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OBJECTIVE ：To est ablish the method for the determination of 20（S）-protopanaxadiol（PPD）concentration in human plasma. METHODS ：Plasma samples were precipitated with acetonitrile and determined by UPLC-MS/MS ，using finandrogen as internal standard. The determination was performed on Waters ACQUITY UPLC HSS T 3 column with mobile phase consisted of 5 mmol/L ammonium bicarbonate aqueous solution-acetonitrile （gradient elution ）at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃，and sample size was 10 μL. The ion source was electrospray ion source，and negative ion scanning was carried out with multiple reaction monitoring mode . The ion pairs used for quantitative analysis were m/z 459.40→ 375.20（PPD）and m/z 371.30→315.30（internal standard ）. At the same time ，the method was applied to the determination of clinical samples. RESULTS ：The linear range of PPD was 0.25-30.00 ng/mL（r＝0.999 2），and the limit of quantitation was 0.25 ng/mL. RSDs of intra-batch and inter-batch were all lower than 10％，and relative errors （RE）were －14.61％-12.69％. Extraction method and matrix effect did not affect the quantitative determination of PPD. In ginsenoside CK 100 mg group ，ginsenoside CK 200 mg group and ginsenoside CK 300 mg group ，mean cmax of patients with rheumatoid arthritis after oral administration of corresponding drugs were 18.06，30.03，27.00 ng/mL；median tmax were 12.0，6.0，12.0 h；mean AUC 0-t were 622.52，668.15， 1 155.97 ng·h/mL. CONCLUTIONS ：The method for the determination of PPD concentration in human plasma is successfully established. The method is sensitive ，accurate， kq1907011） stable，easy to operate and less plasma consumption. It can be used for the quantitative determination of clinical samples.
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Although the tumor suppressor P53 is known to regulate a broad network of signaling pathways, it is still unclear how certain drugs influence these P53 signaling networks. Here, we used a comprehensive single-cell multiomics view of the effects of ginsenosides on cancer cells. Transcriptome and proteome profiling revealed that the antitumor activity of ginsenosides is closely associated with P53 protein. A miRNA-proteome interaction network revealed that P53 controlled the transcription of at least 38 proteins, and proteome-metabolome profiling analysis revealed that P53 regulated proteins involved in nucleotide metabolism, amino acid metabolism and "Warburg effect". The results of integrative multiomics analysis revealed P53 protein as a potential key target that influences the anti-tumor activity of ginsenosides. Furthermore, by applying affinity mass spectrometry (MS) screening and surface plasmon resonance fragment library screening, we confirmed that 20()-protopanaxatriol directly targeted adjacent regions of the P53 DNA-binding pocket and promoted the stability of P53-DNA interactions, which further induced a series of omics changes.
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Objective: To study the chemical constituents in acid hydrolysates of Panax notoginseng saponins (PNS). Methods: These compounds were separated and purified by column chromatography, and their structures were elucidated based on spectroscopic analyses (HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, HSQC and HMBC). Results: Eighteen compounds were obtained from the acid hydrolysates of PNS and characterized as dammar-25-ene-24-hydroperoxyl-3β,6α,12β,20S-tetraol (1), 6α,12β,20S-trihydroxy- dammarane-24-ene-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (2), 6α,12β,20R-trihydroxy-dammarane-24-ene-3-O-β-D- glucopyranosyl-(1→2)-β-D-glucopyranoside (3), vina-ginsenoside-R8 (4), 24(S)-pseudo-ginsenoside-GQ (5), ginsenoside Rg5 (6), 20 (R)-ginsenoside Rg3 (7), 20(R)-ginsenoside Rk2 (8), 3β,12β-dihydroxy-dammar-(E)-20(22),24-diene-6-O-β-D-xylopyranosyl- (1→2)-β-D-glucopyranoside (9), 20(S)-ginsenoside Rg2 (10), ginsenoside SL1 (11), 20(R)-ginsenoside Rh1 (12), 20(22) E-ginsenoside Rh4 (13), 25-hydroxy-20(R) ginsenoside-Rh1 (14),3β,6α,12β,20(S)-20,25-epoxy-3,12-dihydroxy-dammarane-6-O-β-D-glucopyranoside (15), 20(R)-protopanaxadiol (16), 20(R)-protopanaxatriol (17), and 20(S)-protopanaxatriol (18). Conclusion: Compound 1 is a new triterpen saponin, and compounds 2-5 are isolated from P. notoginseng and acid dydrolysates of PNS for the first time.
