黄芩苷对NCI-H292细胞凋亡和LPS所致小鼠急性肺损伤的影响  

Effects of baicalin on NCI-H292 cell apoptosis and acute lung injury induced by lipopolysaccharide

作　　者：李慧峰 单明辉[2] 程淑琴 高建新[4] 

机构地区：[1]河北省北方学院动物科技学院,河北张家口075000 [2]河北省廊坊市大厂畜牧水产局,河北廊坊065300 [3]河北省旅游职业学院,河北承德067000 [4]河北省邯郸市广平县综合职业技术教育中心,河北邯郸057650

出　　处：《中国兽医学报》2014年第5期811-815,824,共6页Chinese Journal of Veterinary Science

基　　金：河北省高等学校科学技术研究基金资助项目(Z2011117);张家口市科技攻关计划资助项目(12110037C-2)

摘　　要：为探讨黄芩苷对肺上皮细胞(NCI-H292细胞)凋亡和脂多糖(lipopolysaccharide,LPS)所致小鼠急性肺损伤的影响。本试验采用Hoechst染色法和流式细胞法检测黄芩苷(60和120μg/L)对H2O2诱导NCI-H292细胞凋亡率的影响;腹腔注射LPS建立小鼠急性肺损伤模型,观察黄芩苷对LPS诱导的小鼠急性肺损伤的影响。将ICR小鼠随机分为4组,分别为空白对照组、LPS组、黄芩苷150mg/(kg·d)组和300mg/(kg·d)组,予以蒸馏水或黄芩苷灌胃,1次/d,连续3d,于试验第3d灌胃后1h,除空白对照组其他各组腹腔注射LPS(20mg/(kg·d)),造模12h后观察肺组织病理学变化,测定肺组织匀浆中髓过氧化物酶(MPO)的活力,用Western blotting检测肺组织胞浆型磷脂酶A2(cPLA2)及磷酸化cPLA2的含量。结果显示,Hoechst染色可见黄芩苷60和120μg/L预处理对H2O2诱导NCI-H292细胞凋亡均有保护作用;流式细胞法测定显示,与无黄芩苷刺激的H2O2组相比,黄芩苷作用后呈剂量依赖抑制NCI-H292细胞凋亡。小鼠腹腔注射LPS后12h,肺组织明显充血、水肿,并有大量炎性细胞浸润,肺组织MPO、cPLA2水平显著升高。与LPS组相比,黄芩苷的两个剂量均能显著降低MPO和cPLA2的水平,明显减轻内毒素所致的肺损伤。结果表明,黄芩苷对H2O2诱导的肺上皮细胞NCI-H292凋亡具有保护作用,能通过降低肺组织中MPO含量和cPLA2活性,减轻小鼠内毒素性肺损伤。To explore the effects of baicalin on apoptosis of lung epithelial cell and acute lung inju- ry induced by lipopolysaccharide（LPS）. Apoptotic rates of lung epithelial cells induced by hydro- gen peroxide were measured by Hoechst staining and flow cytometry. LPS was used to establish acute lung injury model of mice by intraperitoneal injection to observe the effects of baicalin on a- cute lung injury. ICR mice were randomly divided into 4 groups, blank control,LPS,LPS＋baica- lin（150 mg/（kg ; d）） and LPS＋ baicalin（300 mg/（kg ; d））. Mice were administered intragastri- cally with distilled water or baicalin once a day for 3 days. One hour after intragastrical treatment on the third day, LPS （20 mg/kg） was injected intraperitoneally except blank control group. Twelve hours after establishing the animal model, the histological changes of lungs were ob- served, myeloperoxidase （MPO） activity in lung homogenate was measured, and cytosolic phos- pholipase A2 （cPLA2） and phosphorylated cytosolic phospholipase A2 （phosphor-cPLA2） in lung tissues were detected by Western blotting. The results showed baicalin （60 and 120μg/L）de-creased the apoptosis of NCI-H292 cells induced by H202 ,which was measured by Hoechst stai- ning. Compared with the model group,baicalin concentration dependently suppressed the apopto- sis of NCI-H292 cells measured by flow cytometry. Twelve hours after LPS challenge, Histologi- cal studied showed that there were congestion, edema and the infiltrate of inflammatory cells in lung tissues in LPS group, and MPO activity and the levels of cPLA2 in lung homogenate were highly increased. Instead lung injury was significantly alleviated in two dose baicalin ; LPS groups. Baicalin has significant anti-apoptotic effects and could relieve LPS -induced acute lung injury by diminishing the contents of MPO and cPLA2 both in vitro and in vivo.

关 键 词：黄芩苷 细胞凋亡 流式细胞法 脂多糖 急性肺损伤 髓过氧化物酶 磷脂酶A 

分 类 号：S853.7[农业科学—临床兽医学]