Repeated-dose 26-week oral toxicity study of ginsenoside compound K in Beagle dogs.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginsenoside compound K (CK), a product produced by the intestinal bacteria-mediated breakdown of ginsenoside, exhibits a wide array of pharmacological activities against diverse targets. However, few of preclinical safety evaluation of CK is reported. AIMS OF THE STUDY: The present study therefore sought to assess the toxicity of oral CK in Beagle dogs over a 26-week period. MATERIAL AND METHODS: All dogs received 4, 12, or 36 mg/kg oral CK doses for 26 weeks with regular monitoring, followed by a 4-week recovery period. Animals were monitored through measurements of temperature, weight, food intake, blood chemistry and hematological findings, electrocardiogram (ECG) measurements, urinalysis, gross necropsy and organ weight and tissue histopathology. RESULTS: Animals in the 36 mg/kg group exhibited an apparent reduction in body weight over the study period, in addition to the presence of focal liver necrosis and increased plasma enzyme levels (alanine aminotransferase, ALT; alkaline phosphatase, ALP) consistent with hepatotoxicity, although there was some evidence suggesting this toxicity was reversible. Animals in the 4 and 12 mg/kg groups did not exhibit any apparent toxicity for any measured parameters. CONCLUSION: These results thus indicate that the no observed adverse effect level (NOAEL) in dogs is 12 mg/kg.

Three New Triterpenoid Saponins from Notoginseng Medicinal Fungal Substance.

Three new triterpenoid saponins, named ginsenoside-Rh23 (1), ginsenoside-Rh24 (2), and ginsenoside-Rh25 (3), were isolated from notoginseng medicinal fungal substance. Their structures were elucidated by a combination of 1D- and 2D-NMR, MS and chemical analysis. Compounds 1 - 3 exhibited moderate cytotoxic activity against MCF-7 and NCI-H460 cancer cell lines.

Panax ginseng and Panax quinquefolius: From pharmacology to toxicology.

The use of Panax ginseng and Panax quinquefolius in traditional Chinese medicine dates back to about 5000 years ago thanks to its several beneficial and healing properties. Over the past few years, extensive preclinical and clinical evidence in the scientific literature worldwide has supported the beneficial effects of P. ginseng and P. quinquefolius in significant central nervous system, metabolic, infectious and neoplastic diseases. There has been growing research on ginseng because of its favorable pharmacokinetics, including the intestinal biotransformation which is responsible for the processing of ginsenosides - contained in the roots or extracts of ginseng - into metabolites with high pharmacological activity and how such principles act on numerous cell targets. The aim of this review is to provide a simple and extensive overview of the pharmacokinetics and pharmacodynamics of P. ginseng and P. quinquefolius, focusing on the clinical evidence which has shown particular effectiveness in specific diseases, such as dementia, diabetes mellitus, respiratory infections, and cancer. Furthermore, the review will also provide data on toxicological factors to support the favorable safety profile of these medicinal plants.

Impairment of preimplantation and postimplantation embryonic development through intrinsic apoptotic processes by ginsenoside Rg1 in vitro and in vivo.

Ginsenoside Rg1, which is the most abundant compound found in Asian ginseng (Panax ginseng), has demonstrated various pharmacological actions, including neuroprotective, immune-stimulatory, and antidiabetic effects. Pregnant women, especially in the Asian community, consume ginseng as a nutritive supplement. Thus, the effects of ginsenoside-Rg1 on embryonic development need to be investigated, such as in a mouse model. As previous investigations have found that ginsenoside Rg1 appears to either trigger or prevent apoptosis in different cell lines, the effects of this agent on apoptosis remain to be clarified. In this study, we investigated whether ginsenoside Rg1 exerts a hazardous effect on mouse blastocysts and/or affects subsequent embryonic development in vitro and in vivo. Blastocysts treated with 25-100 µM ginsenoside Rg1 exhibited significant induction of apoptosis and a corresponding decrease in the inner cell mass (ICM) cell number. Importantly, the implantation rate was lower among ginsenoside Rg1-treated blastocysts compared to untreated controls. Moreover, embryo transfer assays revealed that blastocysts treated with 100 µM ginsenoside Rg1 exhibited increased resorption of postimplantation embryos and decreased weight among surviving fetuses. In vivo, intravenous injection of mice with ginsenoside Rg1 (2, 4, or 6 mg/kg body weight/day) for 4 days was associated with increased apoptosis of blastocyst-stage embryos and negatively impacted early embryonic development. Further experiments revealed that these effects may reflect the ability of ginsenoside Rg1 to trigger oxidative stress-mediated intrinsic apoptotic signaling. Our in vitro results indicate that ginsenoside　Rg1 treatment increases intracellular oxidative stress, decreases mitochondrial membrane potential, increases the Bax/Bcl-2 ratio, and activates caspase-9 and caspase-3, but not caspase-8. Taken together, our study results strongly suggest that ginsenoside Rg1 induces apoptosis and impairs the early preimplantation and postimplantation development of mouse embryos, both in vitro and in vivo.