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Introduction: This study examined the significance of support groups from the perspective of families for members who were parents of mentally disabled children in their 20s.Methods: We conducted semi-structured interviews with parents who joined support groups for families of mentally disabled young adults. A qualitative and inductive classification method was used to extract categories from the data obtained.Results: Five parents agreed to participate in an interview survey. Ten categories were identified: "talking to others in the same situation", "pleasant places where they feel at ease", "connecting with other families", "sharing the same issues with others", "talking about things other than their children", "learning from each other", "promoting empowerment as a parent", "dealing with their children more easily", "improving family relationships", and "preparation for raising issues within the community".Conclusion: Parents who joined support groups for families of mentally disabled young adults felt that it was important to share problems regarding their children's siblings, the types of housing their children are faced with living in when they become independent, and the types of insurance plans their children can purchase. They also found it important for family support groups to be able to help prepare them to raise issues within the community.
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@#To study the inhibitory effect of gigantol on proliferation, migration and invasion of human osteosarcoma U20S cells and to explore the mechanism. Methods: After being treated with different concentrations (10, 25, 50, 75, 100, 150 µmol/L) of gigantol for 24 and 48 h, the proliferation of U20S cells was detected by CCK-8 assay. Transwell assay was used to detect the effects of 25 µmol/L and 50 µmol/L gigantol on the migration and invasion abilities of U20S cells. The lipopolysaccharide (LPS) was used to induce inflammatory reaction in U20S cells before gigantol treatment; qPCR and WB were used to detect the mRNA and protein expressions of NF-κB (p65), TNF-α, IL-6 and PRL-3, respectively. Results: Different concentrations of gigantol could all inhibit the proliferation of sarcoma U20S cells at different time (P<0.05 or P<0.01). The 25 µmol/L and 50 µmol/L of gigantol could significantly inhibit the migration and invasion of osteosarcoma U20S cells (all P<0.01); at the same time, it could inhibit the protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 (P<0.05 or P<0.01). After LPS induction, the mRNA and protein expressions of NF-κB, TNF-α, IL-6 and PRL-3 in U20S cells were significantly increased (all P<0.01); however, the consequent treatment with gigantol (25 and 50 µmol/L) reversed the effects of LPS on U20S cells obviously (P<0.05 or P<0.01). Conclusion: Gigantol can inhibit the proliferation, migration and invasion of osteosarcoma U20S cells, and its mechanism may be related to the regulation of NF-κB/PRL-3 signaling pathway.
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Objective: To study the minor triterpenoid saponins from the roots of Panax notoginseng, which provided basis for the systematic research, quality control and safety evaluation of P. notoginseng. Methods The compounds were isolated and purified by MCI resin, ODS, along with Preparative-HPLC, and the structures were identified by spectroscopic analysis, and comparing with the pubished literature values. Results: Twelve monomeric compounds isolated from the roots of P. notoginseng, were identified as notoginsenoside P1 (1), notoginsenoside T5 (2), ginsenoside Rk3 (3), ginsenoside Rh4 (4), notoginsenoside T3 (5), 20(S)-protopanaxatriol (6), dammar 20 (21),24-diene-3β,6α,12β-triol (7), ginsenoside Rg3 (8), gypenoside XIII (9), ginsenoside Rk1 (10), ginsenoside Rg5 (11), and 20 (S)-ginsenoside Rh2 (12). Conclusion: Compound 1 is a new dammarane-type triterpenoid saponin
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Objective: To study the chemical constituents of ginsenosides from the flower buds of Panax ginseng. Methods: The compounds were isolated and purified by Diaion HP-20, MCI gel, silica gel, and semi-preparative HPLC. The structures were elucidated based on NMR and MS data. Results: Four compounds were isolated from the extract of P. ginseng flower buds, and identified as 6’-acetyl-ginsenoside F1 (1), 12α-hydroxyl-ginsneoside Rd (2), 20(S)-ginsenoside Rg3 (3), and 5,6-didehydro-20(S)- ginsenoside Rg3 (4). Conclusion: Compound 4 is a novel ginsenoside, compounds 1 and 2 are new natural products.
