Preliminary Identification of Red Pigment and Positive Correlation between the Contents of Red Pigment and Total Saponins of Panax notoginseng Fruits

[Objective] This study aimed to identify red pigment of Panax notoginseng fruits and explore the correlation between pigment content and total saponins of the fruits. [Method] The red pigment of Panax notoginseng fruits was preliminarily identi- fied with specific color reactions and UV-vis spectra, and the contents of the pigment and total saponins were determined via spectrophotometry. [Result] The red hues of the fruits were contributed by anthocyanins and/or the anthocyanidins. The contents of anthocyanins and total saponins of the fruits both decreased along with thinning of the red hues. The content difference of the anthocyanins in fruits with different red hues reached extremely significant level, but that of total saponins just reached significant level. [Conclusion] The red pigment of P. notoginseng fruits is anthocyanins which are of extremely significant positive correlation with total saponins in contents.

Responses of Photosynthetic Pigment, Phenol and Total Saponin Contents of Panax notoginseng to Supplemental UV-B under Field Conditions

[Objective] The aim of this study was to investigate the content changes and their correlations of the photosynthetic pigment,phenols,including total phenols,total flavonoids and anthocyanins,and total saponins of the one-year-old P.notoginseng plants under supplemental UV-B stress in fields.[Method] The one-year-old plants were irradiated by UV-B in field for 1 min per day,and the plants under the UV-B lamp were regarded as a circle center,achieving an annular leaf-sampling.The photosynthetic pigment,phenols and total saponins of the leaves were determined spectrophotometrically.[Result] With the increase of sampling radius,the supplemental UV-B intensity decreased significantly,the contents of chlorophyll （Chl） a,Chl b,Chl （a＋b）,carotenoid （Car） and total photosynthetic pigment （Chl＋Car） of the leaves increased extremely significantly,the Chl a/b and total phenol content （TPC） decreased extremely significantly,but the Chl （a＋b）/Car changes were not significant.The contents of total flavonoids,anthocyanins and saponins all increased due to the increasing of UV-B,displaying dose effects.The UV-B intensity was positively correlated with the Chl a/b,and negatively with the Chl a,Chl b,Chl （a＋ b）,Car and （Chl＋Car） contents; and the two of TPC,total flavonoid content （TFC）,total anthocyanin content （TAC） and total saponin content （TSC） were positively correlated,all reaching extremely significant level.The UV-B intensity was positively and significantly correlated with the total flavonoid content （TFC）,negatively and significantly with the Chl （a＋b）/Car,and positively and insignificantly with the TPC,TAC and TSC.[Conclusion] For one-year-old plants of P.notoginseng,UV-B can decrease the contents of the Chl a,Chl b,Chl （a＋b）,Car and （Chl＋Car） and increase the Chl a/b and TPC,and,furthermore,induce the increases of the TFC,TAC and TSC in a dose-dependent manner.However,UV-B can hardly change the Chl （a＋b）/Car.The supplemental UV-B of well-suited dose might be one of the effective measures to improve the TSC of P.notoginseng.

Effects of total saponins of Panax notoginseng on immature neuroblasts in the adult olfactory bulb following global cerebral ischemia/reperfusion

The main active components extracted from Panax notoginseng are total saponins. They have been shown to inhibit platelet aggregation, increase cerebral blood flow, improve neurological behavior, decrease infarct volume and promote proliferation and differentiation of neural stem cells in the hippocampus and lateral ventricles. However, there is a lack of studies on whether total saponins of Panax notoginsertg have potential benefits on immature neuroblasts in the olfactory bulb following ischemia and reperfusion. This study established a rat model of global cerebral ischemia and reperfusion using four-vessel occlusion. Rats were administered total sa- ponins of Panax notoginseng at 75 mg/kg intraperitoneally 30 minutes after ischemia then once a day, for either 7 or 14 days. Total saponins of Panax notoginseng enhanced the number of dou- blecortin （DCX）＋ neural progenitor ceils and increased co-localization of DCX with neuronal nuclei and phosphorylated cAMP response element-binding/DCX＋ neural progenitor cells in the olfactory bulb at 7 and 14 days post ischemia. These findings indicate that following global brain ischemia/reperfusion, total saponins of Panax notoginseng promote differentiation of DCX＋ cells expressing immature neuroblasts in the olfactory bulb and the underlying mechanism is related to the activation of the signaling pathway of cyclic adenosine monophosphate response element binding protein.