Potential accumulation of protopanaxadiol-type ginsenosides in six-months toxicokinetic study of SHENMAI injection in dogs.

SHENMAI injection (SMI), derived from famous Shen Mai San, is a herbal injection widely used in China. Ginsenosides are the major components of SMI. To monitor the exposure level of SMI during long-term treatment, a 6-month toxicokinetic experiment was performed. Twenty-four beagle dogs were dived into four groups (n = 6 in each group): a control group (0.9% NaCl solution) and three SMI groups (2, 6 or 3 mg/kg). The dogs were i.v. infused with vehicle or SMI daily for 180 d. Blood samples for analysis were collected at specific time points as follows: pre-dose (0 h); at 10, 30, and 60 min during infusion; and at 10, 30, 60, 90, 120, 240, and 300 min post-administration. Concentrations of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1 in the plasma were determined simultaneously by liquid chromatography-tandem mass spectrometry. Non-compartmental parameters were further calculated and analyzed. Significant differences were found between the kinetic behavior of 20(S)-protopanaxadiol-type (PPD-type) and 20(S)-protopanaxatriol-type (PPT-type) ginsenosides. Increasing in the exposure level of PPD-type ginsenosides was observed in dogs during the experiment. Therefore, PPD-type ginsenosides are closely related to the immunity modulation effect of SMI. Increased PPD-type ginsenoside exposure level may present potential toxicity and induce drug-drug interaction risks during SMI administration. As such, PPD-type ginsenoside accumulation must be carefully monitored in future SMI research.

The Safety Evaluation of Salvianolic Acid B and Ginsenoside Rg1 Combination on Mice.

Our previous study indicated that the combination of salvianolic acid B (SalB) and ginsenoside Rg1 (Rg1), the main components of Salvia miltiorrhizae and Panax notoginseng, improves myocardium structure and ventricular function in rats with ischemia/reperfusion injury. The present study aimed to determine the safety of the combined SalB and Rg1 (SalB-Rg1) in mice. The safety of SalB-Rg1 was evaluated through acute toxicity and repeated-dose toxicity. In the acute toxicity study, the up and down procedure was carried out firstly, and then, the Bliss method was applied. In the toxicity study for seven-day repeated treatment of SalB-Rg1, forty Kunming mice were randomly divided into four groups. The intravenous median lethal dose (LD50) of the SalB-Rg1 combination was 1747 mg/kg using the Bliss method. For both the acute toxicity study and the seven-day repeated toxicity study, SalB-Rg1 did not induce significant abnormality on brain, heart, kidney, liver and lung structure at any dose based on H&E stain. There were no significant changes related to the SalB-Rg1 toxicity detected on biochemical parameters for two kinds of toxicity studies. The LD50 in mice was 1747 mg/kg, which was more than one hundred times higher than the effective dose. Both studies of acute toxicity and seven-day repeated dose toxicity indicated the safety of the SalB-Rg1 combination.

[Autotoxic effect of ginsenoside extrats on growth of American ginseng in different medium].