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@# Objective: To investigate the relationship between long non-coding RNA (lncRNA) HIT and cisplatin (DDP) resistance in osteosarcoma cells and the mechanism related to epithelial-mesenchymal transition (EMT). Methods: 42 pairs of osteosarcoma tissues and corresponding para-cancerous tissues (more than 5 cm away from the edge of cancer tissues) were collected at the Department of Orthopedics, Tianjin Hospital during June 2017 to June 2018. Quantitative Real-time PCR (qRT-PCR) was used to detect the mRNAexpression of HIT and EMT related markers (Snail and E-cadherin) in the collected tissues. The DDP-resistant osteosarcoma U2OS cell line was constructed and human adrenal 293T cell line was used as control. Two sets of siRNA vectors targeting HIT loaded on lentivirus were transfected into cells with DDP-resistance as the interference group A and group B. Meanwhile, the U2OS cell line was transfected with HIT full-length vector and blank vector respectively, as over-expression group and blank group. The DDP 50% inhibitory concentration (IC50) was detected by MTT assay. qRT-PCR was used to detect the mRNA expressions of HIT, Snail and E-cadherin. Western blotting was used to detect the protein expressions of Snail and E-cadherin. RNA binding protein immunoprecipitation (RNAIP) assay was used to clarify the combination of HIT and Snail protein in the U2OS and 293T cells. Results: The mRNAexpressions of HIT and Snail in osteosarcoma tissues were significantly higher than those in para-cancerous tissues, while the mRNA expression of Ecadherin was significantly lower than that in the paracancerous tissues. The mRNA expression of HIT and E-cadherin in osteosarcoma tissues was negatively correlated (all P<0.01). The DDP IC50 in the DDP-resistance group was significantly higher than that in the control group, interference group A and B, and the DDP IC50 in over-expression group was significantly higher than that in blank group (all P<0.01). The expression of HIT in resistance group was significantly higher than that in the control group, and the HIT expressions in interference group A and B were significantly lower than that in DDP-resistance and control group; moreover, the expression of HIT in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The mRNAexpression of Snail in DDPresistance group was significantly higher than that in the control group and interference group A and B, while the mRNA expression of E-cadherin in DDP-resistance group was significantly lower than that in the control group and interference group A and B; and the mRNA expression of E-cadherin in over-expression group was significantly lower than that in blank group. The protein expression of Snail in the DDP-resistance group was significantly higher than that in the control group and interference group A and B,while E-cadherin protein expression was significantly lower; and protein expression of Snail in over-expression group was significantly higher than that in blank group (all P<0.05 or P<0.01). The expression of HIT in the U20S and 293T cells treated by anti-Snail antibody induced by immunomagnetic beads was significantly higher than that in the cells treated by IgG antibody (P<0.01). Conclusion: HIT can promote EMT and cisplatin-resistance in osteosarcoma cells through up-regulation of Snail protein and inhibition of E-cadherin transcription activity.
