Background:Previous studies showed that New Zealand-grown ginseng contains an abundance of ginsenosides and that rare less polar ginsenosides,such as Rg3,exhibit more pharmacological activities than polar ginsenosides...Background:Previous studies showed that New Zealand-grown ginseng contains an abundance of ginsenosides and that rare less polar ginsenosides,such as Rg3,exhibit more pharmacological activities than polar ginsenosides,which are the major components of ginseng.Methods:The ginsenoside profile of New Zealand-grown Panax ginseng was manipulated by treatment with acetic acid,sodium hydroxide,pH,and high temperature.The abundance of 23 ginsenosides extracted by different treatments was quantified using high-performance liquid chromatography.Results:Treatment with 0.5 mol/L acetic acid can stimulate the degradation of polar ginsenosides to less polar ginsenosides(5.6%Rg3 was accumulated,P<0.0001).Furthermore,when ginseng root was treated at 121℃ for 100 min in a pH 3.0 acetic acid aqueous solution,the majority of the polar ginsenosides were converted into less polar ginsenosides.Specifically,83.46±3.69%(P=0.0360)of the less polar ginsenosides and 41.01±2.39%(P=0.0412)of Rg3 were enriched.In contrast,alkali treatment did not convert the polar ginsenosides into less polar ginsenosides at mild temperature and less conversion was observed compared with acid treatment at high temperature.Conclusion:This is the first attempt to manipulate the ginsenoside profile of New Zealand-grown ginseng.The conditions(high temperature with low pH)may be modified to produce and enrich the less polar ginsenoside fraction(especially Rg3)from the total ginseng extract.展开更多
Objective:To determine the effects of ginsenoside rg3 on the body weight of C57BL/6J obese mice and to investigate its underlying weight loss mechanisms with a focus on white fat browning-related factors.Methods:Eight...Objective:To determine the effects of ginsenoside rg3 on the body weight of C57BL/6J obese mice and to investigate its underlying weight loss mechanisms with a focus on white fat browning-related factors.Methods:Eight-week-old C57BL/6J male mice were fed a high-fat diet for 12 successive weeks to construct the obese model.C57BL/6J male mice were fed a standard chow diet to construct normal control group.After 8 weeks of intervention with ginsenoside rg3,the food intake,body weight,body fat mass,blood sugar,and lipid profiles of the mice in each group were detected.Hematoxylin and eosin(HE)staining was used to observe the histological morphology of the adipose tissues.Real-time polymerase chain reaction(RT-PCR)and Western blotting(WB)were applied to detect the gene and protein expression levels of peroxisome proliferators-activated receptor gama(PPARg),Peroxisome proliferatoractivated receptor-gamma coactivator-1alpha(PGC-1a),PR domain containing 16(PRDM16),and uncoupling protein 1(UCP-1).Results:Compared to normal control group mice,the body weight,food intake,body fat composition,and blood lipid levels of model group mice increased significantly.After 8 weeks of intervention with ginsenoside rg3,body weight,body fat composition,food intake,and blood lipid profiles decreased.HE staining showed that ginsenoside rg3 can improve white adipocyte hypertrophy to a certain extent.RTPCR and WB demonstrated that ginsenoside rg3 can increase the mRNA and protein expression levels of PPARg,PGC-1a,PRDM16,and UCP-1 in the adipose tissues of obese mice.Conclusion:The weight reduction effect of ginsenoside rg3 may be related to the promotion of white fat browning.展开更多