Ginsenosides are the abundant secondary metabolites in American ginseng (Panax quinquefolium), it could be released into soil through root exudation and decomposition during plant growth. This study determined ginsenoside contents in American ginseng cultivated soil by HPLC. Three ginsenosides, Rb1, Rb2 and Rd, were detected in the rhizosphere soil of 3-4 years old American ginseng cultivated in Huairou District, Beijing, and their contents were 0.80-3.19 mg x kg(-1). Correspondingly, the contents of the three ginsenosides in soil solution were 4-16 mg x L(-1) at field water-holding capacity of 20%. According to the field soil test data, we designed the concentration of ginsenosides for bioassays (0.2-125 mg x L(-1) in solution or 0.2-125 mg x kg(-1) in soil). The results showed that radicle lengths of American ginseng were reduced by 6%-23% in solution containing 0.2-125 mg x L(-1) ginsenoside extract, and a significant difference was observed at concentration of 125 mg x L(-1) (P < 0.05). The shoot lengths of American ginseng were not significantly inhibited by 0.2-125 mg x L(-1) ginsenosides extractions. After 20 days of growth in nutrient solution amended with 25 mg x L(-1) ginsenosides extraction, plant height of 3-year-old American ginseng seedling was decreased by 28% compared to the control, and the biomass of aerial parts was also reduced by 50% (P < 0.05). However, the growth of newly-grown fibrous root was not significantly inhibited. Comparatively, when American ginseng embryos were cultivated into sterile or non-sterile soil, neither radicle lengths nor shoot lengths were significantly affected by 0.2-125 mg x kg(-1) ginsenoside extracts. In conclusion, ginsenosides showed autotoxic effect on growth of American ginseng radicle and adult seedling, however, this effect was weakened in field soil.

One-month toxicokinetic study of SHENMAI injection in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: 'SHENMAI' injection (SMI) has been widely used in cardioprotection and modulation of the immune system because of its great efficacy. SMI primarily comprises the saponins from Panax ginseng and Ophiopogon japonicas. The profiles of saponins in SMI during long-term toxicokinetics remain unclear. MiR-146a possesses excellent sensitivity as a bio-marker in the innate immunity modification effect of SMI. AIM OF THE STUDY: Is to monitor the exposure level of SMI during a one-month toxicokinetic experiment, an analytical method involving ESI-LC-MS/MS technology was developed to determine 20 (S)-protopanaxadiol-type ginsenoside (Rb1, Rb2, Rc, Rd), 20 (S)-protopanaxatriol-type ginsenoside (Rg1, Re, Rf), oleanolic acid-type ginsenoside (Ro), and ophiopogonin D in rats. The levels of AST, CK, ALT, SOD, GSH-pX, MDA, miR-146a, and ECG were measured to explore the effects of SMI in cardiologic function and immune activity. RESULTS: Results show that the levels of AST, CK, and MDA decreased upon the administration of SMI. The level of miR-146a increased upon the administration of SMI dosage. During the administration of SMI, increasing exposure levels of 20 (S)-protopanaxadiol-type ginsenosides were also observed. CONCLUSION: The 20 (S)-protopanaxadiol-type ginsenosides were considered potential PK/TK markers because of their high exposure levels that continuously increased. Oxidative stress was slightly alleviated during the toxicokinetic study. Based on the level of miR-146a, negatively regulated innate immunity was observed. The regulation became more serious with increasing exposure levels of 20 (S)-protopanaxadiol-type ginsenosides. Negatively regulated innate immunity could be induced by long-term administration of SMI (>0.4g/kg).

Chronic toxicity of ginsenoside Re on Sprague-Dawley rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginseng has been widely used for hundreds of years in both China and other countries. It is well accepted that the pharmacological effects of ginseng are attributed to ginsenosides. Ginsenoside Re is one of the active ingredients in ginseng. The present study was carried out to characterize the toxicity of ginsenoside Re after repeated oral administration in Sprague-Dawley rats. MATERIALS AND METHODS: Rats (60 males, 60 females) were administrated ginsenoside Re orally in 0, 38, 113, or 375 mg/kg/day doses for 26 weeks (n=15/group each sex). Clinical signs, mortality, body weights, feed consumption, urinalysis, hematology, serum biochemistry, gross findings, organ weights and histopathology were examined at the end of the test period, as well as after the 4-week recovery period. RESULTS: Ginsenoside Re did not induce death, adverse effects or dose-dependent changes in feed consumption, or body weight gain. Some statistically significant differences were observed in hematological and biochemical parameters, as well as in body weights of rats treated with ginsenoside Re. However, there was no abnormality of any organs noted in both gross and histopathological examinations. CONCLUSIONS: Ginsenoside Re is well tolerated up to a 375 mg/kg/day oral dosage level and non-toxic in both male and female rats.

Subchronic toxicity studies with ginsenoside compound K delivered to dogs via intravenous administration.