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Objective To establish the HPLC fingerprint and simultaneous method for the determination of 11 saponins in steamed Panax notoginseng from different origins. Methods Agilent Zorbax SB-C18 (250 mm × 4.6 mm, 5 μm) column was adopted, the mobile phase consisted of acetonitrile-water with gradient elution at the flow rate of 1.0 mL/min (elution program: 0-30 min, 20% acetonitrile; 30-60 min, 20%-45% acetonitrile; 60-88 min, 45%-75% acetonitrile; 88-98 min, 75% acetonitrile; 98-100 min, 75%-20% acetonitrile), and the detection wavelength was 203 nm. The establishment of steamed P. notoginseng HPLC fingerprint, and determination method of notoginsenoside R1, ginsenoside Rg1, Re, Rb1, 20S-Rh1, 20R-Rh1, Rd, Rk3, Rh4, 20S-Rg3, and 20R-Rg3 index components were studied methodologically. The content of 11 saponins in 10 batches was determined. Results The HPLC fingerprint was establish, and thirty common peaks were selected as characteristic peaks of steamed P. notoginseng. The similarities of different samples from 10 areas were 0.941, 0.938, 0.945, 0.951, 0.913, 0.909, 0.920, 0.928, 0.917, and 0.919. In quantitative analysis, eleven saponins were separated well and the average content was 5.274, 20.515, 2.838, 23.651, 3.476, 1.407, 5.239, 1.784, 1.580,0.904, and 0.294 mg/g, respectively. Additionally, all calibration curves showed good linear regression relationship, with correlation indexes of 0.999 7, 0.999 5, 0.999 5, 0.999 7, 0.999 7, 0.999 6, 0.999 7, 0.999 6, 0.999 7, 0.999 7, and 0.999 6; The average recoveries were 101.23%, 98.52%, 97.67%, 99.62%, 98.17%, 98.92%, 99.44%, 99.14%, 100.25%, 98.23%, and 96.89%, with RSDs of 1.35%, 1.58%, 2.44%, 1.05%,1.48%, 1.56%, 0.85%, 2.34%, 2.85%, 1.25%, and 1.08%. Conclusion This method is sensitive, accurate and reproducible. It can be used to provide a reference for the standard and evaluation of quality of steamed P. notoginseng.
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Objective To investigate the chemical constituents of the fruit pedicels of Panax ginseng. Methods The compounds were purified by silica gel column chromatography and preparative reverse-phase high performance liquid chromatography. Their structures were elucidated on the basis of spectroscopic analyses. Results Twenty-eight compounds were isolated from the total extract of the fruit pedicels of P. ginseng. They were identified as 20(S)-protopanaxatriol (1), 20(S)-protopanaxadiol (2), 20(S)-dammar-24- ene-3-one-6α,12β,20-triol (3), (20S,23E)-dammar-23-ene-3β,6α,12β,20,25-pentol (4), (20S,24R)-dammar-26-ene-3β,6α,12β,20,24- tetrol (5), 20(R)-dammar-3-one-6α,12β,20,25-tetrol (6), ginsenoside Rk2 (7), 20(R)-dammar-24(25)-epoxy-3β,6α,12β,20- tetrol (8), ginsenoside CK (9), 20(S)-dammar-3β,6α,12β,20,25-pentol (10), ginsenoside Rh4 (11), 20(S)-dammar-3β,12β,20,25-tetrol (12), 20(S)-dammar-3β,6α,12β,20,24-pentol (13), 20(R)-dammar-3β,6α,12β,20,25-pentol (14), pseudoginsenoside RT5 (15), ginsenoside Rh1 (16), ginsenoside Rh3 (17), 20(S)-dammar-3β,12β,20,25-tetrol-3-O-β-D-glucopyranoside (18), 20(S)-isoginsenoside Rh3 (19), ginsenoside Rg1 (20), ginsenoside Y (21), ginsenoside Rd (22), ginsenoside la (23), 20(S)-dammar-3β,12β,20,25-tetrol-3-O-β-D-glucopyranosyl-(1→2)-O-β-D-glucopyranoside (24), ginsenoside Rg2 (25), notoginsenoside Fe (26), ginsenoside Re (27), and ginsenoside Rb1 (28), respectively. Conclusion Among them, compounds 4-6, 8, 10, 12, 13, and 21 are isolated from P. ginseng for the first time.
