Systematic analysis of the material basis and mechanism of total saponins of mountain cultivated ginseng against doxorubicin-induced cardiotoxicity based on integrating network pharmacology and in vivo substance profiling.

INTRODUCTION: Doxorubicin-induced cardiotoxicity (DIC) is a serious obstacle to oncologic treatment. Mountain cultivated ginseng (MCG) exhibits stronger pharmacological effects than cultivated ginseng (CG) mainly due to the differences in ginsenosides. However, the material basis and the underlying mechanism of the protective effects of total saponins of MCG (TSMCG) against DIC are unclear. OBJECTIVES: We aimed to elucidate the material basis and the pharmacodynamic effects of TSMCG on DIC as well as the underlying mechanisms. METHODS: To comprehensively analyze the effective substances, the chemical components of TSMCG and their prototypes or metabolites in vivo were characterized through UHPLC/Q-TOF-MS. Then, an absorbed component-target-disease network was established to explore the mechanisms underlying the protective effects of TSMCG against DIC. H9c2 cells were employed for pharmacodynamic assays. The mechanism was verified by Western blot and molecular docking simulations. RESULTS: A total of 56 main ginsenosides were identified in TSMCG, including 27 ginsenosides of PPD type, 15 ginsenosides of PPT type, two ginsenosides of OA types, and 12 ginsenosides of other types. Moreover, 55 ginsenoside prototypes or metabolites in vivo were tentatively characterized. Ginsenoside Ra1 , a differential compound between MCG and CG, could be metabolized by oxidation and deglycosylation. Network pharmacology showed that AKT1, p53, and STAT3 are core targets of 62 intersecting genes. Molecular docking results indicated that most of the ginsenosides have favorable affinity with these core targets. After doxorubicin exposure, TSMCG could increase cell viability and inhibit apoptosis in a dose-dependent manner. CONCLUSION: Our work reveals a novel comprehensive strategy to study the material basis of the protective effects of TSMCG against DIC and the underlying mechanisms through integrating in vivo substance identification, metabolic profiling, network pharmacology, pharmacodynamic evaluation, and mechanism verification.

A glycoprotein from mountain cultivated ginseng: Insights into their chemical characteristics and intracellular antioxidant activity.

A glycoprotein (MGP2) from mountain-cultivated ginseng (MCG) was purified by Tris-HCl extraction followed by DEAE-52 ion exchange chromatography and Sephadex G-100 gel filtration chromatography. The approximate molecular weight (27.0 kDa) and monomeric nature were determined by reduced and non-reduced SDS-PAGE. The structure of MGP2 was characterized by a practical and reliable "protein-polysaccharide analyzed by spectroscopy combined with chemical analysis" strategy. The results showed that MGP2 belonged to Arabinogalactan proteins (AGPs) which contained high amount of Glc (35.1 %). The hemagglutination test concluded that MGP2 was not a lectin. In addition, the MGP2 exhibited antioxidant activity by scavenging radical capacity tests and the ability to protect human erythrocytes and RAW264.7 cells from oxidative damage induced by AAPH. Therefore, these results suggested that glycoprotein MGP2 could be used as a natural antioxidant in drug and food industry.

Anthocyanins inhibit high glucose-induced renal tubular cell apoptosis caused by oxidative stress in db/db mice.

Oxidative stress is an important contributory factor resulting the development of kidney injury in patients with diabetes. Numerous in vitro and in vivo studies have suggested that anthocyanins, natural phenols commonly existing in numerous fruits and vegetables, exhibit important antioxidative, anti­inflammatory and antihyperlipidemic effects; however, their effects and underlying mechanisms on diabetic nephropathy (DN) have not yet been fully determined. In the present study, the regulation of apoptosis metabolism and antioxidative effects exhibited by anthocyanins [grape seed procyanidin (GSPE) and cyanidin­3­O­ß­glucoside chloride (C3G)] were investigated, and the molecular mechanism underlying this process was investigated in vivo and in vitro. GSPE administration was revealed to suppress renal cell apoptosis, as well as suppress the expression of Bcl­2 in diabetic mouse kidneys. Furthermore, GSPE administration was demonstrated to suppress the expression of thioredoxin interacting protein (TXNIP), in addition to enhancing p38 mitogen­activation protein kinase (MAPK) and extracellular signal­regulated kinase 1/2 (ERK1/2) oxidase activity in diabetic mouse kidneys. In vitro experiments using HK­2 cells revealed that C3G suppressed the generation of HG­mediated reactive oxygen species, cellular apoptosis, the expression of cleaved caspase­3 and the Bax/Bcl­2 ratio; and enhanced the expression of cytochrome c released from mitochondria. Furthermore, treatment with C3G was revealed to suppress the expression of TXNIP, in addition to the phosphorylation of p38 MAPK and ERK1/2 oxidase activity in HK­2 cells under HG conditions. In addition, treatment with C3G was revealed to attenuate the HG­induced suppression of the biological activity of thioredoxin, and to enhance the expression of thioredoxin 2 in HK­2 cells under HG conditions. In conclusion, the present study demonstrated that anthocyanins may exhibit protective effects against HG­induced renal injury in DN via antioxidant activity.