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