Compound K, i.e., 20-O-ß-d-glucopyranosyl-20(S)-protopanaxadiol, is the main metabolite of the protopanaxadiol type of ginseng saponin produced by intestinal bacteria after oral administration of ginseng extract. In the present study, the toxicity of compound K was evaluated in male and female dogs after 90 days continuous intravenous infusion. Beagle dogs were treated with compound K at doses of 6.7, 20 and 60 mg/kg/day, and observed for 90 days followed by recovery periods. Measurements included clinical observations, body weight, food consumption, temperature, electrocardiogram (ECG), hematology, blood chemistry, urinalysis, gross necropsy, organ weight and histopathology. Under the conditions, the clinical condition of the animals, body weights, body weight gains and food consumption were unaffected by compound K administration relative to the control group. Hematology, ECG data and urinalysis parameters were also unaffected. However, the hepatotoxicity was evident from the observation of multiple parameters, including histopathological evaluation of liver tissue upon necropsy as well as large increases in plasma levels of liver enzymes (alanine aminotransferase, ALT, Gamma-glutamyltranspeptidase, γ-GT, alkaline phosphatase,ALP) in groups receiving compound K (20 or 60 mg/kg/day), and this hepatoxicity might be reversible. In addition, the NOAEL of compound K is 6.7 mg/kg/day in this 90 days toxicity study.

Steamed ginseng-leaf components enhance cytotoxic effects on human leukemia HL-60 cells.

Three new dammarane-type glycosides, named ginsenosides SL(1)-SL(3) (1-3), and eleven known compounds (4-14) were isolated from the heat-processed leaves of Panax ginseng. Their structures were elucidated on the basis of extensive chemical and spectroscopic methods. Cytotoxic-activity testing of compounds 1-14 against human leukemia HL-60 cells showed that ginsenosides Rh(3) (11) and Rk(2) (12) exhibited potent effects with IC(50) values of 0.8 and 0.9 microM. In addition, ginsenosides SL(3) (3), 20S-Rg(2) (7), F(4) (10), 20S-Rh(2) (13) displayed strong activity with IC(50) values of 9.0, 9.0, 7.5, and 8.2 microM, respectively. This is the first report on chemical components of the steamed ginseng leaves.

Vascular smooth muscle dysfunction and remodeling induced by ginsenoside Rg3, a bioactive component of ginseng.

Ginseng, one of most well-known herbal medicines, is widely and indiscreetly used among the patients with cardiovascular disorders, raising concern over abuse of this medicine and unwanted effects. In this study, we investigated the effects of ginsenoside Rg3 (Rg3), an active ingredient of ginseng, on vascular contractility and structural integrity to explore its potential vascular toxicity. In isolated rat aorta, Rg3 suppressed the normal agonist-induced contractile response. This suppression persisted even after a rigorous washout. In the endothelium-denuded aortic ring, impairment of vascular contractility by Rg3 was retained, suggesting that vascular smooth muscle was affected. In primary vascular smooth muscle cells, Rg3 abolished agonist-induced Ca(2+) increase, indicating that Ca(2+) regulation was disrupted. Rg3 suppressed the contraction induced by Bay K8644, an L-type Ca(2+) channel activator, whereas store-operated Ca(2+) channel or intracellular Ca(2+) store-mediated contraction was not affected, suggesting that the L-type Ca(2+) channel was selectively impaired by Rg3. These in vitro results were further confirmed in vivo where Rg3 treatment significantly attenuated the agonist-induced pressor response. More importantly, 4-week repeated treatment with Rg3 in normal animals induced eutrophic outward remodeling in the thoracic aorta, that is, it brought about an increased luminal area without changes in the wall area. These results suggest that Rg3 can induce the vascular smooth muscle dysfunction by disturbing Ca(2+) influx from the L-type Ca(2+) channel, ultimately leading to impaired vascular contractility and structural remodeling.

Ginsenoside Rd from Panax notoginseng is cytotoxic towards HeLa cancer cells and induces apoptosis.