Neuroprotective effects of total saponins from Rubus parvifolius L. on cerebral ischemia/reperfusion injury in rats

This study examines the neuroprotective effects and mechanisms of action of total saponins from Rubus parvifolius L. （TSRP） on focal cerebral ischemia and reperfusion injury in rats. Focal cerebral ischemia and reperfusion injury was performed in rats using the suture method. The results indicate that intragastric injection of TSRP, at 5, 10 and 20 mg/kg, could decrease neurological impairment, reduce cerebral infarct volume, diminish pathological changes, and significantly inhibit the apoptosis of neurons surrounding the ischemic area. In addition, TSRP upregulated the expression of the anti-apoptotic factor Bcl-2, at the protein and mRNA levels, and it downregulated the expression of the pro-apoptotic factor Bax, at the protein and mRNA levels. These findings indicate that TSRP protects against cerebral ischemia/reperfusion injury, and that it may do so by regulating the expression of Bcl-2 and Bax.

Effects of total soy saponins on free radicals in the quadriceps femoris,serum testosterone,LDH,and BUN of exhausted rats

Purpose:The aim of this study was to investigate the impact of total soy saponins(TS) on the free radical metabolism from the quadriceps femoris muscle,serum testosterone,lactate dehydrogenase(LDH),and blood urea nitrogen(BUN) in rats exercised to exhaustion.Methods:A one-time exhausted treadmill exercise session was used.Sprague-Dawley rats were divided into 4 groups:a control group—animals receiving no TS and no exercise(NTSNE),animals receiving TS but no exercise group(TSNE),animals receiving no TS but exercised to exhaustion group(NTSE),and animals receiving TS and exercised to exhaustion group(TSE).The TSNE and TSE groups were fed TS at a dosage of 20 mg/kg body weight once per day for 2 weeks.The NTSE group was given a placebo,and the NTSNE group was not given any treatment.The NTSE and TSE groups were exercised at speed of 30 m/min on treadmill until exhausted.The exercise time and exercise distance were recorded when the rats became exhausted and the rats were then decapitated and anatomized immediately.A 10% homogenate of the quadriceps femoris tissue was prepared.The levels of superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),glutathione peroxidase(GSH-Px),glutathione reductase(GR),reduced glutathione(GSH),total antioxidant capacity(T-AOC),LDH,BUN,and serum testosterone were tested.Results:TS significantly increased the exercise time to exhaustion by 20.62%(p < 0.05).The MDA levels were decreased significantly in the TSNE group than in NTSNE group(p < 0.05);the T-AOC levels increased significantly in the TSNE group than in the other 3 groups(p < 0.01,p < 0.05,p < 0.05).The LDH activity significantly increased in the NTSE group than in TSNE group(p < 0.05).The BUN levels significantly increased in the NTSE group than in the other 3 groups(p < 0.01,p < 0.01,p < 0.05),and significantly increased in the TSE group than in NTSNE and TSNE groups(both p < 0.01).The serum testosterone levels increased significantly in the TSNE group than in the other 3 groups(all p < 0.01).SOD,CAT,GSH-Px,GR,and GSH were not statistically different among the groups.Conclusion:TS can significantly improve the exercised rats' serum testosterone level and antioxidant activity in their quadriceps femoris to varying degrees,decrease MDA and serum LDH and BUN levels,increase the exercise time,and delay the occurrence of the fatigue.

