芜菁酸性多糖对人肺鳞癌H226细胞自噬的影响  

Effect of acide polysaccharides from Brassica rapa L.(BPA)on autophagy in H226 cells of human lung squamous cell carcinoma

作　　者：李明珠 雅森·米吉提[2] 冯旭瑶 夏腾飞 木巴拉克·伊明江[1] 海力茜·陶尔大洪[1] LI Mingzhu;Yasen Mijiti;FENG Xuyao;XIA Tengfei;Mubalake Yimingjiang;Hailiqian Taoerdahong(College of Pharmacy,Xinjiang Medical University,Urumqi 830017,China;College of International Education,Xinjiang Medical University,Urumqi 830017,China;College of First Clinical Medical,Xinjiang Medical University,Urumqi 830017,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830017 [2]新疆医科大学国际教育学院,乌鲁木齐830017 [3]新疆医科大学第一临床医学院,乌鲁木齐830017

出　　处：《新疆医科大学学报》2025年第4期516-521,529,共7页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区重大科技专项项目(2022A03007-3);新疆天然药物活性组分与释药技术重点实验室项目(XJDX1713)。

摘　　要：目的研究芜菁酸性多糖(Acide polysaccharides from Brassica rapa L.,BPA)对人肺鳞癌细胞系NCI-H226(H226)细胞自噬的影响。方法采用水提醇沉结合色谱技术分离纯化获得两个芜菁酸性多糖(BPA-1和BPA-2)。采用间羟基联苯法测定BPA-1、BPA-2中糖醛酸的含量并对其进行方法学考察。体外培养H226细胞,采用Cell Counting Kit-8(CCK-8)试剂盒检测BPA-1和BPA-2对H226细胞活力的影响。黄嘌呤氧化酶(Xanthine oxidase,XOD)诱导H226细胞建立自噬模型,采用单丹磺酰尸胺(Monodansylcadaverin,MDC)法通过细胞自噬染色检测试剂盒判断细胞自噬模型的建立情况。电镜下观察细胞给药前后自噬形态的变化,活性氧(Reactive oxygen,ROS)试剂盒检测细胞给药前后ROS水平的变化。结果BPA-1和BPA-2中糖醛酸含量分别为48.1%和20.8%。CCK-8结果显示,与空白组相比,BPA-1和BPA-2组的给药浓度达到1000μg/mL时,H226细胞活力明显降低,差异具有统计学意义(P<0.05),且BPA-1和BPA-2的半抑制浓度(Half maximal inhibitory concentration,IC 50)分别为1551μg/mL和6117μg/mL,由此确定BPA-1和BPA-2的低、中和高给药剂量浓度分别为1000、2000、4000μg/mL。MDC法结果显示,与正常组相比,模型组呈强而明显的绿色荧光,且数目较多。电镜结果显示,与梭形、边缘清晰和胞质均匀透亮的正常组细胞相比,模型组细胞圆润、无棱角,胞质内容物被逐渐溶解,出现自噬小体,XOD成功诱导H226细胞自噬模型;与模型组相比,BPA-1低、中剂量组部分细胞可见棱角、部分细胞出现自噬形态,高剂量组大部分细胞呈现出正常形态,且高、中、低剂量组细胞数量均有所减少;BPA-2高剂量组细胞形态趋于正常组,且细胞数目明显减少,中剂量组细胞多呈现自噬形态,细胞数目有所减少,低剂量组的细胞数目和形态均与模型组无明显差异。ROS检测结果显示,与模型组相比,BPA-1和BPA-2低、中、高剂量组ROS含量明显降低,差�Objective To investigate the effect of acide polysaccharides from Brassica rapa L.(BPA)on autophagy in NCI-H226(H226)cells of human lung squamous cell carcinoma.Methods 2 acide polysaccharides from Brassica rapa L.(BPA-1 and BPA-2)were obtained by separating and purifying with the technology of water extraction and alcohol precipitation combined with chromatography.The content of uronic acids in BPA-1 and BPA-2 was determined using the m-hydroxybiphenyl method and methodological investigation was conducted.H226 cells were cultured in vitro,and the effects of BPA-1 and BPA-2 on the viability of H226 cells were detected by Cell Counting Kit-8(CCK-8)kit.Xanthine oxidase(XOD)was used to induce H226 cells to establish an autophagy model,and the establishment of the cell autophagy model was judged by the cell autophagy staining detection kit using the monodansylcverin(MDC)method.The changes of autophagic morphology were observed under an electron microscope and the level of reactive oxygen species(ROS)was detected by the ROS kit before and after drug administration.Results The contents of uronic acids in BPA-1 and BPA-2 were 48.1%and 20.8%,respectively.The results of CCK-8 showed that at the administration concentration of 1000μg/mL,the viabilities of H226 cells in groups of BPA-1 and BPA-2 were significantly decreased compared with the blank group,with a statistically significant difference(P<0.05).And the half maximal inhibitory concentration(IC 50)of BPA-1 and BPA-2 were 1551μg/mL and 6117μg/mL,so the low,medium and high dose concentrations of BPA-1 and BPA-2 were determined as 1000,2000 and 4000μg/mL,respectively.The results of MDC method showed that the model group had stronger green fluorescence and more numbers compared with the normal group.The results of electron microscopy showed that compared with the normal group cells,which were spindle-shaped,with clear edges and uniform and cytoplasm,the model group cells were round and blunt,and the content of the cytoplasm was gradually dissolved,and autophagic bod

关 键 词：人肺鳞癌细胞系 细胞自噬 芜菁酸性多糖 糖醛酸含量 

分 类 号：R917[医药卫生—药物分析学]