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To establish a HPLC-MS/MS method for simultaneous determination and active ingredients screening of pseudoginsenoside RT5, 20(S)-ginsenoside Rh1 and 20(S)-ginsenoside Rh2 by cell membrane chromatography (CMC) in secondary ginsenoside H dripping pills (SGHDP). Methods The samples were separated on Century SIL BDS C18 column (250 mm × 4.6 mm, 5 μm) eluted with 0.2% formic acid aqueous solution-acetonitrile in a gradient mode, and the target compounds were analyzed by positive ion multiple reaction monitoring (MRM) mode, and active ingredients of SGHDP obtained in solid-phase of biomembrane by CMC technology were determined at the same time. Results The linear ranges of pseudoginsenoside RT5, 20(S)-ginsenoside Rh1, and 20(S)-ginsenoside Rh2 were 0.095-0.235, 0.042-0.168, and 0.105-0.419 mg/mL; the extraction recoveries were 99.95%, 100.12%, and 100.06%; and RSD were 1.06%, 0.96%, and 0.91%, respectively. The contents of pseudoginsenoside RT5, 20(S)-ginsenoside Rh1, and 20(S)-ginsenoside Rh2 in SGHDP were 21.24%, 21.42%, and 29.70%, respectively. 20(S)-Ginsenoside Rh2 was the active ingredient obtained by biomembrane using as a new quality control maker for SGHDP. Conclusion The developed method is accurate and reliable for the determination of three ginsenosides in SGHDP, and provides a new reference for quality control of SGHDP. 20(S)-Ginsenoside Rh2 is a immobilization component of red cell membrane, speculated to be the active ingredient of SGHDP, which is in consistent with previous studies on antitumor and antidepression.
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Objective:To evaluate the effect of panax quinquefolium 20s-protopanaxtriolsaponins ( PQTS) on the inflammatory reac-tion after cerebral ischemia/reperfusion injury in rats.Methods: Wistar rats were randomly divided into the sham , the model, PQTS treatment groups respectively at dose of 12.5, 25.0 and 50.0 mg· kg-1 and the positive control group (flunarizine hydrochloride injec-tions, FHI, 2.0 mg· kg-1).All the rats were with intraperitoneal injection once a day for 3 consecutive days.The rats were subjected to 2-hour transient middle cerebral artery occlusion (MCAO) followed by 24-hour reperfusion.The neurological function was scored and the infarct volume was measured after the 24-hour reperfusion.Brain edema was measured by the dry-wet weight.Myeloperox-idase ( MPO) activity was determined by a spectrophotometer .The permeability of blood brain barrier was evaluated by the Evans blue content in brain determined by a spectrophotometer .The mRNA levels of interleukin-1β(IL-1β), interleukin-6 (IL-6) and tumor necrosis fac-tor-α( TNF-α) were evaluated by QPCR test .Moreover , the expression of NF-κBp65 was analyzed using Western blotting .Results:Compared with the model group , PQTS and FHI treatment groups (12.5, 25.0 and 50.0 mg· kg-1 ) and FHI group could improve the neurological function , decrease the infarct size , reduce the brain water content , inhibit the MPO activity and reduce the permeability of blood brain barrier.In addition, PQTS and FHI treatment groups (12.5, 25.0 and 50.0 mg· kg-1 ) also effectively inhibited the in-crease in the mRNA levels of IL-1β, IL-6 and TNF-α, and reduced the protein expression of NF-κBp65 when compared with the model group (P<0.05 or P<0.01).Conclusion:The results indicated that PQTS and FHI can significantly protect brain against cerebral is-chemia/reperfusion injury in rats by anti-inflammatory actions.
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Objective:To explore the intervention effect of proteasome inhibitor MG132 in rats with collagen-induced arthritis(CIA),which resembles human rheumatoid arthritis(RA).Methods:Forty-eight female SD rats were randomly divided into three groups,including blank control group,CIA model group and MG132-treated group.There were sixteen rats in each group.Rats in CIA model group and MG132-treated model group were injected with type Ⅱ collagen to established CIA rats.21 days after the initial immunization,the rats in the MG132-treated model group were injected subcutaneously with 1 mg/kg MG132 once daily for 2 weeks.42 days after the initial immunization,the change of paw-swelling and the arthritis scores were determined.The synovial pathology examination was performed with HE staining.The 20S proteasome activity in synovial tissue was measured by fluorescence substrate assay.The expression of NF-κB/p65,IκBα in synovial tissue were analyzed by Western blot.Results:Proteasome inhibitor MG132 significantly attenuated the severity of arthritis and histopathological changes in CIA rats.Compared with the blank control group,the 20S proteasome activity was increased significantly in the CIA model group(P<0.05),and decreased after injection of MG132.Compared with CIA rats,the expression of NF-κB/p65 significantly decreased in rats treated with MG132(P<0.01).Compared with the blank control group,the expression of IκBα protein decreased in CIA model group.After injected with MG132,the protein was significantly increased(P<0.01).Conclusion:The proteasome inhibitor MG132 may attenuates the severity of arthritis and histopathological changes in CIA rats.These effects may be mediated through the inhibition of NF-κB activity.
