首页|期刊导航|山东医药|装载人参皂苷Rh2外泌体的制备及其对肝癌细胞株Huh7、PLC半数抑制浓度测算

装载人参皂苷Rh2外泌体的制备及其对肝癌细胞株Huh7、PLC半数抑制浓度测算OACSTPCD

Preparation of ginsenoside Rh2-loaded exosomes and estimation of IC50 on hepatoma cell lines Huh7 and PLC

中文摘要英文摘要

目的 制备装载人参皂苷Rh2(G-Rh2)的外泌体(Exos@G-Rh2),并测算其对肝癌细胞株Huh7、PLC的半数抑制浓度(IC50).方法 利用外泌体提取试剂盒从肝癌Huh7细胞培养上清中分离提纯外泌体(Exos),采用电穿孔法制备装载G-Rh2的外泌体Exos@G-Rh2,使用透射电子显微镜观察Exos@G-Rh2、Exos形态,使用纳米颗粒跟踪分析仪测算Exos@G-Rh2、Exos粒径,采用高效液相色谱(HPLC)法测算Exos@G-Rh2中G-Rh2含量,并计算Exos@G-Rh2载药量.采用PKH-26荧光染色法观察肝癌细胞株Huh7、PLC对Exos@G-Rh2的摄取情况,采用CCK-8法测算Exos@G-Rh2、游离G-Rh2对Huh7细胞、PLC细胞的IC50.结果 Exos及Exos@G-Rh2在电镜下均可见经典的茶托状结构,且有质膜包裹,形态无显著差异.Exos@G-Rh2、Exos粒径分别为(156.90±10.23)nm、(143.47±8.62)nm,两者相比,P>0.05.Exos@G-Rh2载药量为24.25%±0.27%.在Huh7细胞、PLC细胞中,红色荧光清晰可见,主要在细胞质中.在Huh7细胞中,游离G-Rh2的IC50值为(34.96±1.37)µg/mL,而Exos@G-Rh2的IC50值为(29.74±2.83)µg/mL,两者相比,P<0.05;在PLC细胞中,游离G-Rh2的IC50值为(33.41±1.47)µg/mL,而Exos@G-Rh2的IC50值为(30.08±1.12)µg/mL,两者相比,P<0.05.结论 成功制备Exos@G-Rh2,其形态和粒径与Exos无显著差异,可被Huh7细胞、PLC细胞摄取.与游离G-Rh2相比,Exos@G-Rh2对Huh7细胞、PLC细胞的IC50更低.

Objective To prepare the exosomes loaded with ginsenoside Rh2(Exos@G-Rh2),and to calculate the half-maximal inhibitory concentration(IC50)on hepatoma cell lines Huh7 and PLC.Methods Using exosome extraction kits to isolate and purify exosomes(Exos)from the culture supernatant of hepatoma Huh7 cells,the electroporation method was employed to prepare Exos@G-Rh2.Transmission electron microscopy was used to observe the morphology of Exos@G-Rh2 and Exos.The particle size of Exos@G-Rh2 and Exos was measured using nanoparticle tracking analysis.High-perfor-mance liquid chromatography(HPLC)was utilized to determine the G-Rh2 content in Exos@G-Rh2,and the drug-loading capacity was calculated.The uptake of Exos@G-Rh2 by hepatoma cell lines Huh7 and PLC was observed using PKH-26 flu-orescence staining.The IC50 of Exos@G-Rh2 and free G-Rh2 on Huh7 and PLC cells was measured using CCK-8.Re-sults Under the transmission electron microscope,both Exos and Exos@G-Rh2 displayed the classic cup-shaped struc-ture enveloped by a lipid membrane,with no significant morphological differences.The particle sizes of Exos@G-Rh2 and Exos were(156.90±10.23)nm and(143.47±8.62)nm,respectively,with no significant difference between them(P>0.05).The drug-loading capacity of Exos@G-Rh2 was 24.25%±0.27%.In the cytoplasm of Huh7 and PLC cells,red fluo-rescence was clearly visible,primarily within the cytoplasm.In the Huh7 cells,the IC50 value of free G-Rh2 was(34.96±1.37)µg/mL,whereas the IC50 value of Exos@G-Rh2 was(29.74±2.83)µg/mL,showing a significant difference(P<0.05).In the PLC cells,the IC50 value of free G-Rh2 was(33.41±1.47)µg/mL,while the IC50 value of Exos@G-Rh2 was(30.08±1.12)µg/mL,also showing a significant difference(P<0.05).Conclusions Exos@G-Rh2 is successfully prepared,with its morphology and particle size showing no significant difference from Exos,and it can be taken up by Huh7 and PLC cells.Compared with free G-Rh2,Exos@G-Rh2 exhibits a lower IC50 for Huh7 and PLC cells.

李美乐;赵梓粤;金凯;唐婷;黄俊期;王戈睿;谢裕安

广西医科大学附属肿瘤医院实验研究部,南宁 530021广西医科大学第一附属医院消化内科广西中医药大学研究生院广西医科大学附属肿瘤医院实验研究部,南宁 530021||广西中医药大学研究生院||广西壮族自治区妇幼保健院

临床医学

人参人参皂苷Rh2外泌体肝细胞癌药物载体

ginsengginsenoside Rh2exosomeshepatocellular carcinomadrug delivery

《山东医药》 2024 (009)

33-36 / 4

广西重点实验室运行补助项目(21-220-22).

10.3969/j.issn.1002-266X.2024.09.008

评论