Objective:Preeclampsia(PE)is a common complication during pregnancy.miR-100a is expressed in the placenta and regulates the survival and development of placental cells.Insulin growth factor-2(IGF-2)may serve as its do...Objective:Preeclampsia(PE)is a common complication during pregnancy.miR-100a is expressed in the placenta and regulates the survival and development of placental cells.Insulin growth factor-2(IGF-2)may serve as its downstream target.This study investigated the protective mechanisms of ginsenoside Rg3 against PE in rat model.Materials and Methods:LPS-induced rat PE models were suitable for intravenous administration of the highly expressed miR-100a ginsenoside Rg3 lentiviral vector.Human trophoblasts were cultured in vitro for JEG-3,and PE cell models were constructed.In vivo effects on tumor growth and apoptosis were observed.Ginsenoside Rg3 was treated with different concentrations of shRNA,miR-100a analogs,inhibitors,or IGF-2.Autophagy and the expression of autophagy-related proteins were examined.Trophoblast activity and migration were determined using Cell Counting Kit-8 and Transwell assays.Both drugs strongly inhibited trophoblasts under normal conditions with some synergy between them.Double-luciferase return assay confirmed the binding affinity of miR-100a for IGF-2.Results:In response to Rg3,autophagy and the expression of autophagy-related proteins LC3-I/II,Beclin1,and SQSTM1 were reduced in PE rat placental trophoblasts.Rg3 inhibited autophagy in JEG-3 cells and promoted JEG-3 survival and migration in a concentration-dependent manner.miR-100a upregulated PE expression.These results suggested that autophagy was a vital signaling system.Rg3 intervention inhibited miR-100a expression and miR-100a downregulated IGF-2 expression in placental tissues and promoted autophagy,thereby inhibiting JEG-3cell survival and migration.In rats,Rg3 inhibited PE development by regulating the activity of the miR-100a-IGF-2 signaling axis.Conclusion:Ginsenoside Rg3 positively regulates the miR-100a-IGF-2 axis and protects PE rats by inhibiting trophoblastic autophagy and promoting trophoblastic cell survival and migration.展开更多
The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improv...The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improve yields of Rgin a Panax ginseng chassis,combining engineering of triterpene metabolism and overexpression of a lignin biosynthesis gene,phenylalanine ammonia lyase(PAL).We first performed semi-rational design and site mutagenesis to improve the enzymatic efficiency of Pq3-O-UGT2,a glycosyltransferase that directly catalyzes the biosynthesis of Rgfrom Rh.Next,we used clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing to knock down the branch pathway of protopanaxatriol-type ginsenoside biosynthesis to enhance the metabolic flux of the protopanaxadiol-type ginsenoside Rg.Overexpression of PAL accelerated the formation of the xylem structure,significantly improving ginsenoside Rgaccumulation(to 6.19-fold higher than in thecontrol).Wecombinedoverexpression of the ginsenoside aglycon synthetic genes squalene epoxidase,Pq3-O-UGT2,and PAL with CRISPR/Cas9-based knockdown of CYP716A53v2 to improve ginsenoside Rgaccumulation.Finally,we produced ginsenoside Rgat a yield of 83.6 mg/L in a shake flask(7.0 mg/g dry weight,21.12-fold higher than with wild-type cultures).The highproduction system established in this study could be a potential platform to produce the ginsenoside Rgcommercially for pharmaceutical use.展开更多
A murine primary mammary tumor model was established to investigate the treatment with ginsenosides Rg3.The relationship between ginsenosides Rg3 and primary mammary tumor was explored.Mammary tumor was induced by usi...A murine primary mammary tumor model was established to investigate the treatment with ginsenosides Rg3.The relationship between ginsenosides Rg3 and primary mammary tumor was explored.Mammary tumor was induced by using the 7,12-dimethybenz(a)anthracene(DMBA).Ginsenoside Rg3 was employed for treatment.The incidence of mammary tumor in every group was compared,and the expressions of vascular endothelial growth factor(VEGF)and microvessel density(MVD)were detected by immunohistochemical method.The cell cycle and apoptosis percentage were determined by means offlow cytometry.The incidence of tumor in treatment group was significantly lower than that in control group(60.00%vs 33.33%,P<0.05).The average diameter of mammary tumor was(0.86�0.27)cm in control group and(0.39�0.09)cm in treatment group,with the difference being significant between control and treatment groups(P<0.01).The MVD value was(31.9�5.3)in control group and(20.1�4.9)in treatment group,respectively.There was a significant difference between the two groups(P<0.05).The apoptosis percentage in control group was significantly lower than that in treatment group[(2.47�0.69)%vs(5.67�0.99)%,P<0.05].Ginsenoside Rg3 can play an antitumor role in primary mammary tumor model by inhibiting angiogenesis,cell cycle progression,and promoting cell apoptosis.展开更多
