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.

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 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.

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.

Total Saponins of Rubus Parvifolius L.Exhibited Anti-Leukemia Effect in vivo through STAT3 and eIF4E Signaling Pathways

Objective： To investigate the anti-leukemia effect of total saponins of Rubus parvifo/ius L. （TSRP） on K562 cell xenografts in nude mice and the mechanisms of action. Methods： The K562 cell xenografts in nude mice were established, and then randomly divided into 5 groups, the control group, the cytosine arabinoside group（Am-c） and 3 TSRP groups （20, 40 and 100 mg/kg）. The tumor volume and mass of each group of nude mice were measured and the anti-tumor rates of TSRP were calculated subsequently. The apoptosis status of tumor cells was detected by hematoxylin-eosin （HE） and terminal dexynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） staining analysis. Finally, the activities of apoptosis related signaling of signal transducer and activator of transcription 3 （STAT3）, eukaryotic initiation factor 4E （eIF4E） and B-cell lymphoma-2 （bcl-2） were determined with immunohistochemistry tests. Results： Subcutaneous injection of K562 cells induced tumor formation in nude mice, and the TSRP treated group showed a significant inhibitory effect on tumor formation. The nude mice treated with TSRP showed a significant decrease in tumor growth rate and tumor weight in comparison to the control group （all P〈0.05）. The HE staining and TUNEL assay showed that TSRP induced cell death by apoptosis. The immunohistochemical assay showed down-regulation of the bcl-2 gene in the TSRP treated cells. The phosphorylation levels of elF4E and STAT3 were decreased obviously after the treatment of TSRP. Conclusion： TSRP had an excellent tumor-suppressing effect on K562 cells in the nude mice xenograft model, suggesting that TSPR can be developed as a promising anti-chronic myeloide leukemia drug.

A mathematical model for quality evaluation of total saponins of Panax japonicas based on hypolipidemic activity

Objective: The chemical finger printing-based methods for evaluating TCMs quality can report partial of TCMs quality without linking to effective constituents. In this study, a mathematical model was established for the quality evaluation of total saponins of Panax japonicus(TSPJ), a folk medicine in China and Japan for treating diseases, through coupling the dynamic changes of chemical constitutions with corresponding activities.Methods: High-performance liquid chromatography(HPLC) fingerprints were applied to establish the chromatographic database of TSPJ. The associated hypolipidemic activity database was determined by TG assay using Hep G2 cell model. Correlation analyses of two databases were performed by partial least squares(PLS) for calculating regression coefficients, and the interval value of YZL value(the ratio of positive and negative peak-to-peak area coefficient) closely related to hypolipidemic activity was refined by the formula of Norminv function to value the quality of TSPJ.Results: In this study, the chromatographic data of 16 common peaks were obtained from 20 batches of TSPJ. After the estimate by this mathematical evaluation model, seven peaks were positively correlated with hypolipidemic activity, and nine peaks were negatively correlated with hypolipidemic activity.When the YZL value was less than 0.7861, the quality of sample was inferior, while YZL value was more than 6.6992, and the quality of samples was superior. The quality of another ten batches of TSPJ was further assessed to verify this method.Conclusion: These results indicated that the established model could be usefully applied to evaluate the quality of TSPJ in the hypolipidemic activity.

The Effects of Total Saponins of Panax Ginseng on Hematopoietic Progenitor Cells in Healthy Humans and Aplastic Anemia Patients

Ginseng is said to have beneficial effects on anemia. The proliferation effects of totalsaponins of Panax ginseng (TSPG) on hematopoietic progenitor cell in healthy individuals and 29 patientswith aplastic anemia (AA) were observed through bone marrow cultures of burst forming unit-erythroid(BFU-E) , colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) in vitrcacompared with methyltestosterone (MT). The results suggest TSPG might prompt the prolif-eration of normal progenitor cellS at a concentration of 20 g/ml. The numbers of BFU-E ,CFU-E and CFU-GM increased by 37. 8±2.9 % , 31. 4±2. 9 % and 33. 3± 4. 0 % respectively over the controls ; further-more TSPG was still useful to BFU-E,CFU-E growth without Epo in vitro, although the colony nurnberswere much lower. Otherwise MT was useless to CFUGM. Of the 29 patients with AA, 14 who respondedto MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30 % ) ,but immune-mediated AA (patient's peripheral blood mononucleated cell suppressed normalhematopoiesis) and stem cell decreased AA (few of colonies were formed) showed almost no expressionfor TSPG activity because of the immunological suppression system and the absence of progenitors.

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.

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.

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.

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.

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.

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.

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.

Attenuation of Brain Inflammatory Response after Focal Cerebral Ischemia/Reperfusion with Xuesaitong Injection (血塞通注射液) in Rats

Objective： To investigate the neuro-protective effect of Xuesaitong Injection （血塞通注射液, XST） on brain inflammatory response after transient focal cerebral ischemia/reperfusion in rats. Methods： Focal cerebral ischemia/reperfusion models of male rats were induced by transient occlusion for 2 h of middle cerebral artery （MCA） which was followed by 24 h reperfusion. XST was administered through intraperitoneal injection of 25 mg/kg or 50 mg/kg at 4 h after the onset of ischemia. After reperfusion for 24 h, the neurological function score was evaluated, the brain edema was detected with dry-wet weight method, the myeloperoxidase （MPO） activity and the expression of intercellular adhesion molecule-1 （IOAM-l） of ischemic cerebral cortex and caudate putamen was determined by spectrophotometry and immunohistochemistry respectively. Results： XST not only lowered neurological function score at the dose of 50 mg/kg, but reduced brain edema and inhibited MPO activity and IOAM-1 expression as compared with the ischemia/reperfusion model group （P〈0.01）. Conclusion： XST has a definite effect on inhibiting the expression of IOAM-1 and neutrophil infiltration in rats with cerebral ischemia/reperfusion when treatment started at 4 h after ischemia onset, and also attenuates inflammation in the infarcted cerebral area.

Rubus Parvifolius L.Inhibited the Growth of Leukemia K562 Cells in Vitro and in Vivo

Objective：To determine the antiproliferative activity of Rubus parvifolius L.（RP）extract,its medicinal serum and RP total saponins（RPTS）against K562 cells in vitro and in vivo.Methods：Nude mice models bearing leukemia tumors were treated with different concentrations of RP extract.The size,weight and histopathological change of leukemic tumors were determined.Semi-solid agar culture and methylthiazolyl tetrazolium（MTT）assay were used to determine in vitro the inhibition of colony formation and proliferation of K562 cells respectively by different concentrations of RP medicinal serum and RPTS.Results：RP extract had a tumor inhibition rate of 84.8%when administered to mice at a dose of 1.0 g/day of crude RP root equivalent.Semi-solid agar culture of K562cells in the presence of 20%（v/v）of RP medicinal serum and 150 mg/L RPTS demonstrated a 50.8%and 100%inhibition of the colony forming unit（CFU）-K562,respectively.The same doses of RP medicinal serum and RPTS showed a proliferation inhibition of 31.4%and 86.3%,respectively against K562 cells in MTT assay.Conclusion：RP extract and RPTS show effective antiproliferative activity against myeloid leukemia cells in vitro and in vivo.