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@#In order to reveal the intestinal absorption mechanism of saikosaponin d (SSd) in vitro and in vivo, the current research investigated the effects of different experimental conditions (time, concentration, temperature, pH, intestinal segments), transporter inhibitors, paracellular pathway enhancer, metabolic enzyme inhibitors on the intestinal absorption of SSd, in Caco-2 monolayers and a single pass perfusion model in rats.The results showed that the apparent permeability coefficient (Papp) and effective permeability coefficient (Peff) of SSd were 4.75 × 10-7 - 6.38 × 10-7 cm/s and 0.19 × 10-4- 0.27 × 10-4 cm/s, respectively, indicating that it was a low permeability compound, and that the transmembrane transport of SSd was concentration-dependent (0.5-5 μmol/L) and time-dependent (0-180 min).Ileum was the main absorption site for SSd. Experimental results based on Caco-2 monolayers showed that the P-gp inhibitor and paracellular permeability enhancer significantly increased the absorption of SSd (P < 0.05), which was consistent with the results obtained in rats. Inhibitors of OATPs and OCTs showed different results in vitro and in vivo, which may be related to the lower expression of them in jejunum.In summary, the intestinal absorption of SSd occurs through a carrier-mediated and energy-dependent transport, as well as passive diffusion, and P-glycoprotein plays an important role in the active transport of SSd.
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Objective: To investigate the cell death-inducing effect of saikosaponin-d (SSD) on human hepatoma Hep3B cells and its potential mechanism. Methods: Hepatoma Hep3B cells were divided into five groups: blank control group, DMSO (vehicle) group, and three SSD treatment groups treated with various doses of SSD (5, 10, and 15 μmol/L). MTT assay was used to evaluate cell viability. Flow cytometer was employed to quantitatively detect the percentage of dead cells after Annexin V-FITC/PI double staining. Apoptosis was detected morphologically after Hoechst 33258 staining. The activity of caspase-3 apoptotic protease was determined by spectrophotometry. Cell morphologic changes were observed with an inverted microscope. Western blotting and Real-time PCR were employed to evaluate the expression levels of C/EBP homology (CHOP) protein and mRNA, respectively. Results: MTT assay showed that SSD inhibited the viability of human hepatoma Hep3B cells in concentration- and time-dependent manners. Sinomenine hydrochloride induced the death of Hep3B cells in a concentration-dependent manner indicated by flow cytometry. After staining with Hoechst 33258, the nuclei of SSD-treated cells showed nucleosomal agglutination, nucleosomal shrinkage and fragmentation under the fluorescence microscope, which are the characteristics of apoptotic cells. SSD significantly activated the key apoptotic executor caspase-3. The occurrence of paraptosis, characterized by extensive cytoplasmic vacuoles, was observed in SSD-treated cells under an inverted microscope. The pretreatment of a pancaspase inhibitor Z-VAD-FMK completely inhibited caspase-3 activity triggered by SSD, but only partially suppressed cell death and could not reduce the cytoplasmic vacuolation in SSD-treated cells. The protein and mRNA expressions of CHOP, a stress-inducible molecule, were upregulated by SSD, which could not be inhibited by Z-VAD-FMK. Conclusion: SSD can simultaneously induce caspase-dependent apoptosis and caspase-independent paraptosis in human hepatoma Hep3B cells. The upregulated expression of CHOP may be the mechanism involved in SSD-induced paraptosis.