Total saponins of Panax ginseng effects on proliferation and differentiation of human embryonic neural stem cells and in a Parkinson's disease mouse model

BACKGROUND： Total saponins of Panax ginseng （TSPG） exhibits neuroprotection against Parkinson＇s disease in the substantia nigra. OBJECTIVE： To investigate the effects of TSPG on human embryonic neural stem cells （NSCs） proliferation and differentiation into dopaminergic neurons using in vitro studies, and to observe NSC differentiation in a mouse model of Parkinson＇s disease, as well as behavioral changes before and after transplantation. DESIGN, TIME AND SETTING： In vitro neural cell biology trial and in vivo randomized, controlled animal trial were performed at the Institute of Basic Medical Sciences, Chongqing Medical University between September 2004 and December 2007. MATERIALS： TSPG （purity 〉 95%） was isolated, extracted, and identified by Chongqing Academy of Chinese Materia Medica. Recombinant human basic fibroblast growth factor （bFGF） and recombinant human epidermal growth factor （EGF） were purchased from PeproTech, USA. A total of 25 C57/BL6J mice, aged 18-20 weeks were included. Twenty were used to establish a Parkinson＇s disease model with i.p. injection of MPTP （1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine） and TSPG alone or combined with interleukin-1 （IL-1）-treated NSCs prior to transplantation into the corpus striatum. The remaining five mice were pretreated for 3 days with TSPG prior to MPTP injection, serving as the TSPG prevention group. METHODS： Primary NSCs were isolated, cultured and purified from embryonic cerebral cortex. Immunocytochemistry was employed to detect specific antigen expression in the NSCs. In vitro experiment： （1） to induce proliferation, NSCs were treated with TSPG, EGF＋bFGF, or TSPG＋EGF＋bFGF, respectively; （2） to induce dopaminergic neuronal differentiation, NSCs were treated with TSPG, IL-1, or TSPG＋IL-1, respectively. MAIN OUTCOME MEASURES： In vitro experiment： the effects of TSPG on NSCs proliferation were evaluated with flow cytometry and MTT assay. Tyrosine hydroxylase expression was determined by immunocytochemistry assay to observe effects of TSPG on dopaminergic neuronal differentiation. In vivo experiment： differentiation of grafted NSCs in the mouse brain was determined by immunohistochemical staining. Behavioral changes were evaluated by spontaneous activity frequency, memory function, and score of paralysis agitans. RESULTS： （1） NSCs were cultured and passaged for more than three passages. Immunocytochemistry revealed positive nestin staining, as well as neurofilament protein and glial fibrillary acidic protein. （2） TSPG significantly increased NSC proliferation, in particular when combined with EGF and bFGF, which was twice as effective as FGF or bFGF alone. TSPG also induced dopaminergic differentiation in NSCs, in particular when TSPG was added together with IL-1, resulting in an effect five times greater than that of IL-1 alone. （3） At day 30 following transplantation, most NSCs in the TSPG prevention group differentiated into dopaminergic neurons, and the scores of paralysis agitans, spontaneous activity, and memory function were significantly increased compared with TSPG alone or TSPG＋IL-1 groups （P 〈 0.05）. CONCLUSION： TSPG stimulated NSC proliferation, in particular when combined with FGF and bFGF. TSPG significantly induced dopaminergic neuronal differentiation of NSCs, and the effect was greater when combined with IL-1. In addition, TSPG greatly improved behavior in the Parkinson＇s disease mouse model following NSC transplantation. Following NSC transplantation, TSPG pretreatment exhibited superior efficacy over either TSPG alone or TSPG in combination with IL-1, in terms of behavioral improvements in the Parkinson＇s disease mouse model.

