Characterization Of Ginsenosides Carbon Nanodots And Their Inhibitory Effect On Neuroblastoma

Objective: To obtain different ginsenosides carbon nanodots(GS-CDs)under different hydrothermal reaction conditions by selecting the main effective component of ginseng as raw material.Study the change rule of its physical and chemical properties,and explore the possible new biological activities and medicinal effects,in order to get ginseng drug products with fluorescent labeling function and certain biological activities.Methods: 1.Preparation of ginsenosides carbon nanodots: take ginsenosides powder as the carbon source,the reaction concentration is 1.5 mg/m L,control the reaction time(1-10 h)at 170 ℃,and obtain different GS-CDs solutions by hydrothermal synthesis.After the solution was coarsely filtered with a 0.22 μm filter membrane,it was purified by centrifugation with an ultrafiltration tube to obtain a pale yellow GS-CDs solution.2.Property analysis of ginsenosides carbon nanodots: the physicochemical properties and structure of GS-CDs were analyzed and characterized by high-resolution transmission electron microscopy(TEM),ultraviolet visible absorption spectroscopy(UV-vis),fluorescence spectroscopy(FL),Fourier transform infrared spectroscopy(FTIR)and X-ray photoelectron spectroscopy(XPS).3.Study on the biological activity of ginsenosides carbon nanodots in vitro: cell counting kit-8 method was used to explore the bioactivity of GS-CDs against different kinds of tumor cells,including rat adrenal pheochromocytoma cells(PC12),human cervical cancer cells(He La),human liver cancer cells(Hep G2),mouse small glial cells(BV2)and human neuroblastoma cells(SH-SY5Y);Fluorescence imaging technology was used to capture fluorescence imaging images of cells incubated with GS-CDs for different times;Apoptosis kit and flow cytometry were used to detect the apoptosis of GS-CDs.4.Study on the biological activity of ginsenosides carbon nanodots in vivo: according to the above experimental results,GS-CDs has obvious inhibitory effect on SH-SY5 Y cells,so the corresponding neuroblastoma tumor bearing mouse model is established.After screening,they were randomly divided into three groups: control group,cisplatin group and administration group.The weight of the mice and the growth of the tumor was recorded and monitored.HE staining was used to detect the pathological changes of the tumors and the major organs of the mice;The expression of related proteins was detected by immunohistochemistry to investigate the inhibitory effect of GS-CDs on neuroblastoma.Results: 1.GS-CDs is quasi spherical,with good dispersion,uniform size,particle size distribution between 2.0 ～ 4.0 nm and strong blue-green fluorescence.With the increase of reaction time,the UV absorption and fluorescence emission of GS-CDs gradually increased.2.FTIR spectrum analysis shows that the main structure of tetracyclic triterpenoids of GS-CDs has little change,and the surface structure is rich in certain hydroxyl and carbonyl structures,which is conducive to its subsequent effective interaction with organisms through functional groups.3.In vitro bioactivity studies showed that GS-CDs had little inhibitory effect on He La,Hep G2,BV2 and PC12 cells,but had strong inhibitory effect on SH-SY5 Y cells and could be ingested by SH-SY5 Y cells.With the increase of administration concentration,the total apoptosis rate of SH-SY5 Y cells increased significantly.4.Through the summary and analysis of the above results,it is concluded that the best reaction condition of GS-CDs is 170 ℃-6 h.The GS-CDs solution synthesized under this condition was administered to mice by tail vein injection.Compared with the model group and cisplatin group,GS-CDS can significantly inhibit the growth of neuroblastoma.And GS-CDs have low toxicity and side effects,and will not cause damage to the body of healthy mice.Conclusion: Under certain conditions,ginsenosides carbon nanodots prepared by hydrothermal synthesis can form nano-aggregates with obvious lattice,rich surface groups,good fluorescence properties,water solubility and biocompatibility.Ginsenosides carbon nanodots show obvious biological activities in vitro and in vivo,significantly inhibited both SH-SY5 Y cells and neuroblastoma,and could be taken up by cells as an effective cellular fluorescent probe.This study provides a new method for the clinical treatment of neuroblastoma,and also provides a good research basis and application prospect for the subsequent preparation and development of nano-drugs with active ingredients of Chinese herbal medicine.