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Objective: To explore the effects of Saikosaponin d (SS-d) on autoimmune hepatitis (AIH) by observing the expression changes of some differentially expressed genes screened with the Agilent-085631 gene chip in the liver of AIH mice. Methods: Forty healthy male SPF C57BL/6 mice were divided into chip group (n=8) and SS-d treatment group (n=32). The mice in the chip group were divided into the normal group and the model group [concanavalin A (Con A) was administered to the tail vein at a dose of 15 mg/kg] (both n=4). The mice were sacrificed after 12 h. The differentially expressed genes of liver were screened, some of which were verified by qRT-PCR. The SS-d treatment group was further divided into the normal group, the model group (treatment was the same with the chip group), SS-d low-dose group and SS-d high-dose group [according to the literature and pre-experiment results, 2.5 and 5.0 mg/kg dose of intraperitoneal injection of SS-d were given respectively, and 15 mg/kg of Con A was administered to the tail vein 8 h later] (all n=8). After 12 h, total venous blood, liver total protein and total RNA of mice were collected. The levels of serum glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), interleukin (IL)-10 and IL-17 were detected by ELISA, and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) was detected by Western blotting. qRT-PCR technology was used to detect the mRNA levels of IL-10, IL-17 and CTLA-4. Results: A total of 11 512 differentially expressed genes were screened (up 5 189, down 6 323), which were related to 138 signal pathways. The qRT-PCR results of IL-10, IL-17 and CTLA-4 gene were consistent with the results of chip screening. Compared with the normal group, the serum levels of GPT and GOT in the model group increased, IL-17 mRNA level increased, IL-10 mRNA and CTLA-4 mRNA levels decreased, the content of serum IL-17 increased, the content of serum IL-10 decreased, and the level of CTLA-4 protein expression in the liver tissues decreased. Compared with the model group, the serum GPT and GOT levels of SS-d in the low-dose and high-dose groups decreased, IL-17 mRNA level decreased, the levels of IL-10 mRNA and CTLA-4 mRNA increased, the content of serum IL-17 decreased, the content of serum IL-10 increased, and the level of CTLA-4 protein expression in the liver tissue increased. Conclusion: Multiple signaling pathways are involved in the pathogenesis of AIH, and SS-d can alleviate the liver inflammation in AIH mice by regulating the expression of IL-10, CTLA-4, and IL-17.
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Objective: To predict the efficacy components and key targets of Chaihu Guizhi Ganjiang Decoction (CGGD) in the intervention of novel coronavirus pneumonia in the cold-dampness obstructing lungs in early stage, and clarify its mechanism. Methods: The novel coronavirus pneumonia TCM stage, clinical manifestations and the function of CGGD were analyzed by literature mining and clinical reports. TCMSP database was used to screen potential active components and related targets in CGGD. PubMed database was used to screen pneumonia, cough and fever related targets. With the help of Cytoscape software, a “drug-disease-target” visual network diagram and protein interaction network were constructed, and GO and pathway enrichment analysis of key targets was performed through the STRING database. The active ingredients were molecularly docked with SARS-CoV-2 3CL hydrolase protein and ACE2 by AutoDock Vina. Results: The analysis of the relationship between prescriptions and syndromes showed that CGGD could play warm-yang scattered cold, resolve dampness, clear stagnation and heat, and open up membrane’s power to intervene in early cold-dampness lung type COVID-19. Through screening, the therapeutic effects of CGGD were mainly in 156 chemical components acting on 159 related targets. The core 27 genes predicted and analyzed included EGFR, TP53, YWHAZ, HSP90AB1, PIK3R1, GRB2, etc. GO and pathway analysis showed that CGGD was mainly involved in biological processes such as cell regulation and immune system related pathways to play a therapeutic role. The 10 core components were molecularly docked, saikosaponin A, saikosaponin D, and peroxyergosterol in CGGD had good affinity with 3CL hydrolase protein and ACE2. Conclusion: Using network pharmacology and molecular docking technology to predict that CGGD can be used for the treatment of novel coronavirus pneumonia with symptom of cold-dampness obstructing lungs in early stage, potential antiviral ingredients contained in prescription of CGGD, can play a therapeutic role in the treatment of new type of coronavirus pneumonia in the early stage by regulating the immune system. It explains the characteristics of “multi-component-multi-target-multi-disease” of Chinese materia medica, and provides theoretical basis for clinical rational use of medicines.