文摘Background:Previous studies showed that New Zealand-grown ginseng contains an abundance of ginsenosides and that rare less polar ginsenosides,such as Rg3,exhibit more pharmacological activities than polar ginsenosides,which are the major components of ginseng.Methods:The ginsenoside profile of New Zealand-grown Panax ginseng was manipulated by treatment with acetic acid,sodium hydroxide,pH,and high temperature.The abundance of 23 ginsenosides extracted by different treatments was quantified using high-performance liquid chromatography.Results:Treatment with 0.5 mol/L acetic acid can stimulate the degradation of polar ginsenosides to less polar ginsenosides(5.6%Rg3 was accumulated,P<0.0001).Furthermore,when ginseng root was treated at 121℃ for 100 min in a pH 3.0 acetic acid aqueous solution,the majority of the polar ginsenosides were converted into less polar ginsenosides.Specifically,83.46±3.69%(P=0.0360)of the less polar ginsenosides and 41.01±2.39%(P=0.0412)of Rg3 were enriched.In contrast,alkali treatment did not convert the polar ginsenosides into less polar ginsenosides at mild temperature and less conversion was observed compared with acid treatment at high temperature.Conclusion:This is the first attempt to manipulate the ginsenoside profile of New Zealand-grown ginseng.The conditions(high temperature with low pH)may be modified to produce and enrich the less polar ginsenoside fraction(especially Rg3)from the total ginseng extract.
基金This study received support from the Key Research Project of Beijing University of Chinese Medicine(2020-JYB-ZDGG-029)the National Natural Science Foundation of China(81274041 and 81503540)+1 种基金the Key Drug Development Programme of the Ministry of Science and Technology(20122X09103201-005)the International Cooperation Projects of the Ministry of Education(2011DFA30920).
文摘Objective:To determine the effects of ginsenoside rg3 on the body weight of C57BL/6J obese mice and to investigate its underlying weight loss mechanisms with a focus on white fat browning-related factors.Methods:Eight-week-old C57BL/6J male mice were fed a high-fat diet for 12 successive weeks to construct the obese model.C57BL/6J male mice were fed a standard chow diet to construct normal control group.After 8 weeks of intervention with ginsenoside rg3,the food intake,body weight,body fat mass,blood sugar,and lipid profiles of the mice in each group were detected.Hematoxylin and eosin(HE)staining was used to observe the histological morphology of the adipose tissues.Real-time polymerase chain reaction(RT-PCR)and Western blotting(WB)were applied to detect the gene and protein expression levels of peroxisome proliferators-activated receptor gama(PPARg),Peroxisome proliferatoractivated receptor-gamma coactivator-1alpha(PGC-1a),PR domain containing 16(PRDM16),and uncoupling protein 1(UCP-1).Results:Compared to normal control group mice,the body weight,food intake,body fat composition,and blood lipid levels of model group mice increased significantly.After 8 weeks of intervention with ginsenoside rg3,body weight,body fat composition,food intake,and blood lipid profiles decreased.HE staining showed that ginsenoside rg3 can improve white adipocyte hypertrophy to a certain extent.RTPCR and WB demonstrated that ginsenoside rg3 can increase the mRNA and protein expression levels of PPARg,PGC-1a,PRDM16,and UCP-1 in the adipose tissues of obese mice.Conclusion:The weight reduction effect of ginsenoside rg3 may be related to the promotion of white fat browning.