The saponin ginsenoside Rd (1), isolated from Panax notoginseng, is used for the treatment of cardiovascular diseases, inflammation, different body pains, trauma, and internal and external bleeding due to injury. In this study, we report that 1 inhibits the cell growth of human cervical cancer (HeLa) cells in a concentration- and time-dependent manner, with an IC(50) value of 150.5+/-0.8 mcirog/ml after 48 h of incubation. The drug-treated cells displayed features of apoptosis, including typical morphological characteristics and formation of DNA ladders, as evident from agarose-gel electrophoresis. Flow-cytometric analysis showed that the cell-cycle distribution of HeLa cells exposed to 1 is characterized by a decrease of the G(0)/G(1)-phase and an increase of the S-phase cells, respectively, in a dose-dependent manner. The apoptotic rate of HeLa cells treated for 48 h with 210 microg/ml of 1 was 35.8%. Further, 1 was found to increase the expression of Bax and to decrease the expression of Bcl-2 proteins, respectively, and to lower the mitochondrial transmembrane potential of HeLa cells. The caspase-3 inhibitor DEVD-CHO (at 2 microM) increased the viability of HeLa cells treated with 1. Taken together, our study suggests that ginsenoside Rd (1) significantly inhibits HeLa cell proliferation, and induces cell apoptosis through down-regulating Bcl-2 expression, up-regulating Bax expression, lowering the mitochondrial transmembrane potential, and activating the caspase-3 pathway. Thus, 1 could serve as a lead to develop novel chemotherapeutic or chemopreventive agents against human cervical cancer.

Effects of ginsenoside Rg1 on postimplantation rat and mouse embryos cultured in vitro.

GRg1 is one of the most important active agents extracted from ginseng. Although there are numerous reports in the medical literature concerning the beneficial effects of GRg1, little is known about its effects on embryonic development. In our study, whole embryo culture was applied to explore the effects of GRg1 on SD rat and ICR mouse embryos during their critical period of organogenesis. All embryos were exposed to different concentrations of GRg1, and scored for their growth and differentiation at the end of the 48-h culture period. Total morphological score decreased significantly at the concentration of GRg1 of 50 microg/ml to mice and at 30 microg/ml to rats. The whole embryonic growth was affected, represented as reduced crown-rump length and head length, and decreased number of somites from 10 microg/ml both in mice and rats. Morphological development was influenced relatively late. To rats, the scores of flexion, forelimb bud and hindlimb bud were significantly reduced at 30 microg/ml, and heart was among the affected organs. However, the effect on heart was not observed in mice, and flexion and limb buds were influenced at 50 microg/ml. These results demonstrate that GRg1 has embryotoxicity during both rat and mouse organogenetic period and rats might be more sensitive than mice. We suggest that pregnant women of first trimester should use ginseng with caution before the exact conclusion is obtained.

Developmental toxicity research of ginsenoside Rb1 using a whole mouse embryo culture model.

BACKGROUND: Ginseng has been widely used around the world for many years. Knowledge is limited, however, on its effects on embryonic development. METHODS: Whole embryo culture was used to explore the developmental toxicity of ginsenoside Rb1 (GRb1) on mouse embryos. All embryos were exposed to different concentrations of GRb1, and scored for their growth and differentiation at the end of the 48-hr culture period. RESULTS: Total morphological score decreased significantly at the concentration of GRb1 of 30 microg/ml and was further reduced at 50 microg/ml. Yolk sac was affected at the lower concentration of 15 microg/ml. Developments of midbrain, forebrain, and optic system were relatively sensitive to GRb1 and were affected at the concentration of 30 microg/ml. Allantois, flexion, branchial arch, and limb buds were affected at 50 microg/ml. At this concentration, the embryonic crown-rump length, head length, and somite number were also reduced significantly compared to the control group. CONCLUSIONS: These results suggest that GRb1 has teratogenic effect during the mouse organogenetic period. We suggest that before more data in humans is available, ginseng should be used with caution by pregnant women in the first trimester.

Embryotoxicity study of ginsenoside Rc and Re in in vitro rat whole embryo culture.

BACKGROUND: Pregnant women commonly consume ginseng. However, there is little data concerning the effects of ginseng on early pregnancy. METHODS: Rat embryos were exposed in vitro to different concentrations of Rc and Re from day 9.5 to day 11.5 after conception. Embryos were scored for growth and differentiation at the end of the culture period. RESULTS: Embryos exposed to 50.0 microg/ml Re had significantly lower median morphological score (29.0 versus 48.0), fewer number of somites (15.0 versus 21.0), and smaller yolk sac diameter (3.5 versus 4.1 mm) and crown-rump length (CRL) (2.9 versus 3.4 mm) compare to control embryos. There was no significant difference between embryos exposed to 5.0 microg/ml Re and control embryos. There was also no difference in the biometric and morphologic parameters among control and embryos exposed to 5.0 and 50.0 microg/ml Rc. CONCLUSION: There is a significant variability in embryotoxic effects of different ginsenosides. Further studies to evaluate the synergistic embryotoxic effects of ginsenosides are warranted.