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OBJECTIVE： To establish a method for simultaneous determination of 6 components in Chaihuang tablets， such as baicalin， wogonoside， baicalein， wogonin， saikosaponin a and saikosaponin d in Chaihuang tablets. METHODS： HPLC-DAD method was used to detect 3 batches of Chaihuang tablets from same manufacturers. The determination was performed on Agilent Eclipse XDB-C18 column with mobile phase consisted of acetonitrile-triethylamine phosphate aqueous solution （pH adjusted to 7.0， gradient elution） at flow rate of 1.0 mL/min. The detection wavelengths were set at 210 nm （saikosaponin a， saikosaponin d） and 277 nm （baicalin， wogonoside， baicalein， wogonin）. The column temperature was 30 ℃， and sample size was 5 μL. RESULTS： The linear ranges of baicalin， wogonoside， baicalein， wogonin， saikosaponin a and saikosaponin d were 0.379 5-7.590 4 μg， 0.082 96-1.659 2 μg， 0.039 39-0.787 8 μg， 0.040 72-0.814 4 μg， 0.040 45-0.809 0 μg， 0.038 63-0.772 6 μg （all r≥0.999 3）， respectively. The limits of detection were 0.008， 0.007， 0.005， 0.005， 0.020 and 0.018 μg/mL. The limits of quantitation were 0.025， 0.022， 0.015， 0.015， 0.060， 0.054 μg/mL. RSDs of precision， reproducibility and stability tests （48 h） were all lower than 1.5％ （n＝6）. Average recoveries were 98.46％， 97.06％， 100.90％， 96.13％， 96.91％， 96.57％ （RSD＜2.0％， n＝6）. CONCLUSIONS： Established method is simple， accurate and reproducible for 6 components in Chaihuang tablets， and can be used for quality control of the tablet.
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Objective: The fingerprint of Bupleurum chinense roots was developed with ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UPLC/Q-TOF-MS),the main chromatographic peaks were preliminary identified,and combined with principal component analysis(PCA) pattern recognition method to evaluate the quality of this herb from different origins. Method: The chromatographic separation was performed on a ZORBAX Eclipse Plus C18 column(2.1 mm×100 mm,1.8 μm) with a gradient elution of acetonitrile-0.1%formic acid aqueous solution.The mass spectrometer equipped with electrospray ionization(ESI) was used as detector and operated under the negative ion mode.Taking mass spectrometry data processing software of PeakView 1.2 and metabolomics analysis software of MarkerView 1.2.1,the different origins of B. chinense roots were analyzed by PCA. Result: The fingerprint of B. chinense roots was established within 35 min by UPLC/Q-TOF-MS.The samples from different origins were apparently classified by PCA.Eight compounds with significant differences were screened out,and the structures of three of them were identified as 3″-O-acetyl saikosaponin A,3″-O-acetyl saikosaponin D,6″-O-acetyl saikosaponin D. Conclusion: UPLC/Q-TOF-MS can be used for the rapid identification of fingerprint of B. chinense roots from different origins.IBM SPSS Statistics 21.0 and PCA can comprehensively distinguish the differences of chemical components in B. chinense roots from different origins and can be used to evaluate the quality of this herb.
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Objective: To investigate the mechanisms of effects of Chaihu Guizhi Ganjiang Decoction in the treatment of insomnia by using network pharmacology methods. Methods: TCMSP and TCMID were used to lock the targets of seven herbs in Chaihu Guizhi Ganjiang Decoction. TTD, DrugBank, and PubMed were used to search targets of insomnia and construct a “disease-prescription-target” network. STRING and Cytoscape were used to perform enrichment analysis and clarify the mechanism of core targets in the network. Results: The PPI network of Chaihu Guizhi Ganjiang Decoction contained 640 targets and the PPI network of insomnia included 175 targets. A total of 29 core targets and 80 interactions were found after enrichment analysis between two PPI networks. After GO enrichment analysis and KEGG pathway analysis of 29 key targets, we found that 171 active ingredients in Chaihu Guizhi Ganjiang Decoction such as saikosaponin a, saikosaponin d, quercetin, calcium carbonate, 6-gingerol, kaempferol, and wogonin, which played a role in the treatment of insomnia mainly through 29 core targets such as CACNA1C, GABRA1, GABRA2, GABRB3, GABRA3, with biological processes such as target and synaptic signaling, regulation of membrane potential, G-protein coupled receptor signaling pathway, and molecular functions such as neurotransmitter receptor activity, ion-gated channel activity, GABA-A receptor activity, and functional pathways composed by plasmalemma, synapse, and other cells such as neural active ligand-receptor interaction, retrograde endogenous cannabinoid signal transduction, and serotoninergic synapses. Conclusion: The pharmacological substance basis for the treatment of insomnia was composed of 171 active ingredients such as saikosaponin a and saikosaponin d. The efficacy network of “soothing liver and invigorating spleen, regulating yin and yang” was constituted by several pathways like the neural active ligand-receptor interaction and 29 targets such as CACNA1C. Our results provide network pharmacological evidence for clinical rational use of Chaihu Guizhi Ganjiang Decoction for insomnia.