Total soy saponins improve the antioxidant capacity of the myocardium and exercise ability in exhausted rats

Purpose：The aim of the present study was to investigate the impact of total soy saponins（TS） on the myocardial antioxidant capacity in rats exercised to exhaustion.Methods：The one-time exhausted treadmill model was used.All rats were divided into 4 groups：the control group,the TS group,the exhausted group,and the TS exhausted group.The TS and TS exhausted groups were fed TS at a dosage of 20 mg/kg body weight,once a day,for 2 weeks.The exhausted group was given a placebo,and the control group was not given any treatment.The treadmill speed was set at 30 m/min,and the rats（exhausted and TS exhausted groups） were trained at this speed until exhausted.The rats were decapitated and anatomized immediately after exhausted.A 10% homogenate of the myocardial tissue was prepared.Results：TS significant y increased the exercise time by 20.62%（p〈0.05）.As compared with the control group,the enzyme activities for catalase（CAT）,glutathione peroxidase（GSH-Px）,and glutathione reductase（GR） were significant y enhanced in the TS group（p〈0.01）;GR and GSH-Px activity was significant y enhanced in the TS exhausted group（p〈0.01）;malondialdehyde（MDA） levels were significant y decreased in the TS exhausted group（p〈0.05）.As compared with the exhausted group,the GSH-Px activity was significant y enhanced in the TS exhausted group（p〈0.01）;CAT,GSH-Px,and GR activities were significant y enhanced in the TS group（p〈0.01）.As compared with the TS group,the CAT and GR activity in the TS exhausted group was significant y decreased（p〈0.01）.Conclusion：TS can improve the exercised rats＇ antioxidant activity in their cardiac muscle to varying degrees,decrease MDA and serum AST and LDH levels,increase the exercise time,and delay the occurrence of sports fatigue.

THE PROTECTIVE EFFECTS OF THE TOTAL SAPONIN OF DIPSACUS ASPEROIDES ON THE APOPTOSIS OF HIPPOCAMPAL NEURONS INDUCED BY β-AMYLOID PROTEIN

Objective To investigate the effects of the total saponin of Dipsacus asperoides (tSDA) and ginsenoside Rb1 (GRb1) on the apoptosis of primary cultured hippocampal neurons induced by β-amyloid protein (Aβ). Methods Primary cultured hippocampal neurons, the cultures were pretreated with tSDA and GRb1 on 10d for 24 hours respectively. Then the cultures were treated with 35 μmol·L -1 Aβ25-35 for 24 hours, observed the changing of survival rate of neurons and the apoptosis of neurons with biochemical analysis combining immunofluorescent cytochemical double-staining technique. Results Hippocampal neurons were treated with 35 μmol·L -1 Aβ for 24 hours, and survival rate of neurons downed to 52.6%. When neurons were pretreated by tSDA and GRb1, survival rate of neurons increased 11% to 15%. The findings of immunofluorescent cytochemical double-staining indicated that apoptotic neurons were obviously more than that of the blank group, reaching 43.9%.When neurons were pretreated by tSDA and GRb1, apoptotic neurons were downed to 16.6%, 10.8% respectively. Conclusion tSDA had the same effects as GRb1, protecting the neurons, antagonizing neurotoxicity of Aβ, increasing survival rate of neurons, and reducing apoptotic neurons induced by Aβ.

Screening and functional evaluation of the glucose-lowering active compounds of total saponins of Baibiandou(Lablab Semen Album)

