Preliminary study on response and its mechanism of ginsenoside biosynthesis in Panax ginseng to water regulation / 中国中药杂志

The study is aimed to explore the effect of soil moisture content on ginsenoside biosynthesis and explain its mechanism from the perspectives of antioxidant enzyme system and gene expression of key enzymes in the pathway of ginsenoside synthesis. In the study,two years old Panax ginseng was used as the experimental material and three moisture gradient,40% of saturated water content( W1),60%( W2),80%( W3) were set up. The content of 11 monomeric saponins were determined by HPLC. With GAPDH as a reference gene,six key enzymes( HMGR,SS,β-AS,CYP716 A47,CYP716 A52 v2,CYP716 A53 v2) in ginseng saponin synthesis pathway expression were analyzed by fluorescent quantitative PCR and the activities of superoxide dismutase( SOD),peroxidase( POD),catalase( CAT) activity and MDA content were also determined. With the increase of soil water,the content of ginseng saponin and biomass showed an increasing trend. PPD( Rb1,Rc,Rb2,Rd,Rh2,Rb3,Rg3),PPT( Rg1,Re,Rf) ginsenoside,Ro and total ginsenoside reached the maximum value on August 30,were 9.92,5.48,0.63 mg·g-1,respectively. During the whole regulation period,the antioxidant activity of W3 was greater than that of W1,and the MDA content was less than that of W1. At W3,expression levels of β-AS,CYP716 A47 and CYP716 A53 v2 showed an increasing trend,while HMGR and SS genes showed relatively stable expression levels under various water conditions. According to the correlation analysis,HMGR and SS genes in the W3 treatment group were significantly positively correlated with PPD,PPT ginsenoside and Ro,CYP716 A52 v2 gene was significantly positively correlated with Ro,and CYP716 A47 gene was significantly positively correlated with PPD ginsenoside. There was a significant positive correlation between β-AS gene and PPD ginsenoside in W1 and W2 treatment. Therefore,W3 is the optimum moisture content,ginseng total saponins and monomer saponin content is the highest,the gene closely correlation with content of saponins and more conducive to the accumulation of ginsenosides.

Research on Chinese medicine pairs effects of Panax notoginseng and Bletilla striata before and after compatibility on contents of notoginsenosides / 中国中药杂志

To discuss the synergistic mechanism of compatible use of two medicinal herbs,Panax notoginseng and Bletilla striata, an HPLC was established to determine two ginseng saponins (20S)-ginseng saponin Rg₃ and ginseng saponin Rh₄ contained in single decoction of Panax notoginseng as well as in compound decoction of Panax notoginseng and Bletillastriata in different compatibility ratio (1∶0.5, 1∶1, 1∶2), followed by analyzing the impact of amount of notoginsenosides after compatibility. As a result, compared with the single decoction of Panax notoginseng, the contents of ginseng saponin Rg₃ and ginseng saponin Rh₄ in the compound decoction of Panax notoginseng and Bletillastriata were on the rise as the increasement of the amount of Bletillastriata. The contents of the notoginsengsaponin R₁, ginseng saponin Rg₁ and ginseng saponin Rb₁ of Panax notoginseng single decoction were significantly decreased after compatibility. Therefore, after compatibility, it was more easy to produce (20S)-ginseng saponin Rg₃ and ginseng saponin Rh₄.This study can extend to a method of preparation of (20S)-ginseng saponin Rg₃ and ginseng saponin Rh₄. Furthermore, after compatibility, two ginseng saponins which had lipase inhibitory effect were both increased significantly, indicating that the compatibility of these two herb medicines may have effect on losing weight.

The Effects of Ginseng Saponin on Relaxation of Smooth Muscle in the Lower Urinary Tract and the Corpus Cavernosum / 영남의대학술지

