CAJ | 학술논문

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。
목적：연구련화청온효낭(LHQW)대지다당(LPS)치급성폐손상소서폐조직련접단백표체적영향。방법웅성 KM 소서수궤분위6조,정상대조조、모형조、모형+지새미송5 mg??kg-1조、모형+LHQW 2,4화8 g??kg-1조,매조20지。지새미송화 LHQW 균 ig 급약,매천1차,공7 d。말차급약24 h 후,제정상대조조외기여5조기관내적주 LPS 용액,제비소서급성폐손상모형。조모24 h 후,처사소서。광경하관찰폐조직병리형태변화,투사전경하관찰폐포상피초미결구,류식세포술검측외주혈 T 세포중종류배사인자α(TNF-α)양성표체세포백분솔,면역조화법검측폐조직간극련접단백43(Cx43)、폐쇄단백화폐쇄소대단백(ZO-1)적표체。결과광경하관찰발현,모형조소서폐내출현대량염증세포침윤,폐포벽증후；모형+지새미송조、모형+LHQW 2,4화8 g??kg-1조교모형조염증세포침윤감소,폐포벽증후감경。전경하가견,모형조폐포상피세포출현손상,모형+지새미송조、모형+LHQW 2,4화8 g??kg-1조교모형조균불동정도감경。정상대조조、모형조、모형+지새미송조、모형+ LHQW 2,4화8 g??kg-1조외주혈 T 세포중 TNF-α양성표체세포백분솔분별위(3.6±0.9)%,(6.4±0.8)%,(2.8±0.7)%,(4.7±1.6)%,(4.0±1.5)%화(3.6±1.2)%,모형조명현고우정상대조조(P<0.01),기여각조균저우모형조(P<0.05, P<0.01)。모형조폐조직중 Cx43、폐쇄단백화 ZO-1표체균저우정상대조조(P<0.01),모형+지새미송、모형+LHQW 4화8 g??kg-1조3충단백표체균고우모형조(P<0.05)。결론 LHQW 가능통과억제염증세포침윤,개선폐포상피세포화폐혈관내피세포련접단백적표체,완해 LPS 도치적급성폐손상。
OBJECTIVE To explore the effect of Lianhuaqingwen capsules ( LHQW ) on junction 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.