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Objective: To study the chemical constituents of transformed products by Fusarium sacchari from the leaves saponin of Panax ginseng. Methods: Transformation products separated by the process of silica gel column, compounds were identified and elucidated by spectral and chemical methods. Results: Ten compounds were isolated from transformed products of Fusarium sacchari from the leaves saponin of P. ginseng and identified as 20 (S)-panaxadiol (1), 20 (S)-protopanoxa-diol (2), 20 (R)-protopanoxa-diol (3), 20 (S)-panaxatriol (4), 20 (S)-protopanaxatriol (5), 20 (R)-protopanaxatriol (6), 20 (S)-protopanoxadiol-20-O-β-D-glucopyranose (7), ginsenoside-F1 (8), ginsenoside Rh1 (9), and ginsenoside Rg1 (10). Conclusion: Compounds 1-10 are the compounds of ginsenosides, isolated from transformed products of F. sacchari from the leaves saponin of P. ginseng for the first time. F. sacchari could transform leaves saponin of P. ginseng to rare antitumor saponin, which is a rare active microbial strain with exploitation value.
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Objective To investigate the chemical constituents of the rhizomes of Ligusticum chuanxiong and discuss the significance of first discovery of ginsenosides from the plant. Methods The compounds were isolated and repeatedly purified by column chromatographies such as macroperous resin, Sephadex LH-20, silica gel, and preparative TLC as well as semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties, NMR, and MS spectral analyses. Results Three ginsenoside compounds were isolated from the n-butanol extracts of rhizomes of L. chuanxiong, and their structures were identified as (20S)-ginsenoside Rh1 (1), (20R)-ginsenoside Rh1 (2), and (20R)-ginsenoside Rg3 (3). Conclusion Ginsenosides are isolated from the genus Ligusticum (Umbelliferae) for the first time, it is of great significance for clarifying pharmacodynamic material basis of the rhizomes of L. chuanxiong. These results also provide the reference data for further verifying the relevance of plant evolution and traditional efficacy between L. chuanxiong and Panax ginseng.
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The 26S proteasome is a 2.5 MDa complex of the protease family members and is central to a vast array of vital cellular processes including cell-cycle progression and antigen presentation. It has been proven to be a target for therapeutic agents in the treatment of cancers and autoimmune diseases. Most inhibitors are designed to target the 20S proteolytic core complex while the efforts to target the 19S regulatory particle subunits are less successful so far. This is, in part, due to the complexity of molecular architecture and poor understanding of the mechanism of this subcomplex. This review attempts to summarize the development of inhibitory molecules that target both the 20S and 19S subunits of the proteasome, especially highlight the recent progress in the proteasome structure and development of the new inhibitors.
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The intracellular calcium ions (Ca) act as second messenger to regulate gene transcription, cell proliferation, migration and death. Accumulating evidences have demonstrated that intracellular Cahomeostasis is altered in cancer cells and the alteration is involved in tumor initiation, angiogenesis, progression and metastasis. Targeting derailed Casignaling for cancer therapy has become an emerging research area. This review summarizes some important Cachannels, transporters and Ca-ATPases, which have been reported to be altered in human cancer patients. It discusses the current research effort toward evaluation of the blockers, inhibitors or regulators for Cachannels/transporters or Ca-ATPase pumps as anti-cancer drugs. This review is also aimed to stimulate interest in, and support for research into the understanding of cellular mechanisms underlying the regulation of Casignaling in different cancer cells, and to search for novel therapies to cure these malignancies by targeting Cachannels or transporters.