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OBJECTIVE: To discuss the effect and mechanisms of saikosaponin D (SSD) on apoptosis and autophagy of human hepatocellular carcinoma cells. METHODS: The proliferation of cells in treatment of SSD was detected by MTT. The autophagosome was detected by GFP fluorescence staining. The apoptosis of hepatoma cells were detected by flow cytometry. The expressions of LC3-Ⅱ, p-S6K1 and Beclin 1 in human hepatocellular carcinoma cells was detected by Western blot. RESULTS: The proliferations of PLC, MHCC-97H, SMMC-7721 and Huh7 cells with SSD intervention for 48 h were dose-independent inhibited. The IC50 of PLC, MHCC-97H, SMMC-7721 and Huh7 cells were 51.48, 46.19, 39.87, 48.95 μmol•L-1, respectively. After SSD treatment, the number of autophagosome of PLC, MHCC-97H, SMMC-7721 and Huh7 was significantly more than the normal control group (P<0.05). The apoptosis rate of PLC, MHCC-97H, SMMC-7721 and Huh7 cells induced by SSD+3-MA or SSD+z-DEVD-fmk was less than the SSD treated cells (P<0.05). The expressions of LC3-Ⅱ and phosphorylated S6K1 protein (p-S6K1) in PLC, MHCC-97H, SMMC-7721 and Huh7 cells induced by SSD were more than the normal cells, while the expression of Beclin 1 protein decreased. CONCLUSION: It's suggested that SSD increase the autophagy and apoptosis of human hepatoma cells by regulating the mTORC signaling pathway.
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Objective To study the effects of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, saikosaponin a, and saikosaponin d in Ginseng Radix et Rhizoma-Bupleuri Radix (GRRBR) herb pair and its preparations by using quantitative analysis of multi-components by single-marker (QAMS). Methods On the basis of ginsenoside Rg1, the relative correction factors between the ginsenoside Rg1 and the other four saponins were established, and then the contents of the other four saponins were calculated. At the same time, the contents of the five components were determined by external standard method and compared with those evaluated by QAMS. The relative retention time was determined by different chromatographic columns. It could be considered that QAMS was feasible and accurate in the determination of saponins in GRRBR herb pair. Results A quantitative control method of five kinds of saponins was established, and the methodological results were good. The results showed that there was no significant difference in the contents of five kinds of saponins in GRRBR herb pair, Xiaochaihu Decoction, and Kaixin Jieyu Prescription between QAMS group and external standard method group. Conclusion QAMS is suitable for the determination of saponin in GRRBR herb pair, which can be used as a reference for the determination of its effective components and the establishment of compound quality control method.
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Objective:To research the best processing method for Bupleurum chinense DC.by orthogonal tests.Methods:With the contents of saikosaponin a and saikosaponin d as the indices,the L9(34) orthogonal table was used to study three factors including the amount of wheat bran,pot temperature before heating and processing time.The orthogonal design was applied to study the processing technology of Bupleurum chinense DC.fried with wheat bran.Results:The best processing method was as follows:100 g Bupleurum chinense DC.was mixed with 10 g wheat bran and fried at 290 ℃ for 80 seconds.Conclusion:The optimized processing technology is reasonable,reliable and highly reproducible,which provide reference for the processing of Bupleurum chinense DC.with wheat bran.
