西瑞香素通过介导巨噬细胞向M2极化对鼻咽癌大鼠线粒体通路的影响  

Effects of daphnoretin on mitochondrial pathway in rats with nasopharyn-geal carcinoma by mediating macrophage polarization to M2

作　　者：王虹园 王继国[1] 任庆[1] WANG Hong-Yuan;WANG Ji-Guo;REN Qing(Department of Otolaryngology,Tianjin Fifth Central Hospital,Tianjin 300450,China)

机构地区：[1]天津市第五中心医院耳鼻咽喉科,天津300450

出　　处：《中国老年学杂志》2023年第20期5013-5018,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(No.81503622)。

摘　　要：目的探究西瑞香素通过介导巨噬细胞向M2极化对鼻咽癌大鼠线粒体通路的影响。方法选取40只SPF级SD雄性大鼠,随机分为正常(N)组、模型(M)组、盐酸千金藤碱(C)组、西瑞香素(W)组,每组10只,对M、C、W组采用腋部皮下注射诱癌剂进行鼻咽癌建模,N组不建立该模型,建模成功后,对C组灌胃15 mg/kg的盐酸千金藤碱,对W组灌胃24 mg/kg的西瑞香素,N组、M组同期给予灌胃同体积生理盐水,将体外培养后的巨噬细胞分为两组,加入磷酸盐缓冲液记为A组,加入西瑞香素记为B组。苏木素-伊红(HE)染色法检测鼻咽组织病理形态,RT-聚合酶链反应(PCR)法检测线粒体通路相关指标,采用末端脱氧核苷酸标记法(TUNEL)检测细胞凋亡,Western印迹法检测肝组织巨噬细胞中CD68、CD163蛋白表达。结果N组鼻咽组织细胞层次清晰且大小均匀,排列整齐;M组细胞层次变多,细胞形态及排列均不整齐,可见多细胞核且染色明显变深,出现大量鳞状上皮增生及炎性细胞浸润,而C、W组症状明显改善,且W组比C组改善明显。与N组比较,M组细胞凋亡明显降低(P<0.05);与M组比较,C、W组细胞凋亡率均明显升高,但与C组相比,W组细胞凋亡率显著升高(P<0.05)。与N组比较,M组鼻咽组织中Bcl-2 mRNA表达显著升高,Bax、Cyto-C mRNA表达显著降低(P<0.05);与M组比较,C、W组鼻咽组织中Bcl-2 mRNA表达显著降低,Bax、Cyto-C mRNA表达明显升高(P<0.05),且W组比C组变化显著。与N组比较,M组鼻咽组织巨噬细胞中CD68蛋白表达显著降低,CD163蛋白表达显著升高(P<0.05);与M组比较,C、W组鼻咽组织巨噬细胞中CD68蛋白表达显著升高,CD163蛋白表达显著降低(P<0.05),且W组比C组变化显著;与A组相比,B组CD68蛋白表达显著增加,CD163蛋白表达显著降低(P<0.05)。结论西瑞香素可影响大鼠线粒体通路表达,促进鼻咽癌细胞凋亡,其作用机制可能与抑制巨噬细胞向M2极化有关。Objective To investigate the effects of daphnoretin on mitochondrial pathway in nasopharyngeal carcinoma rats by mediating macrophage polarization to M2.Methods Forty male SPF grade SD rats were selected and randomly divided into normal(N)group,model(M)group,stephanoline hydrochloride(C)group and daphnoretin(W)group,with 10 rats in each group.Groups M,C and W were injected with subaxillary branch cancer inducer for nasopharyngeal carcinoma modeling.Group N did not establish the model.After successful modeling,group C was given 15 mg/kg stephanine hydrochloride,group W was given 24 mg/kg daphnoretin,groups N and M were given the same volume of normal saline at the same time.Macrophages cultured in vitro were divided in two groups,group A with phosphate buffer and group B with daphnoretin.Hematoxylin-eosin(HE)staining was used to detect the pathological morphology of nasopharynx tissue,and mitochondrial pathway related indexes were detected by RT-polymerase chain reaction(PCR).TdT-mediated dUTP nick end labeling(TUNEL)was used to detect apoptosis,and Western blotting was used to detect the expressions of CD68 and CD163 protein in liver macrophages.Results In group N,the cells of nasopharyngeal tissue had clear layers,uniform size,and orderly arrangement.In group M,the cells had more layers,irregular cell morphology and arrangement,multiple nuclei and staining were visible,and a large number of squamous epithelial hyperplasia and inflammatory cell infiltration appeared,while the symptoms of group C and group W were significantly improved,and the symptoms of group W were significantly improved than group C.Compared with group N,cell apoptosis in group M was significantly decreased(P<0.05);compared with group M,the apoptosis rate in groups C and W was significantly increased,but compared with group C,the apoptosis rate in group W was significantly increased(P<0.05).Compared with group N,the expression of Bcl-2 mRNA in nasopharyngeal tissue of group M was significantly increased,while the expressions of Bax and Cyto-C

关 键 词：西瑞香素 巨噬细胞极化 鼻咽癌 线粒体通路 

分 类 号：R739.63[医药卫生—肿瘤]