益气活血方对机械通气肺损伤大鼠肺上皮Ⅱ型细胞的保护机制研究  

Effect of Yiqi Huoxue Decoction on Type Ⅱ Lung Epithelial Cells in Rats with Ventilator Induced Lung Injury

作　　者：李瑾 宋文龙 Lin Jin;Song Wenlong(Jiande Integrated Traditional Chinese and Western Medicine Hospital,Zhejiang,Jiande 311600,China)

机构地区：[1]浙江省建德市中西医结合医院,浙江建德311600

出　　处：《中国中医急症》2023年第7期1159-1162,1170,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：杭州市卫生科技计划项目(0020190760);建德市科技发展计划基金项目(2018JYW12)。

摘　　要：目的 观察益气活血方对机械通气肺损伤(VLLI)大鼠肺泡上皮细胞中Ghrelin、JNK、ERK、p38、核转录因子-κB(NF-κB)、IKβ-α及炎症因子表达的变化等,以探讨其对VLLI大鼠肺上皮Ⅱ型细胞的保护机制。方法 以SD大鼠肺上皮Ⅱ型细胞为研究对象,采用机械牵拉方式模拟机械通气肺损伤,予益气活血方含药血清进行干预。ELISA检测各组大鼠肺上皮Ⅱ型细胞Ghrelin、TNF-α、白细胞介素-6(IL-6)表达,Western blotting检测p-JNK、p-ERK、p-p38蛋白表达水平,RT-PCR法检测JNK、ERK、p38、NF-κB、IKβ-α、Ghrelin的mRNA相对表达量。结果 经机械牵拉损伤后,大鼠肺上皮Ⅱ型细胞中IL-6、TNF-α较前明显上升,且随刺激时间延长表达水平升高;电镜下大鼠肺上皮Ⅱ型细胞形态学发生显著的改变;与对照组比较,机械牵拉损伤大鼠肺上皮Ⅱ型细胞中p-JNK、p-ERK、p-p38蛋白和mRNA,以及NF-κB基因表达均有不同程度升高,Ghrelin、IKβ-α基因相对表达水平降低(P <0.05或P <0.01)。给药后,和模型组相比,中药组p-JNK、p-ERK、p-p38蛋白和mRNA以及NF-κB基因表达水平下降,Ghrelin、IKβ-α mRNA表达水平上调(P> 0.05)。结论 益气活血方对VLLI模型大鼠肺上皮Ⅱ型细胞有保护作用,其作用可能是通过上调Ghrelin的表达来阻断MAPK/NF-κB通路,从而缓解炎症反应,使炎症细胞因子表达减少。Objective:To observe the effect of Yiqi Huoxue Decoction on the expression of Chrelin,JNK,ERK,p38,NF-kB,IKβ-αand inflammatory factors in alveolar epithelial cells of rats with ventilator induced lung injury(VILI),and to elucidate its possible mechanism of protecting lung tissue.Methods:The lung epithelial type Ⅱ cells of SD rats were used as the research subjects.VILI model was simulated by mechanical stretch.The serum containing Yiqi Huoxue Decoction was intervened.Ghrelin,TNF-α,and IL-6 were detected by ELISA,the expres-sions of p-JNK,p-ERK and p-p38 protein were detected by Western boltting,and the relative mRNA expressions of JNK,ERK,p38,Ghrelin,NF-kB and IKβ-αwere detected by RT-PCR.Results:After mechanical stretch inju-ry,the expression of IL-6 and TNF-αin type I cells of rat's lung epithelium was significantly increased,and the expression level increased with the stimulation time.Under the electron microscope,the morphological changes of type Ⅱ cells were observed;compared with the control group,the p-JNK,p-ERK,p-p38 protein and mRNA,as well as NF-kB gene expression levels increased to varying degrees in type Ⅱ cells of lung epithelium in rats with VILI,while the levels of ghrelin and IKβ-αrelative gene expression reduced(P<0.05 or P<0.01).After adminis-tration,compared with the model group,the p the p-JNK,p-ERK,p-p38 protein and mRNA,as well as NF-kB gene expression levels in the TCM group decreased,while the expression of Ghrelin and IKβ-αexpression levels increased(P>0.05).Conclusion:Yiqi Huoxue Decoction has a protective effect on type Ⅱ lung epithelial cells in VLLI model rats,whose mechanism of action is related to increased of the expression of Ghrelin and blocking MAPK/NF-kB pathway,alleviateion inflammatory reaction,and reduction the expression of inflammatory cytokines.

关 键 词：机械通气肺损伤 益气活血方 肺上皮Ⅱ型细胞 保护作用 MAPK/NF-κB通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]