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Objective: To evaluate the quality of Radix bupleuri from different producing areas and make a comparison by using grey correlation analysis(GRA)and principal components analysis(PCA).Methods: Fifteen samples of Radix bupleuri were collected from different producing areas.The percentage of total ash, acid-insoluble ash, extract, saikosaponin a and saikosaponin d of Radix bupleuri from different habitats were determined.The grey correlation quality evaluation model was built for Radix bupleuri from different producing areas and the results were compared with those of PCA.Results: According to the grey correlation analysis on components in the different batches of Radix bupleuri , the samples from Shaanxi province were the most highly correlated with the optimal reference sequence (0.79), which suggested the best quality.Radix bupleuri Fructus from Hebei province was the most poorly correlated with the optimal reference sequence (0.23), which showed the worst quality.The results were basically identical with those of PCA, indicating good repeatability and stability.Conclusion: Grey incidence degree method and the established Radix bupleuri quality evaluation models can be used to evaluate the quality of Radix bupleuri .
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Objective: To compare the contents of saikosaponin a, d and total flavonoids in different parts of Hollow bupleurum to provide reference for the clinical use of medicinal parts.Methods: The determination was performed on a SHIMADZU Inertsil C18(250 mm×4.6 mm,5 μm)column.The mobile phase consisted of acetonitrile and water with the ratio of 40∶60, and the flow rate was 1.0 ml·min-1.The detection wavelength was 210nm, the column temperature was 40℃, and the injection volume was 10 μl.A colorimetric method was used to detect total flavonoids with the detection wavelength of 500 nm.Results: The calibration curve of Saikosaponin a, d and total flavonoids showed a good linear relationship respectively over the range of 0.21-1.26 mg·ml-1(r=0.999 6),0.25-1.51 mg·ml-1(r=0.9997) and 4.00-25.00 μg·ml-1(r=0.999 6).The average recovery was 99.27%(RSD=2.15%, n=6),99.4%(RSD=2.14%,n=6)and 99.03%(RSD=1.34%,n=6), respectively.The content of saikosaponin a and d respectively was 0.31% and 0.50% in the root, while that in the other parts was low.The content of total flavonoids was as high as 8.48% in flowers, and that in leaves, stem and root reduced in turn.Conclusion: The aerial parts of Hollow bupleurum are rich in flavonoids, and the content of saikosaponin in root is higher, therefore, the whole plant with root is more reasonable in the clinical use.
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Objective To develop an HPLC method for simultaneous determination of contents of asiaticoside, tetrahydropulmatine and saikosaponin d in Shenji Huwei Granules. Methods The analysis was performed on a R&C C18 column (250 mm × 4.6 mm, 5 μm) by using the mobile phase of acetonitrile (A) - phosphate buffer (which used potassium dihydrogen phosphate 8.34 g, potassium phosphate 0.87 g dissolved by 1000 mL water) with gradient elution (0–15 min, 20％A; 15–30 min, 20％→40％A; 30–42 min, 40％A; 42–45 min, 40％→48％A; 45–50 min, 48％A; 50–70 min, 48％→50％A). The flow rate was 1.0 mL/min; the detection wavelength was set at 210 nm; the column temperature was maintained at 30 ℃. Results Asiaticoside, tetrahydropulmatine and saikosaponin d were in the linear ranges among 0.173–2.770 μg (r=0.9999), 0.021–1.320 μg (r=0.9992), 0.151–9.660 μg (r=0.9993), respectively. The average recovery rates of asiaticoside, tetrahydropulmatine, saikosaponin d were 96.25％, 97.02％, and 97.84％, respectively, and RSD were 2.31％, 4.51％, 1.87％, respectively. Conclusion This method is simple, with good separation effect and strong specificity, and can be used for simultaneous determination of contents of asiaticoside, tetrahydropulmatine and saikosaponin d in Shenji Huwei Granules, which provides references for perfection of quality control of Shenji Huwei Granules.
