高于模型组(P<0.05)。结论 LHQW可能通过抑制炎症细胞浸润,改善肺泡上皮细胞和肺血管内皮细胞连接蛋白的表达,缓解LPS导致的急性肺损伤。

OBJECTIVE To explore the effect of Lianhuaqingwen capsules （ LHQW ） on junction＆amp;nbsp;protein expression in mouse lung tissue of lipopolysaccharide （LPS）-induced acute lung injury （ ALl）. METHODS 120 male mice were randomly divided into six groups： normal control, model, model＋dexa-methasone 5 mg.kg-1 , model ＋LHQW 2, 4 and 8 g.kg-1 groups. Dexamethasone and LHQW were administered orally, once daily, for 7 d. 24 h after the last administration, LPS solution was instilled into the tracheas of mice except the normal control group to prepare the mouse model of ALl. 24 h after the establishment of the ALl model, the mice were sacrificed and the pathological changes in the mouse lung tissue were observed by optical microscopy and ultrastructure of alveolar epithelium was observed by transmission electron microscopy. The cell percentage of positive expression of tumor necrosis factor-α（TNF-α） in the peripheral blood T lymphocytes was detected by flow cytometry. The expressions of con-nexin 43 （ Cx43）, occludin and zonula occludens protein-1 （ ZO-1） in lung tissues were detected by immunohistochemistry. RESULTS Under the light microscope, the mouse lung of model group showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Compared with model group, inflammatory cell infiltration was reduced in model＋dexamethasone, model＋LHQW 2,4 and 8 mg.kg-1 groups. Under the electron microscope, the mouse alveolar epithelial cells of model group showed injury. Compared with model group, the damage was reduced in model＋dexamethasone, and model＋LHQW 2, 4 and 8 mg.kg-1 groups. The cell percentage of TNF-α positive expression in peripheral blood T lympho-cytes in normal control, model, model＋dexamethasone, model＋LHQW 2,4 and 8 mg.kg-1 groups was （3.6±0.9）%, （6.4±0.8）%, （2.8±0.7）%, （4.7±1.6）%, （4.0±1.5）% and （3.6±1.2）%, respectively. The percentage in model group was obviously higher than that in normal control group（ P〈0.01）, but was lower in the four drug treatment groups than in model group（P〈0.05, P〈0.01）. The expression of Cx43, occludin and ZO-1 in lung tissue of model group was lower than that of normal control group（P〈0.01）, but higher in model＋dexamethasone, model ＋ LHQW 4 and 8 mg.kg-1 groups than in model group（P〈0.05）. CONCLUSION LHQW may alleviate ALl induced by LPS and play a protective role by inhibiting inflammatory cell infiltration and improving protein connection expression in alveolar epithelial cells and pulmonary vascular endothelial cells.