补肺汤对HMGB1诱导的特发性肺纤维化相关细胞TLR2/NF-κB信号通路的影响  

Effects of Bufei Decoction on TLR2/NF-κB Signal Pathway in HMGB1-Induced Idiopathic Pulmonary Fibrosis Cells

作　　者：文秀华[1] 杨红军[1] WEN Xiu-hua;YANG Hong-jun(Department of Geriatrics,Sichuan No.2 Hospital of Traditional Chinese Medicine,Chengdu,610031)

机构地区：[1]四川省第二中医医院老年病科,成都610031

出　　处：《中国中西医结合杂志》2021年第10期1228-1234,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：四川省公益性科研院所基本科研项目(No.2019-4-529)。

摘　　要：目的观察补肺汤对高迁移率族蛋白B1(HMGB1)诱导的人非小细胞肺癌细胞(A549)和人肺成纤维细胞(HFL1)形态损伤、炎症反应和Toll样受体2/髓样分化蛋白88/核因子-κB(TLR2/MyD88/NF-κB)信号通路的影响。方法使用冷冻技术制备补肺汤冻干粉,大鼠补肺汤灌胃4周后取含药血清。将A549细胞和HFL1细胞随机分为空白组、HMGB1组(HMGB1诱导)、补肺汤冻干粉预培养加HMGB1组(冻干粉组,冻干粉浓度分别为0.10%、0.15%、0.20%、0.25%、0.30%)、补肺汤含药血清预培养加HMGB1组(含药血清组,含药血清浓度分别为5%、10%、15%、20%、25%)、阳性药物组(甲泼尼松龙)及空白血清组,按照分组处理细胞。电镜下观察各组细胞形态和超微变化,ELISA法检测细胞中IL-1β、IL-6、IL-8、TNF-α、NF-κB含量,Western Blot法检测细胞中TLR2、MyD88、NF-κB蛋白表达。结果电镜显示HMGB1组细胞形态和超微结构损伤明显,补肺汤冻干粉和含药血清能有效恢复细胞形态和超微结构损伤,且浓度越高,恢复效果越显著。与空白组比较,HMGB1组细胞中IL-1β、IL-6、IL-8、TNF-α、NF-κB含量均升高(P<0.01),TLR2、MyD88、NF-κB蛋白表达升高(P <0.01);与HMGB1组比较,冻干粉组和含药血清组能浓度依赖性地降低细胞中IL-1β、IL-6、IL-8、TNF-α、NF-κB含量,且0.25%和0.30%冻干粉组及15%、20%和25%含药血清组含量降低(P <0.05,P <0.01);浓度依赖性抑制TLR2、MyD88、NF-κB蛋白表达,且0.30%冻干粉组和25%含药血清组抑制作用明显(P<0.01);补肺汤冻干粉与含药血清各浓度组间各指标比较差异无统计学意义(P>0.05)。结论补肺汤冻干粉及含药血清对HMGB1诱导的A549和HFL1细胞损伤有保护作用,并能减轻炎性因子的释放,机制可能与下调TLR2/MyD88/NF-κB信号通路有关。Objective To investigate the effects of Bufei Decoction(BFD)on morphological damage, inflammatory response, and toll like receptors 2/myeloid differential protein-88/nuclear factor kappa-B(TLR2/MyD88/NF-κB)signaling pathways in human non-small cell lung cancer(A549) cells and human lung fibroblasts(HFL1) cells induced by high mobility family protein B1(HMGB1). Methods BFD lyophilized powder was prepared by freezing technique, the medicated serum was taken after gavage of BFD for 4 weeks. A549 cells and HFL1 cells were randomly divided into the blank group, HMGB1 group(HMGB1-induction), BFD lyophilized powder pre-culture plus HMGB1 group(lyophilized powder group, the concentration of lyophilized powder was 0.10%,0.15%, 0.20%, 0.25%, and 0.30%,respectively), BFD medicated serum pre-culture plus HMGB1 group(medicated serum group,the concentration of medicated serum was 5%, 10%, 15%, 20%, and 25%, respectively), positive drug group(methylprednisolone) and blank serum group,these cells were treated in their respective groups. The cell morphology and ultrastructural changes of each group were observed under electron microscope. The contents of IL-1β, IL-6, IL-8,TNF-α and NF-κB were detected by ELISA,and the expressions of TLR2,MyD88 and NF-κB were detected by Western Blot. Results Electron microscope showed that HMGB1 induced obvious cell morphology and ultrastructure damage were obvious,and lyophilized powder and drug-containing of BFD could effectively restore cell morphology and ultrastructure damage, and the higher the concentration, the more significant the recovery effect. The levels of IL-1β, IL-6, IL-8, TNF-α and NF-κB in HMGB1 induced cells were significantly higher than those in the blank control group(P<0.01),and the protein expressions of TLR2, MyD88 and NF-κB were increased(P<0.01). Compared with HMGB1 group, the contents of IL-1β,IL-6, IL-8,TNF-α and NF-κB in lyophilized powder and medicated serum groups were decreased in a concentration-dependent manner,and the contents in 0.25% and 0.30% lyo

关 键 词：补肺汤 人非小细胞肺癌细胞 人肺成纤维细胞 Toll样受体 转导通路 

分 类 号：R285[医药卫生—中药学]