CAJ | 학술논문

目的观察维生素E联煤系厝姿啥孕∈蠹毙苑嗡鹕搜字⒓凹∏虻鞍浊崃醇っ父(MLCK)表达的影响.方法将40只8周龄Balb/c雌性小鼠按随机数字表法分为4组:生理盐水对照组(1.5 ml/kg)、细菌脂多糖组(1 mg/kg)、维生素E联合地塞米松治疗组(联合治疗组)、维生素E治疗组(维生素E组).联合治疗组给予腹腔注射维生素E 50 mg/kg和地塞米松1 mg/kg,1 h后再给予脂多糖1 mg/kg滴鼻,维生素E组仅给予维生素E脂质体50 mg/kg,观察各组小鼠肺组织病理改变.采用免疫组织化学SABC法检测肺组织MLCK免疫反应细胞,用逆转录-PCR反应检测肺组织中MLCK mRNA的表达,Western blot法检测肺组织中MLCK蛋白的表达.各组间均数比较采用单因素方差分析,均数两两比较采用SNK检验.结果脂多糖组小鼠的肺部炎症反应显著,以中性粒细胞浸润为主,伴明显的肺泡充血、水肿.两治疗组肺组织炎症、充血明显改善;对照组、脂多糖组、联合治疗组、维生素E组BALF中细胞总数分别为(1.1±0.4)、(5.6±2.1)、(4.0±1.0)、(4.2±1.3)×109/L,脂多糖组与其他3组比较,差异有统计学意义(F=14.53,均P<0.05).脂多糖组MLCK免疫反应细胞较对照组明显增加,主要在上皮和内皮,联合治疗组和维生素E组MLCK免疫反应细胞明显减少;脂多糖组与两治疗组肺组织MLCK mRNA吸光度值比较,差异无统计学意义(F=2.76,均P>0.05);脂多糖组与两治疗组肺组织MLCK蛋白吸光度值比较,差异有统计学意义(F=12.06,均P<0.01).结论维生素E联合地塞米松可抑制脂多糖诱导的小鼠急性肺损伤肺部炎症及肺组织MLCK蛋白的表达,其作用可能部分通过抑制MLCK活性而达到稳定血管屏障、改善肺水肿和炎症.
목적 관찰유생소E련매계조자사잉∈두폐원옹호수자⒓요∏맹안탁래순っ부(MLCK)표체적영향.방법 장40지8주령Balb/c자성소서안수궤수자표법분위4조:생리염수대조조(1.5 ml/kg)、세균지다당조(1 mg/kg)、유생소E연합지새미송치료조(연합치료조)、유생소E치료조(유생소E조).연합치료조급여복강주사유생소E 50 mg/kg화지새미송1 mg/kg,1 h후재급여지다당1 mg/kg적비,유생소E조부급여유생소E지질체50 mg/kg,관찰각조소서폐조직병리개변.채용면역조직화학SABC법검측폐조직MLCK면역반응세포,용역전록-PCR반응검측폐조직중MLCK mRNA적표체,Western blot법검측폐조직중MLCK단백적표체.각조간균수비교채용단인소방차분석,균수량량비교채용SNK검험.결과 지다당조소서적폐부염증반응현저,이중성립세포침윤위주,반명현적폐포충혈、수종.량치료조폐조직염증、충혈명현개선;대조조、지다당조、연합치료조、유생소E조BALF중세포총수분별위(1.1±0.4)、(5.6±2.1)、(4.0±1.0)、(4.2±1.3)×109/L,지다당조여기타3조비교,차이유통계학의의(F=14.53,균P<0.05).지다당조MLCK면역반응세포교대조조명현증가,주요재상피화내피,연합치료조화유생소E조MLCK면역반응세포명현감소;지다당조여량치료조폐조직MLCK mRNA흡광도치비교,차이무통계학의의(F=2.76,균P>0.05);지다당조여량치료조폐조직MLCK단백흡광도치비교,차이유통계학의의(F=12.06,균P<0.01).결론 유생소E연합지새미송가억제지다당유도적소서급성폐손상폐부염증급폐조직MLCK단백적표체,기작용가능부분통과억제MLCK활성이체도은정혈관병장、개선폐수종화염증.
Objective To observe the effect of vitamin E (VitE) combined with dexamethasone (DXM) on inflammation of acute lung injury and expression of myosin light chain kinase. Methods Forty female Balb/c mice were randomly divided into 4 groups, a saline control group(1.5 ml/kg), a LPS group (1 mg/kg), a VitE and DXM group(VitE 50 mg/kg, DXM 1 mg/kg), and a VitE group(50 mg/kg). Lung tissue histopathological changes were observed. Immunohistochemistry assays (SABC) were used to determine the myosin light chain kinase(MLCK) immunoreactive cells in the lung tissues, and the MLCK mRNA and the MLCK protein was assayed by RT-PCR and by Western blot, respectively. Means were compared with analysis of variance and Student-Newman-Keuls were used to compare 2 means. Results Histological examination showed that extensive lung inflammation were seen in the LPS group, which manifested by accumulation of significant numbers of neutrophils, accompanied by marked pulmonary edema and hemorrhage. The inflammation and hemorrhage in the 2 treatment groups were significantly improved. Immunoreactive cells of MLCK numbers in BALF in the control group, the LPS group, the VitE and DXM group, and the VitE group was (1.1±0.4), (5.6±2.1), (4.0±1.0), (4.2±1.3)×10<'9>/L respectively. Compared with other groups, the difference of LPS group was significant(F =14.53, all P<0.05). Immunoreactive cells of MLCK located airway epithelial and endothelia in the LPS group were more than which in the control group, decreased immunoreactive cells of MLCK in two treatment groups. Compared with LPS group, the difference of MLCK mRNA expression (A) of lung tissue in two treatment DOI: 10.3760/cma.j.issn.1001-0939.2009.01.013groups was no significant(F=2.76,all P>0.05). Compared with LPS group, the difference of A values of MLCK protein of lung tissue in two treatment groups was statistical significance (F = 12.06, all P<0.01). Conclusions Vitamin E combined with low dose of DXM could effectively inhibit inflammation and expression of MLCK protein in acute lung injury induced by LPS lung. It is suggested that, inhibition of MLCK activation leads to stabilize vascular barrier function and attenuation of pulmonary edema and inflammation, which also suggests a possible role of MLCK in the pathogenesis of acute lung injury.