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ABSTRACT:Objective To observe the influence of saikosaponin-d (SSd)on the proliferation and the function of autophagy of human hepatocellular carcinoma (HCC)cell line SMMC-7721 to explore the possible mechanisms. Methods SMMC-7721 was cultured invitro and then treated with SSd of various concentrations (5.0,7.5,10.0, 12.5,15.0 and 17.5 mg/L)for 24,48 and 72 h.We used MTT to detect cell proliferation,selected the optimal concentration and time,and detected the expressions of BECN1 at mRNA and protein levels by PCR and Western blot.Results The inhibition rate of the proliferation of SMMC-7721 cell line increased with the increase of the concentration of SSd,and the highest inhibition rate (60%)appeared when the concentration reached 12.5 mg/L. The expression of BECN1 in the group with SSd was obviously higher than that in the control group (P<0.05). 3-MA decreased not only the expressions of BECN1 at mRNA and protein levels but also the expression of BECN1 when used in conjunction with SSd.Conclusion The inhibiting function of SSd on SMMC-7721 presents a dependency between drug concentration and function time,basically in line with the drug dose-effect relationship. SSd induces the occurrence of autophagic cell death through up-regulating the expression of BECN1 ,thus inhibiting the proliferation of SMMC-7 7 2 1 .
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Objective To observe the effect of saikosaponin-d on transcriptional activation of estrogen receptor in rat hepatic stellate cells(HSC-T6), and to explore its pharmacological mechanism. Methods The rat HSC-T6 were cultured in vitro for the study. After transient transfection of HSC-T6 with the ER-specific reporter gene ERE-tk-Luc by liposome, the effect of saikosaponin-d and estradiol on luciferase activity was observed by Dual-Luciferase Reporter Assay System under the conditions of various drug concentrations, various treatment time and addition of estrogen receptor inhibitor ICI182. 780. Results When the concentrations of saikosaponin-d and estradiol were in the range of 0.01-5 μmol/L, 0.01-1 μmol/L respectively, luciferase activity was increased in a dose-dependent manner . Luciferase activity arrived the highest when saikosaponin-d concentration was 5 μmol/L and estradiol concentration was 1 μmol/L, but the effect was weakening when estradiol concentration reached 5μmol/L. In respect of the effect of treatment time, when HSC-T6 were separately treated with 5μmol/L of saikosaponin-d and 1 μmol/L of estradiol for 24 h, the luciferase activity was the highest. And ICI182.780 could significantly inhibit the induction of saikosaponin-d and estradiol for luciferase activity. Conclusion Saikosaponin-d has an effect on promoting the transcriptional activation of estrogen receptor in HSC-T6.
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@#This study was to investigate the effect and mechanism of saikosaponin D(SSD)combined with oxaliplatin on nude mice bearing human lung carcinoma A549 cells. The A549 cell-bearing nude mice model was established and then the optimal dosage of SSD for intragastric administration was valued by tumor inhibitory ratio in vivo. The effect of SSD combined with oxaliplatin on tumor growth in A549 cells-bearing nude mice was observed. The apoptosis was detected by TUNEL method, the concentration of prostaglandin E2(PGE2)in plasma was checked by ELISA method, and the expression of COX-2 in tumor was analyzed by Western blot. The result showed that SSD exerted best tumor inhibitory effect(40. 96%)at the dosage of 1. 0 mg/kg. SSD combined with oxaliplatin group induced better apoptosis effect in A549 cells-bearing nude mice than those of SSD group and oxaliplatin group, respectively. The concentration of PGE2 and the expression of COX-2 in model group were increased markedly, while decreased significantly in SSD, oxaliplatin or SSD combined with oxaliplatin treated groups(P< 0. 05), and the concentration of PGE2 and the expression of COX-2 in SSD combined with oxaliplatin group were significantly lower than those treated by SSD or oxaliplatin alone(P< 0. 01). In conclusion, SSD combined with oxaliplatin could exert synergistic effect to induce apoptosis of A549 cells in nude mice, which may achieved by down-regulation of the expression of COX-2.