Objective To screen forα-glucosidase inhibitor active compounds in the total saponins of Baibiandou(Lablab Semen Album)based on UHPLC-Q-Exactive Orbitrap MS technology and to evaluate its hypoglycemic activity in vivo.Methods Acarbose was used as the positive control,and the median inhibitory concentration(IC50)was used as the evaluation index ofα-glucosidase inhibitory activity to establish an in vitroα-glucosidase inhibition model.Further,UHPLC-Q-Exactive Orbitrap MS technology was used to screen and identify the active compounds ofα-glucosidase inhibitors in the total saponins of Baibiandou(Lablab Semen Album)in order to further verify the activity of the main active monomer and to perform homologous modeling and molecular docking of yeast-derivedα-glucosidase and human-derivedα-glucosidase,while the hypoglycemic activity was evaluated in diabetic mice.Results This study successfully identified 15 compounds with potentialα-glucosidase inhibitory activity,including Chikusetsusaponin IVa,from the total saponins of Baibiandou(Lablab Semen Album).Simultaneously,we verified the activity of the main active monomer Chikusetsusaponin IVa,and showed that it has strongα-glucosidase inhibitory activity.Theα-glucosidase inhibitory concentration IC50 was(565.2±1.026)μg/m L,and the IC50 of acarbose,which was lower than the positive control,was(706.6±1.058)μg/m L.The docking energies of Chikusetsusaponin IVa were–6.1 and–7.7 kcal/mol with yeast-derivedα-glucosidase and human-derivedα-glucosidase molecules,respectively.Both showed strong binding activity,and the levels of alanine aminotransaminase(ALT),aspartate aminotransaminase(AST),UREA,Creatinine(CREA),and cholesterol(CHO)were significantly decreased by Chikusetsusaponin IVa(P<0.05).In addition,it could repair damaged liver and pancreas cells of diabetic mice to some extent.Conclusion This study provides a basis for screeningα-glucosidase inhibitors and structural modifications of the total saponins of Baibiandou(Lablab Semen Album).

Neuroprotection of total steroid saponins from Dioscorea zingiberensis C.H.Wright against transient focal cerebral ischemia-reperfusion injury in rats via its anti-inflammatory and anti-apoptotic effects

OBJECTIVE The total steroid saponins(TSSN)isolated from Dioscorea zingiberensis C.H.Wright(D.zingiberensis)has shown a variety of beneficial bioactivities.However,there are no reports about the neuroprotective effects of the TSSN until now.Therefore,we explored the neuroprotective effects of TSSN on rats against transient focal cerebral ischemia-reperfusion(I/R)and the underlying mechanisms.METHODS The healthy adult Sprague-Dawley rats were randomly assigned into six groups.After pre-treatment with the TSSN intragastrically for six days,the rats were subjected to the ischemia injury by the surgery of middle cerebral artery occlusion(MCAO)for 90 min.Some indexes were evaluated and detected.RESULTS As compared to the I/R group,TSSN group of rats,especially given the 30 mg·kg^-1 of TSSN,not only marked reduction in the neurological deficit scores,cerebral infarct volume,and brain edema,but also an increase in neuron survival(Nissl bodies)in the hippocampal cornuammons 1(CA1)and cortex hemisphere of the ipsilateral ischemia.At the same time,the inflammatory cytokines in serum induced by MCAO were significantly alleviated by the TSSN pre-administration.What′s more,the increase of caspase-3 was evidently reduced in the CA1 and cortex of the hemisphere injured brain.Finally,the down-regulating anti-apoptotic Bcl-2 and up-regulating pro-apoptotic Bax proteins were obviously suppressed.CONCLUSION TSSN plays a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model,and this role may be mediated by its antiinflammatory and anti-apoptotic actions.

Effects of Different Drying Processes on the Effective Components of Stichopus japonicus Viscera

Drying is a basic link in seafood processing, and the effects of forced air drying and vacuum drying on the effective components of Stichopus japonicus viscera were compared with the moisture, total saponin and polysaccharide contents as the detection indexes. The contents of effective components obtained using forced air drying were slightly lower than those obtained using vacuum drying, but the forced air drying method used short drying time and low economic energy consumption. Excellent drying effects and low cost of forced air drying made it can be adapted to the requirements of large-scale production applications.