Mechanistic studies on protopanaxadiol, Rh2, and ginseng (Panax quinquefolius) extract induced cytotoxicity in intestinal Caco-2 cells.

Certain ginsenosides, also known as triterpene glycosides, have been recently reported to have a characteristic effect on cultured intestinal and leukemia cell growth. Ginsenoside aglycones 20(S)-protopanaxadiol (PD), 20(S)-protopanaxatriol (PT), and ginsenoside Rh2 have been identified as having a strong effect on reducing cell viability. Furthermore, ginsenoside Rh2 is thought to be a rare ginsenoside not found in all ginseng products. Rather, Rh2 has been recently reported to be a breakdown product of thermal processing of North American ginseng. In this study, pure ginsenosides PD, PT, Rh2 standards and an enriched Rh2 fraction derived from ginseng leaf were tested in cultured Caco-2 cells for relative cytotoxic potency. PD and Rh2 LC50 were similar after 24 to 72 h, whereas a drop in PT LC50 occurred later at 48 and 72 h. Furthermore, PD and Rh2 affected membrane integrity as indicated by LDH secretion earlier than PT and the enriched Rh2 fraction (P < or = 0.05). Ginsenoside Rh2 showed the greatest (P < or = 0.05) build up of necrotic cells (18.3 +/- 0.1%) at the respective LC50 after 24 h and PD (21.3 +/- 0.3%) showed the largest effect after 44 h of exposure. The effect on apoptotic cells at 44 h of treatment were significantly different (P < or = 0.05) for Rh2 (21 +/- 0.4%), PD (14.6 +/- 0.1%), enriched Rh2 leaf fraction (9.9 +/- 0.6%), and PT (2.3 +/- 0.1%) treatments. Caco-2 caspase-3 activity was different between ginsenoside exposure; Rh2 (10.6 +/- 0.3 nM pNA) had the greatest (P < or = 0.05) activity followed by the enriched Rh2 leaf fraction (8.3 +/- 0.2 nM pNA), PT (7.3 +/- 0.3 nM pNA). The PD (4.8 +/- 0.04 nM pNA) treatment was similar to untreated cells (4.3 +/- 0.05 nM pNA) in caspase-3 activity. These results show variable bioactive response in cultured intestinal cell to specific ginsenosides and an enriched Rh2 North American ginseng extract which may be explained on basis of hydrophobic/hydrophilic balance.

Structure-function relationship exists for ginsenosides in reducing cell proliferation and inducing apoptosis in the human leukemia (THP-1) cell line.

Ginsenosides of the 20(S)-protopanaxadiol and 20(S)-protopanaxatriol classifications including the aglycones, protopanaxadiol (PD), protopanaxatriol (PT), and ginsenosides Rh2 and Rh1 were shown to posses characteristic effects on the proliferation of human leukemia cells (THP-1). A similar efficacy was not apparent for ginsenoside Rg3. The concentrations to inhibit 50% of cells (LC50) for PD, Rh2, PT, and Rh1 were 13, 15, 19, and 210 microg/mL, respectively. PD and PT induced DNA fragmentation at the LC50 after 72 h of treatment, compared to Rh2, Rh1, dexamethasone, and untreated cells. Cell-cycle analysis confirmed apoptosis with PD and PT treatment of THP-1 cells resulting in a buildup of sub-G1 cells after 24, 48, and 72 h of treatment. Rh2 and dexamethasone treatments also increased apoptotic cells after 24 h, whereas Rh1 did not. After 48 and 72 h, Rh2, Rh1, and dexamethasone similarly increased apoptosis, but these effects were significantly (P<0.05) lower than those observed for both PD and PT treatments. Furthermore, treatments that produced the largest buildup of apoptotic cells were also found to have the largest release of lactate dehydrogenase. It can be concluded from these studies that the presence of sugars in PD and PT aglycone structures reduces the potency to induce apoptosis, and alternately alter membrane integrity. These cytotoxic effects were different to THP-1 cells than dexamethasone.