BACKGROUND: Korean ginseng (KG) has been used as a general tonic, and for voiding dysfunction for a long time in oriental society. However, scientific basic studies on the use of KG, have been rare, especially for voiding and erectile dysfunction. This study was performed to investigate the effects of KG on voiding and erectile function by examining the effects of total saponin (TS) on the bladder, urethral and penile cavernosal smooth muscle. MATERILAS AND METHODS: To examine the effects of TS, NewZeland white rabbits were used to obtain tissue strips from the smooth muscle of the bladder, proximal urethra and corpus cavernosum. Adult Sprague Dawley rats were used to examine the changes in urodynamic findings and penile erection after administration of TS. RESULTS: In proximal urethral strips, the rate of relaxation of the proximal urethra was increased from 9.0+/-2.9 to 33.7+/-4.8% in a dose-dependent manner when the concentration of TS was added accumulatively from 0.25 mg/ml to 4.0 mg/ml (p<0.05). However, no significant response was observed in the bladder strips within these concentration ranges. For the corpus cavernosal strips, the rate of relaxation ranged from 5.8+/-2.1 to 36.7+/-5.8%, increasing in a dose-dependent manner when TS was increased from 1.0 mg/ml to 4.0 mg/ml (p<0.05). After administration of 0.1 ml of TS (32 mg/ml) in the rat, the bladder pressure was 37.5+/-8.5 mmHg at 52.1+/-7.0 sec. during isovolumetric bladder contraction, showing no significant differences from 35.7+/-7.8 mmHg and 50.7+/-7.2 sec, respectively, before treatment. However, when 0.1 ml of TS (32 mg/ml) was administered, the relative reduction of urethral pressure was 6.9+/-0.5 mmHg at 62+/-7.5 sec, which was significantly higher compared to 4.6+/-1.1 mmHg at 45+/-10 sec before treatment (p<0.05). For the cavernosal injection study, the change in intracavernosal pressure (delta ICP) was examined after administering 0.1 ml of TS. The cumulative additions of TS at concentrations from 0.5 mg/ml to 32 mg/ml increased delta ICP from 1.3+/-0.5 to 21.3+/-7.8 mmHg in a dose-dependent manner (p<0.05). The duration of tumescence was from 0.3+/-0.1 to 5.2+/-0.2 min, showing dose-dependent increase (p<0.05). Furthermore, the cumulative addition of TS at concentrations from 0.5 mg/ml upto 32 mg/ml did not cause any significant change in systemic blood pressure. CONCLUSION: These results suggest that ginseng improves voiding functions, which is mainly achieved by TS relaxing the proximal urethra, the most important part of the bladder outlet function. In addition, ginseng safely induced a penile erection hemodynamically by relaxing the corpus cavernosum.

Inhibition of MMP-13 mRNA expression by ginseng saponin in fetal rat calvarial cells / 대한치주과학회지

There is a potential role of collagenase-3 in alveolar bone loss and periodontal disease progression, we need to develope or find chemotherapeutic drugs or herbal agents which may regulate the expression of MMP-13. Ginseng saponin, one of the major components of Korea ginseng(panax ginseng) root, has many various biologic effects, such as cytotoxic effect, tumoricidal effects, cytokine regulations, and protein biosynthesis effect. The purpose of this study was to determine the effects of Korea red ginseng saponin on MMP-13 gene expression in osteoblasts. The experimental groups were cultured with ginseng saponin in concentration of 1.0, 10, 25, 50, 100, 250 and 500microgram/ml for MTT assay. Primary rat calvarial cells were pre-treated for 1 hour with ginseng saponin(100 microgram/ml) and then stimulated with IL-1beta(1.0ng/ml) and PTH (10 nM). MMP-13 gene expression was evaluated by RT-PCR. The results were as follows: Ginseng saponin was cytotoxic to osteoblast at concentration exceeding 250microgram/ml for longer than 24 hours in tissue culture(p<0.01). In RT-PCR analysis, steady state MMP-13 mRNA levels were increased approximately 350% by IL-1beta, and 400% by PTH when normalized to untreated control. IL-1beta-indued MMP-13 mRNA expression was reduced 50% by pre- treatment with ginseng saponin. But ginseng saponin didn't inhibit MMP-13 expression from PTH stimulated cells. This results suggest that ginseng saponin inhibit IL-1beta-indued MMP-13 mRNA expres- sion.