Comparative pharmacokinetics of five saponins after intravenous administration of TSFS injection and TSFS injection plus TFFG in rats under different physiological states

Sanqi is a popular traditional Chinese medicine and commonly used for promoting blood circulation and removing blood stasis. Notoginsenoside R1, ginsenoside Rg1, Re, Rb1 and Rd are the major active constituents of Sanqi. The purpose of the study was to investigate the pharmacokinetic behavior of the five active constituents from total saponin from Sanqi when it was used in the blood stasis animals or in combination with Gegen. The concentrations of the five active constituents in rat plasma were determined by an ultra-HPLC-ESI-MS/MS method. The main pharmacokinetic parameters were calculated and statistically analyzed using the unpaired student＆#39;s t-test. It was found that the pharmacokinetic parameters of notoginsenoside R1, ginsenoside Rg1 and Rb1 represented a statistically significant difference （Po0.05） between the normal rats and the blood stasis rats after administration of total saponin from Sanqi （TSFS）. And there were statistically significant differences （Po0.05） in the pharmacokinetic parameters of all the five constituents between administration of TSFS alone and combined with total flavonoid from Gegen （TFFG） in blood stasis rats. It suggested that the pharmacokinetic behavior of the active constituents from TSFS could be changed when it was used in blood stasis animals or in combination with TFFG.

Comparative Analysis of Bioactive Components of Polygonatum cyrtonema Hua from Different Areas

Polygonatum cyrtonema Hua from five different areas was used as experimental material,total saponins and total flavonoids.Colorimetric analysis method was used to compare those contents.The results showed that the contents of Polygonatum polysaccharides from different areas were in the range of 8.09%~11.78%,whereas the content of total saponins and total flavonoids was quite different,which were 2.73%~5.01%and 0.21%~0.71%,respectively.In general,these Polygonatum samples had higher polysaccharide content with obvious differences for the contents of total saponins and a low contents of flavones.

Mechanism of TSCS promote anti-Fas antibody-induced FLS apoptosis via blocking MC-tryptase-PAR-2-Rho-FLS signaling pathway

OBJECTIVE It has been supposed that mast cells have important participation in the physiopathology of RA,however,the role of mast cells in the pathogenesis of RA remains unclear.In this study,we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts.METHODS Mast cells and fibroblasts synovial were obtained from mouse.Chemical mediator release was assessed by measuringβ-hexosa-minidase release.TSCS and bone marrow-derived mast cells were co-cultured;the toxic effects of TSCS on mast cell was measured by MTT and CCK-8 method;the releasing amount of tryptase in cell supernatants was measured by Elisa assay;the expression of FLS cell membrane receptor PAR-2 was detected by flow cytometry;the apoptosis of FLS cell was detected through flow cytometry and Western blotting;the level of activated Rho-GTP was detected by the pull-down method and Western blotting.RESULTS In this study,we observed the antiapoptotic effects of tryptase released by mast cell on RA synovial fibroblasts,and found that tryptase significantly increased the expression of PAR-2 on the surface of fibroblast-like synovial cell,significantly activated Rho kinase,significantly inhibited apoptosis of fibroblast-like synovial cell induced by CH11.The release rates ofβ-hexosaminidase and the level of tryptase from bone marrow-derived mast cells after stimulation with different antigen and co-cultured with TSCS significantly decreased compared to the control group.In the co-culture system of mast cells and fibroblast-like synovial cells,TSCS treatment significantly inhibited Rho kinase(P<0.05),significantly promoted apoptosis of fibroblast-like synovial cell induced by CH11(P<0.05).CONCLUSION These results demonstrate that tryptase may play a key role in the physiopathology of RA.TSCS can inhibit mast cells activation and promote FLS cells apoptosis.This provide theoretical and experimental basis for the study of mast cells as targets for new anti-RA drugs.