文摘Objective:Preeclampsia(PE)is a common complication during pregnancy.miR-100a is expressed in the placenta and regulates the survival and development of placental cells.Insulin growth factor-2(IGF-2)may serve as its downstream target.This study investigated the protective mechanisms of ginsenoside Rg3 against PE in rat model.Materials and Methods:LPS-induced rat PE models were suitable for intravenous administration of the highly expressed miR-100a ginsenoside Rg3 lentiviral vector.Human trophoblasts were cultured in vitro for JEG-3,and PE cell models were constructed.In vivo effects on tumor growth and apoptosis were observed.Ginsenoside Rg3 was treated with different concentrations of shRNA,miR-100a analogs,inhibitors,or IGF-2.Autophagy and the expression of autophagy-related proteins were examined.Trophoblast activity and migration were determined using Cell Counting Kit-8 and Transwell assays.Both drugs strongly inhibited trophoblasts under normal conditions with some synergy between them.Double-luciferase return assay confirmed the binding affinity of miR-100a for IGF-2.Results:In response to Rg3,autophagy and the expression of autophagy-related proteins LC3-I/II,Beclin1,and SQSTM1 were reduced in PE rat placental trophoblasts.Rg3 inhibited autophagy in JEG-3 cells and promoted JEG-3 survival and migration in a concentration-dependent manner.miR-100a upregulated PE expression.These results suggested that autophagy was a vital signaling system.Rg3 intervention inhibited miR-100a expression and miR-100a downregulated IGF-2 expression in placental tissues and promoted autophagy,thereby inhibiting JEG-3cell survival and migration.In rats,Rg3 inhibited PE development by regulating the activity of the miR-100a-IGF-2 signaling axis.Conclusion:Ginsenoside Rg3 positively regulates the miR-100a-IGF-2 axis and protects PE rats by inhibiting trophoblastic autophagy and promoting trophoblastic cell survival and migration.
基金financial support from the National Key Research and Development Program(2020YFA0907903)a key project at the central government level:“The ability to establish a sustainable use for valuable Chinese medicine resources”(2060302)+2 种基金the National Science Foundation of China(91954112 and 31900501)the Young Elite Scientists Sponsorship Program of Tianjin(TJSQNTJ-2020-19)the Scientific Research Transformation Foundation of the Wenzhou Safety(Emergency)Institute of Tianjin University。
文摘The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improve yields of Rgin a Panax ginseng chassis,combining engineering of triterpene metabolism and overexpression of a lignin biosynthesis gene,phenylalanine ammonia lyase(PAL).We first performed semi-rational design and site mutagenesis to improve the enzymatic efficiency of Pq3-O-UGT2,a glycosyltransferase that directly catalyzes the biosynthesis of Rgfrom Rh.Next,we used clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing to knock down the branch pathway of protopanaxatriol-type ginsenoside biosynthesis to enhance the metabolic flux of the protopanaxadiol-type ginsenoside Rg.Overexpression of PAL accelerated the formation of the xylem structure,significantly improving ginsenoside Rgaccumulation(to 6.19-fold higher than in thecontrol).Wecombinedoverexpression of the ginsenoside aglycon synthetic genes squalene epoxidase,Pq3-O-UGT2,and PAL with CRISPR/Cas9-based knockdown of CYP716A53v2 to improve ginsenoside Rgaccumulation.Finally,we produced ginsenoside Rgat a yield of 83.6 mg/L in a shake flask(7.0 mg/g dry weight,21.12-fold higher than with wild-type cultures).The highproduction system established in this study could be a potential platform to produce the ginsenoside Rgcommercially for pharmaceutical use.
文摘A murine primary mammary tumor model was established to investigate the treatment with ginsenosides Rg3.The relationship between ginsenosides Rg3 and primary mammary tumor was explored.Mammary tumor was induced by using the 7,12-dimethybenz(a)anthracene(DMBA).Ginsenoside Rg3 was employed for treatment.The incidence of mammary tumor in every group was compared,and the expressions of vascular endothelial growth factor(VEGF)and microvessel density(MVD)were detected by immunohistochemical method.The cell cycle and apoptosis percentage were determined by means offlow cytometry.The incidence of tumor in treatment group was significantly lower than that in control group(60.00%vs 33.33%,P<0.05).The average diameter of mammary tumor was(0.86�0.27)cm in control group and(0.39�0.09)cm in treatment group,with the difference being significant between control and treatment groups(P<0.01).The MVD value was(31.9�5.3)in control group and(20.1�4.9)in treatment group,respectively.There was a significant difference between the two groups(P<0.05).The apoptosis percentage in control group was significantly lower than that in treatment group[(2.47�0.69)%vs(5.67�0.99)%,P<0.05].Ginsenoside Rg3 can play an antitumor role in primary mammary tumor model by inhibiting angiogenesis,cell cycle progression,and promoting cell apoptosis.