Effects of Ginseng Saponin on the Stress-Induced Plasma Corticosterone Levels in Mice / 신경정신의학

OBJECTIVES: The present study was designed to investigate the effect of ginseng saponin and its major active metabolite on the HPA axis under acute stress-i.c.v. injection stress, and immobilization stress, and to examine whether nitric oxide is involved in the mechanism of ginseng saponin on the HPA axis under acute stress. METHODS: In the experiment to study the effect of ginseng on HPA axis during stress, various dose of GTS were injected intracerebroventricularly(i.c.v.) or intraperitoneally(i.p.). Plasma corticosterone levels were measured 30 min after the i.c.v. injection stress. Immobilization stress was applied for 30 min and then blood was cellected for the assays of plasma corticosterone levels immediately after the completion of immobilization stress. To determine the active ginsenosides that can affect the stressinduced plasma corticosterone levels, various dose of each gisendosides(Rb1, Rb2, Rc, Re, Rf, Rg1, 20(S)-Rg3, and 20(R)-Rg3) were injected i.c.v. or i.p.. In the experiment to determine the involvement of the nitric oxide in the inhibitory effect of ginseng on the HPA, NG-Nitro-L-arginine methyl ester(L-NAME) and ginsenosides were coadministered i.c.v. or i.p., and plasma corticosterone levels were measured 30 min after stress was applied. RESULTS: First, the present study showed that ginseng total saponin, ginsenoside Rg3(S form), and ginsenoside Rc administered i.c.v. attenuated the intracerebroventricular injection stress-induced increase in plasma corticosterone levels, and these effects were removed by nitric oxide co-injection. Second, ginseng total saponin and ginsenoside Rc administered i.p. attenuated the immobilization stress-induced increase in plasma corticosterone levels, but ginsenoside Rg3(S form) did not attenuate the immobilization stress-induced increase in plasma corticosterone levels. The attenuative effects of ginseng total saponin and ginsenoside Rc in the immobilization stress-induced increase in plasma corticosterone levels were not affected by L-NAME co-injection. CONCLUSION: This study suggests that ginseng saponin attenuated stress-induced increase in plasma corticosterone levels and these effects were mediated by different mechanisms according to the components of ginseng saponin, and routes of administration.

The Harmful Effects of Superoxide Radical and the Protective Effect of Ginseng Saponin on Rat Whole Bladder Contractility / 대한비뇨기과학회지

PURPOSE: We investigated the effects of the superoxide radical on rat whole bladder contractility with duroquinone (superoxide radical generator, Dq) and diethyldithiocarbamate (superoxide dismutase inhibitor, DETCA), and the effects of ginseng saponin (GS) against superoxide radical injury. MATERIALS AND METHODS: Isometric tension changes of isolated rat whole bladders were recorded in an organ bath using a force transducer. The acute effects of Dq and Dq preincubated with DETCA were assessed on resting tension, electrical field stimulation, and bethanechol-, ATP-, and KCl-induced contraction. The effects of Dq and Dq preincubated with DETCA in the presence of sodium nitroprusside and GS were investigated. RESULTS: The resting tension of the muscle was not changed by Dq and Dq preincubated with DETCA. Dq had a harmful effect on only ATP- and KCl-induced detrusor contraction, whereas Dq pretreated with DETCA attenuated the induction of detrusor contraction which was reduced in response to the exogenous NO including GS. In the presence of L-NAME, the effects of GS reduced the Dq-induced inhibition on the detrusor contractility. CONCLUSIONS: It is suggested that the superoxide radical may be the cause of voiding difficulty. GS, as a NO synthesis stimulator, seems to act as a scavenger of the superoxide anion. However further study on the effect of each subfraction of GS is needed for clinical application.

Determination of Ginseng Saponin Rg_1,Rb_1,Re and Notoginseng Saponin R_1 in Tangzhiqing Capsule by HPLC / 中药新药与临床药理

Objective To establish a method of determining effective components in Tangzhiqing Capsule.Methods Gin- seng saponin Rg_1,Rb_1,Re and notoginseng saponin R_1 in the capsule were separated and purified by D_(101)macroporous absorption resin,and then determined by HPLC.Results The linearity arrange of ginseng saponin Rg_1,Re,Rb_1 and no- toginseng saponin R_1 were 1.88～11.28?g,1.76～10.56?g,0.294～1.764?g,0.752～2.256?g and the recov- eries were 101.51%(RSD=0.75%),100.58%(RSD=0.46%),100.29%(RSD=1.01%),98.64% (RSD = 0.73%)respectively.Conclusion The method is simple,feasible and reproducible,and can be used for the determination of effective components in Tangzhiqing Capsule.