Effects of Organic and Medium and Trace Element Fertilizers on Yield and Quality of Panax notoginseng

[Objectives]This study aimed to investigate the effects of organic and medium and trace element fertilizers(Ca,Zn,B)on yield and quality of Panax notoginseng to provide theoretical support for rational fertilization in cultivation of P.notoginseng.[Methods]Five fertilization treatments,control(CK),organic fertilizer(OM),zinc fertilizer(ZF),boron fertilizer(BF)and lime(LF),were designed.A two-consecutive-year field plot trail was conducted.The biological traits,yield and saponin content of P.notoginseng were determined.[Results]The application of organic fertilizer had no significant effect on the biological traits of P.notoginseng.Trace element fertilizers significantly increased the scape length of P.notoginseng.Among the treatments,ZF significantly increased the single flower weight but reduced the inflorescence diameter,while the effects of BF were opposite to those of ZF;LF significantly increased the stem thickness and reduced the plant height.All treatments significantly increased the seedling rate of three-year-old P.notoginseng,and the increase in the LF group(20.49%)was the largest,followed by those in the ZF(16.80%)and OM(16.40%)groups,and the increase in the BF group(13.08%)was the smallest.Although OM,ZF and BF treatments caused the root weight of individual plants to decrease,the final yield of each treatment was higher than that of the control group,and the increases in the BF and LF groups exceeded 17%(P<0.05).The total saponin outputs of all the treatments except OM were significantly higher than that of the control group.[Conclusions]Under the conditions of this test,the supplementation of organic and medium and trace element fertilizers on the basis of conventional fertilization will help to increase the yield of P.notoginseng.However,the reduction of the total saponin output of P.notoginseng caused by organic fertilizer cannot be ignored.

Changes in Active Components before and after Microorganism Fermentation of Rhizoma Paridis(Chonglou)

[Objectives] To study the changes in active components before and after microorganism fermentation of Rhizoma Paridis( Chonglou). [Methods]Analytical methods using total saponins and total flavonoids as indicators were established. UV spectrophotometry was used to determine the changes in active components before and after fermentation of Rhizoma Paridis. [Results] The content of total saponins decreased after fermentation,while the content of total flavonoids increased. [Conclusions]The content of total saponins decreased after fermentation and the content of total flavonoids increased. The fermentation process of Monascus purpureus Went. needs further optimization.

Application of Multivariate Analysis Method in Evaluation of Quality and Traits of Sichuan Ophiopogon japonicus

[Objectives] To study the relationship between the yield and total saponin content of Sichuan Ophiopogon japonicus. [Methods]The multi-indicator function model of Sichuan O. japonicus was established by using the multivariate analysis method and taking the yield of and total saponin content of Sichuan O. japonicus as indicators. [Results] In the multivariate non-linear fitting,the Pearson correlation test was used to reduce the dimension of the model,and the significant correlated variables were rejected,leaving three independent variables: the total fresh weight of the plant( x_1),the fresh weight of the aboveground part(x_2) and the fresh weight of fibrous roots(x_3),established the total saponin( y) function model for Sichuan O. japonicus: y = a_1+a_2x_2+ a_3x_3+ a_4x_1x_2+ a_5x_1x_3+ a_6x_2x_3+ a_7x_1~2+ a_8x_2~2+ a_9x_3~2.[Conclusions]When the total fresh weight of plant,fresh weight of aboveground part and fresh weight of fibrous roots were known in Sichuan O. japonicus root tuber,the total saponin content could be estimated by polynomial of these three variables. The establishment of this functional model system is expected to provide a scientific basis for the scientific prediction of the yield and total saponin content of Sichuan O. japonicus.