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Objective Based on the previous research that the ethanolic extract from traditional Chinese medicine fructus forsythiae (Lianqiao) can obviously inhibit cancer cells in vitro, the article aimed to investigate the anti-proliferation effects of dammar-24-ene-3β-acetate-20S-ol (DM) extracted from fructus forsythiae on gastric cancer cells and its mechanism.Methods MTT assay was used to assess the anti-proliferation effects of DM on gastric cancer cells including SGC-7901, BGC-823, and MKN-45 in vitro.There were MKN-45 control group and its low dose and high dose groups, BGC-823 control group and its low dose and high dose groups, SGC-7901 control group and its low dose and high dose groups in the experiment.Flow cytometry was used to analyze the cell apoptosis rate.Cellquest software was used to analyze the results and record the ratio of cells at different cycles.DCFH-DA probe was applied to detect the ROS levels of blank control group, docetaxol group and DM group.The reaction system of microtubule assembly test was set with 10?mol/L docetaxol, 50 or 100 μmol/L DM final density and no medicine in blank control group.The readings of UV spectrophotometer were recorded.Microtubule assembly assay and microtubule immunofluorescence staining were applied to investigate the effects of DM on microtubule system.Results The inhibition ratio of 50 μg/L DM on the proliferation three gastric cell lines were all above 80%, with IC50s of MKN-45 11.72±1.35 μg/mL, BGC-823 17.19±0.82 μg/mL, SGC-7901 7.55±0.79 μg/mL.8 days′ low density culturing at 48 hours after 2 μg/mL DM treatment, compared with control group, the number of cell clones significantly reduced without much change in clone size, while 48 hours after 10 μg/mL DM treatment, besides a few clones of BGC-823, there were just several megascopic clones of SGC-7901 and MKN-45.In comparison with apoptotic cell ratio in MKN-45 control group[(21.1±2.5)%], its low dose group and high dose group resulted in significant rise of apoptotic cell ratio[(25.1±1.3)% and (55.2±2.3)%] (P0.05).In comparison with MKN-45 control group, the ratio of cells at S phase decreased in its low dose group[(14.5±2.7)% vs (12.3±3.3)%,P>0.05].In comparison with BGC-823 control group, the ratio of cells at S phase increased in its low dose group[(12.2±5.4)% vs (20.2±2.1)%,P<0.05].In comparison with SGC-7901 control group, the ratio of cells at S phase increased in its low dose group[(21.5±3.8)% vs (31.3±2.6)%,P<0.05].From the detection of intracellular active oxygen after DM treatment, dose-dependent ROS level increased in all three cell lines 48 hours after 10μg/mL and 50μg/mL DM treatment.From the results of microtubule immunofluorescence staining, 48 hours after the treatment of IC50 docetaxol and 10μg/mL DM, the fluorescence signals were in local concentration and disorder.Conclusion Dammar-24-ene-3β-acetate-20S-ol demonstrated anti-proliferation effects due to the apoptosis induced by cell cycle arrest at S phase.
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Objective To explore the excretion of the 20 (S)-protopanaxatriol (PPT) and its metabolites ocotillol type epimers (M1 and M2) in urine, feces samples and the excretion of Ml and M2 in bile samples. Methods The concentration of PPT, Ml and M2 in urine, feces samples and the concentration of Ml and M2 in bile samples were determined by the LC-MS/MS methods with or without the hydrolization by β-glucuronidase. Results After intragastric(ig) administration of PPT, the cumulative excretion rate for 72 h of PPT, Ml and M2 in feces were 14.88%, 1.34% and 0.084%, respectively. With the hydrolization by β-glucuronidase, the cumulative excretion rate for 72 h of PPT, Ml and M2 in feces were 14.77%, 1.36% and 0.085%, respectively. However, the epimers and PPT were hardly detected in urine. After ig administration of M1 or M2, the accumulation excretion rate were 4.41% for M1 and 47.2% for M2 in feces, while both epimers were hardly detected in urine. After ig administration of M1 or M2, the 36 h cumulative biliary excretion rate was 3.01% for M1, and only 0.068% for M2. The 36 h cumulative biliary excretion rate of M1 was 8.80% after intravenous administration, while only 1.24% for M2. Conclusion After ig administration of PPT, a small amount of PPT and its metabolites (Ml, M2) are excreted by the feces but little via urine, and there are stereoselectivity differences in biliary excretion between M1 and M2.
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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.