Antioxidative Effect of Ginseng Saponin on Cardiac Endothelial Cells in Culture / 대한해부학회지

The cytotoxic effects of oxygen free radicals and the antioxidative effect of ginseng saponin (SPN) on cardiac endothelial cell cultures derived from 3-day old rats were studied. Reactive oxygen species were generated by hypoxanthine (HX) and xanthine oxidase (XO) mixture to the culture medium. Exposure of cardiac endothelial cells to this oxygen-radical-generating system resulted in significant time-dependent decrease of MTT activity and increase of lactate dehydrogenase (LDH) release. These results correlated well with the morphological examination of randomly selected cultured cardiac endothelial cells, which showed large cytoplasmic vacuoles, disordered organelles, pronounced increase of endoplasmic reticular swelling, and decreased maintenance of membrane integrity. The decrease in cell viability and increase of LDH release induced by the oxygen free radicals in cardiac endothelial cell cultures were blocked during the first two hours by antioxidants such as ginseng saponin (SPN), deferoxamine (DFX), and ginseng saponin/deferoxamine mixture (SPN/DFX). These antioxidative effects were significantly greater in the SPN-treated group than in the other antioxidant-treated groups. Especially, the cells of the SPN-treated group showed well developed cytoskeletons, which enabled them to firmly attach to the culture vessel. In conclusion, these results indicate that ginseng saponin has a significant antioxidative effect on cardiac endothelial cells in culture and plays an important role in stimulating the formation of cytoskeleton and maintaining the integrity of cell membrane.

Influence of Total Ginseng Saponin on Contractile Responses of Vasoconstrictors in the Isolated Rat Aorta

BACKGROUND: It has been known that Ginseng extract causes the hypotensive action while it rather produces the hypertensive action. Some studies have suggested that Ginseng extract causes a biphasic response on blood pressure, namely, transient fall followed by prolonged elevation. It has been also shown that administration of Korean Red Ginseng powder has no effect on blood pressure in normotensive and hypertensive rats. The present study was designed to examine the effect of total Ginseng saponin on contractile responses of vasoconstrictors in the rat aorta and to establish the mechanism of its action. METHODS: The ring segment of aorta was mounted in a muscle bath filled with oxygenated Krebs solution for the measurement of isometric tension. After the equilibration period, under the presence of total Ginseng saponin, isometric tension induced by some vasoconstrictors were observed and compared to the control responses. The data were expressed as % of the control tension. RESULTS: Phenylephrine (an adrenergic alpha1-receptor agonist) and high potassium (a membrane depolarizing agent) caused greatly contractile responses in the rat aorta, respectively. However, in the presence of total ginseng saponin (600 g/ml), the contractile responses of phenylephrine (10(-6) and 10(-5) M) and high potassium (3.5 x 10(-2) and 5.6 x 10(-2) M) were markedly potentiated whereas prostglandin F2alpha(5 x 10(-6) M)-induced contractile responses was not affected. The contractile responses induced by phenylephrine (10(-5) M) and high potassium (3.5 x 10(-2) M) even under the presence of total ginseng saponin (600 g/ml) were greatly inhibited by the pretreatment of nicardipine (10(-6) M), a calcium channel blocker. CONCLUSION: Taken together, these experimental results suggest that total ginseng saponin can enhance the contractile responses evoked by stimulation of adrenergic alpha1-receptor and the membrane depolarization in the isolated rat aortic strips, which seems to be associated to calcium influx.

The Effects of Ginseng Saponin on Rabbit Cavernosal Smooth Muscle Relaxation in vitro / 대한비뇨기과학회지