Total Saponins of Panax notoginseng Activate Akt/mTOR Pathway and Exhibit Neuroprotection in vitro and in vivo against Ischemic Damage

Objective:To reveal the neuroprotective effect and the underlying mechanisms of a mixture of the main components of Panax notoginseng saponins(TSPN)on cerebral ischemia-reperfusion injury and oxygenglucose deprivation/reoxygenation(OGD/R)of cultured cortical neurons.Methods:The neuroprotective effect of TSPN was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)assay,flow cytometry and live/dead cell assays.The morphology of dendrites was detected by immunofluorescence.Middle cerebral artery occlusion(MCAO)was developed in rats as a model of cerebral ischemia-reperfusion.The neuroprotective effect of TSPN was evaluated by neurological scoring,tail suspension test,2,3,5-triphenyltetrazolium chloride(TTC)and Nissl stainings.Western blot analysis,immunohistochemistry and immunofluorescence were used to measure the changes in the Akt/mammalian target of rapamycin(mTOR)signaling pathway.Results:MTT showed that TSPN(50,25 and 12.5μg/m L)protected cortical neurons after OGD/R treatment(P<0.01 or P<0.05).Flow cytometry and live/dead cell assays indicated that 25μg/m L TSPN decreased neuronal apoptosis(P<0.05),and immunofluorescence showed that 25μg/m L TSPN restored the dendritic morphology of damaged neurons(P<0.05).Moreover,12.5μg/m L TSPN downregulated the expression of Beclin-1,Cleaved-caspase 3 and LC3 B-Ⅱ/LC3 B-Ⅰ,and upregulated the levels of phosphorylated(p)-Akt and p-mTOR(P<0.01 or P<0.05).In the MCAO model,50μg/m L TSPN improved defective neurological behavior and reduced infarct volume(P<0.05).Moreover,the expression of Beclin-1 and LC3 B in cerebral ischemic penumbra was downregulated after 50μg/m L TSPN treatment,whereas the p-mTOR level was upregulated(P<0.05 or P<0.01).Conclusions:TSPN promoted neuronal survival and protected dendrite integrity after OGD/R and had a potential therapeutic effect by alleviating neurological deficits and reversing neuronal loss.TSPN promoted p-mTOR and inhibited Beclin-1 to alleviate ischemic damage,which may be the mechanism that underlies the neuroprotective activity of TSPN.

Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

Chemical constituents from acid hydrolyzates of Panax quinquefolius total saponins and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B

Objective:To investigate the hypoglycemic components from the acid hydrolyzates of Panax quinquefolius total saponins,and screen the active compounds by in vitro inhibitory activities toα-glycosidase enzymes and protein tyrosine phosphatase-1 B(PTP1 B).Methods:The hydrolyzates were chromatographed repeatedly over silica gel column,and the structures of the compounds were determined by means of NMR.The in vitro bioassay was performed through the inhibitory effects onα-glucosidase or/and PTP1 B.Results:Eight compounds were isolated,which identified as 20(S)-panaxadiol(1),(20 S,24 R)-dammarane-20,24-epoxy-3β,6α,12β,25-tetraol(2),20(R)-dammarane-3β,12β,20,25-tetraol(3),20(S)-dammarane-3β,6α,12β,20,25-pentol(4),20(R)-dammarane-3β,12β,20,25-tetrahydroxy-3β-O-β-D-glucopyranoside(5),β-sitosterol(6),oleanolic acid(7)and 20(S)-protopanaxadiol(8).Compound 5 was ginseng triterpenoid isolated from the acid hydrolysates of total saponins from P.quinquefolius for the first time.In this paper,the possible in vitro inhibitory activities were investigated.Compound 5 exhibited significantly inhibitory activity againstα-glucosidase,and the IC50 value[(0.22±0.21)μmol/L]was about 43-fold lower than positive control.For the PTP1 B inhibition assay,compound 5 indicated the strongest inhibitory effect with IC50 of(5.91±0.38)μmol/L,followed by compound 4 with IC50 of(6.21±0.21)μmol/L,which were all showed competitive inhibitory pattern by using a Lineweaver-Burk plot.Conclusion:These results supported the potential application of dammaranes from acid hydrolyzates of P.quinquefolius total saponins can be used as ingredients of ancillary anti-diabetic agent or functional factor.