化痰祛湿活血方对脂多糖诱导的巨噬细胞焦亡形态的影响  

Effect of Huatan Qushi Huoxue prescription on lipopolysaccharide-induced pyroptosis of macrophages

作　　者：刘鸣昊[1] 刘素彤 尚东方[1] 张丽慧[1] 顾亚娇[1] 赵文霞[1] LIU Minghao;LIU Sutong;SHANG Dongfang;ZHANG Lihui;GU Yajiao;ZHAO Wenxia(Department of Hepatology and Spleen-Stomach,The First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院脾胃肝胆科,郑州450000

出　　处：《临床肝胆病杂志》2022年第9期2016-2019,共4页Journal of Clinical Hepatology

基　　金：国家自然科学基金项目(81904154);河南省科技攻关计划(202102310168);河南省重点研发与推广专项课题(192102310425);河南省中医药科学研究专项课题(2019JDZX2051);河南省科技攻关计划项目(202102310495);河南省特色骨干学科中医学学科建设项目(STG-ZYXKY-2020024)。

摘　　要：目的化痰祛湿活血方对脂多糖(LPS)诱导的RAW264.7细胞焦亡形态的影响及机制研究。方法复制体外LPS诱导活化RAW264.7细胞模型,分为空白组,模型组,化痰祛湿活血方高、中、低剂量组,对照组。经相应的含药血清干预24 h。扫描电子显微镜观察各组细胞形态。免疫荧光对GSDMD-N进行定量定位分析。结果空白组细胞形态圆形规整,表面光滑;对照组细胞肿胀,表面有褶皱,包膜有隙,符合细胞焦亡形态。对照组细胞表面有气泡状突出物,伪足明显。细胞膜有孔隙,高剂量组细胞形态无肿胀,表面粗糙有伪足,胞膜上未见明显孔隙。低剂量和中剂量组细胞肿胀,细胞膜表面粗糙有孔隙,有伪足伸出。免疫荧光结果显示,与空白组相比模型组GSDMD-N阳性染色强度明显增加,对照组与中药组较模型组GSDMD-N阳性染色强度均有减弱。结论化痰祛湿活血方可改善巨噬细胞焦亡现象,减少GSDMD-N表达。Objective To investigate the effect of Huatan Qushi Huoxue prescription on lipopolysaccharide(LPS)-induced pyroptosis of RAW264.7 cells and its mechanism.Methods An in vitro cell model of LPS-induced activated RAW264.7 was established and divided into blank group,model group,high-,middle-,and low-dose Huatan Qushi Huoxue prescription groups,and control group.The corresponding drug-containing serum intervention was performed for 24 hours.A scanning electron microscope was used to observe cell morphology,and immunofluorescence assay was used to perform quantitative localization of GSDMD-N.Results The cells in the blank group were round and regular in shape with smooth surface,and those in the control group were swollen,with folds on the surface and gaps in the capsule,which were consistent with the morphology of cell pyroptosis.The cells in the control group had bubbles on the surface with obvious pseudopodia and pores in cell membrane,and those in the high-dose group were not swollen and had a rough surface with pseudopodia,with no obvious pores in cell membrane.The cells in the low-and middle-dose groups were swollen and had a rough surface of cell membrane with pores and pseudopodia.Immunofluorescence assay showed that compared with the blank group,the model group had a significant increase in the positive staining intensity of GSDMD-N,and compared with the model group,the control group and the Traditional Chinese medicine group had a reduction in the positive staining intensity of GSDMD-N.Conclusion Huatan Qushi Huoxue prescription can improve the pyroptosis of macrophages and reduce the expression of GSDMD-N.

关 键 词：脂肪肝 巨噬细胞 细胞焦亡 

分 类 号：R285[医药卫生—中药学]