PURPOSE: Ginseng saponin, the active ingredients extracted from Panax ginseng, has been used as aphrodisiacs in oriental country, however the exact action mechanism of ginseng saponin still remains uncertain. This study was undertaken to see if ginseng crude saponin(GCS) induce the L-arginine/Nitric oxide(NO) pathway related the corpus cavernosal smooth muscle relaxation, and to obtain the fundamental data for clinical application. MATERIALS AND METHODS: Isometric tension changes of rabbit corpus cavernosal strip to various drugs and electrical stimulation in an organ chamber(volume:30m1) were recorded with a pressure transducer. RESULTS: GCS(0.2-8.0mg) relaxed the smooth muscle of rabbit corpus cavernosum (SMRCC) precontracted with phenylephrine(10(-5)M) in dose-dependent manner GCS at the concentration of 0.75mg significantly enhanced the relaxation of corpus cavernosum induced by the electrical field stimulation. GCS(0.2-8.0mg)-induced relaxation was significantly attenuated by atropine(10(-6)M), methylene blue(10(-4)M) and L-NAME(10(-5)M), but there was no significant change of GCS-induced relaxation of SMRCC by adding VIP antagonist or indomethacin. In addition, the inhibition of GCS-induced relaxation of SMRCC by methylene blue or L-NAME was greater than that by atropine. CONCLUSIONS: These data suggest that GCS, as a NO donor, induces the relaxation of SMRCC through the L-arginine/NO pathway. For the clinical application of GCS, further studies should be needed to clarify the subfraction of GCS.

The Effect of Ginseng Saponin on the Dopaminergic Neurons in the Parkinson's Disease Model in Mice

Saponin has been known to be a major antioxidant component in panax ginseng. Recent experimental study suggests that some antioxidant materials prevent Parkinson's disease caused by 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP) in an animal model. The present study was performed to demonstrate the effect of ginseng saponins in the Parkinson's disease model induced by MPTP. To verify the effect of ginseng saponin on dopaminergic neurons in the mice brain, the tyrosine hydroxylase-immunoreactive (TH-ir) neurons were observed by immunohistochemical stain and immunoelectron microscopy (preembedding method). Also, in order to estimate the immunoreactivity of dopaminergic neuropils, they were quantified by image analysis. The number of TH-ir neurons of substantia nigra was significantly increased in the high-dose (0.46 mg/kg) ginseng saponin group compared with the MPTP injected group. The immunoreactivity of TH-ir neuropils in striatum was significantly increased in both high and low-dose (0.1 mg/kg) ginseng saponin groups compared with the MPTP injected group. In immunoelectron microscopic observation, TH-ir neurons of the control and both ginseng saponin injected group showed normal nuclei and well preserved cytoplasmic organelles. In the MPTP injected group, dying dopaminergic neurons showed destroyed nuclei and cytoplasmic organelles. These results suggest that ginseng saponin has a protective effect on the Parkinson's disease model induced by MPTP.

Determination of ginseng saponin Rg_1,Rb_1 and notoginseng saponin R_1 in Qianjin Shenanning Pills by HPLC / 中成药

AIM:To establish a method of determining effective components in Qianjin Shenanning Pills(Flemingia Philippinensis Merr.et Rolfe,Radix et Rhizoma Ginseng rubra,Radix et Rhizowa Notoginseng,etc). METHODS: The contents of ginseng saponin Rg_1,Rb_1 and notoginseng saponin R_1 were determined by HPLC. RESULTS: The four effective components could be separated and purified by macroporous adsorptive resins of D_(101) and n-butanol,and their contents could be determined. CONCLUSION: The method is simple,feasible and reproducibility is good,it can be used as quality control in medicinal preparation.

INFLUENCE OF GINSENG SAPONIN ON THE CIRCADIAN RHYTHM IN BRAIN CATECHOLAMINE / 中国药理学通报

The circadian levels of norepinephrine ( NE ) and dopamiae (DA) in the whole brain were determined spectrofluorometrically in control mice and in mice pretreated with ginseng saponin. All animals were adapted for a minimum period of 2 weeks to an environmental room equipped with, an automatically timed 12h-light and 12h-dark illumination cycle before experiment. Data obtained at 4 h intervals for 48h spans were analyzed by the mean cosinor method.It has been proved that the catecholamine concentrations in the brain vary diurnally. The whole brain NE and DA contents were highest during the middle of dark phase and decreased to a low levels after the onset of light phase. In administration of ginseng saponin altered the circadian pattern and ( or ) levels of brain catecholamine over controls. In order to check the above results reserpine was injected to mice. The levels of NE and DA remarkedly depleted at the times studied, and they showed a significant circadian rhythms as well.It seems that ginseng saponin selectively modulates the circadian variations of brain NE and DA, and its effects may be as a cosinor function of the time of day.