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Objective: To evaluate the effect of different stem branches of Bupleurum chinense on saikosaponins content in roots and its root yield. Methods: The contents of saikosaponin a, saikosaponin d, and total saikosaponins in the roots of B. chinense were determined by HPLC and UV-vis spectrophotometry. And parameters of growth characteristics were analyzed. Results: The contents of saikosaponin a, saikosaponin d, and total saikosaponins in the roots of B. chinense and its root yield were the highest when the B. chinense stem had no branch or two branches. And with the increase of stem branch's numbers, the content of saikosaponins in the roots of B. chinense and its root yield decreased. Conclusion: Different stem branches have a significant effect on the content of saikosaponins in the roots of B. chinense and its root yield.
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Objective To investigate the anticancer effects and detailed mechanisms of Saikosaponin D (SSD) in human hepatoma HepG2 cells. Methods Cell proliferation and apoptosis were tested by MTT assay and Annexin-V/PI assay respectively. The expressions of CCAAT enhancer binding protein β(C/EBPβ) and p53 were detected by RT-PCR and Western blotting. Results SSD inhibited cell proliferation in a dose-dependent manner and induced apoptosis at the concentration of 5.0mg/L. SSD significantly increased the mRNA and protein levels of C/EBPβ and p53 in a dose-dependent manner. Conclusion SSD exerts its anticancer effect by inhibiting cell proliferation and inducing apoptosis partly through C/EBPβ-p53 signal pathway in HepG2 cells.
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OBJECTIVE:To investigate the effects of saikosaponin-d(SSd) on tissue plasminogen activator(TPA),plasminogen activator inhibitor(PAI),malonaldehyde(MDA) and NO in rats with liver fibrosis induced by dimethylnitrosamine(DMN).METHODS:Eighteen SD healthy rats were randomized to control group(NS ip qd for 4 weeks),model group(10mg? kg-1 DMN ip 3 times per week for 4 weeks) and SSd-treated group(10mg? kg-1 DMN ip 3 times per week + SSd 1.8 mg? kg-1 ip for 28 consecutive days).All rats were killed 1h after the last time of administration,blood samples were taken from abdominal aorta and liver samples were taken for the observation of pathology and detection of indices of TPA,PAI,MDA and NO etc.RESULTS:SSd could lessen the degree of liver fibrosis and improve the fibrinolytic activities of TPA and PAI,meanwhile,it showed clearance effect on MDA and marked protective effect on hepatic cells.There were significant differences between SSd-treated group and the model group.CONCLUSION:SSd exhibited protective function on experimental hepatic fibrosis in rats,which may be attributed to the improving of fibrinolytic activity,eliminating of lipid peroxidation and enhancing of NO level.
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AIM To investigate the pharmacokinetic characters of saikosaponin d in Wuling Capsules in mouse. METHODS Wuling Capsules were given by irrigated (ig) administered to mouse, the blood plasma was collected at different time and the sample solution was prepared by extract adding amine sulfate and the mix fluid of acetonitrile-methyl alcohol (4∶ 1), the content of saikosaponin d was detected by RP-HPLC. The pharmacokinetic parameter was fitted by SIP7 software. RESULTS After ig administered the Wuling Capsules (5.5 g/kg) to mouse, the blood plasma concentration-time course fitted well to one compartment model with the 1st order absorption. The pharmacokinetic parameters of the first absoption in mouse was as following: Ka=2.34_ 1/h ,Ke=0.08_ 1/h ,T_ max =1.45_h,C_ max =83.81_ ?g/ml ,T_ 1/2Ke =7.9 h; and that of the second absorption was as following Ka=1.32_ 1/h ,Ke=0.08_ 1/h ,T_ max =6.48h,C_ Max =119.63_ ?g/ml , T_ 1/2 Ke =7.76 h. CONCLUSION The boipharmaceutics characters of saikosaponin d of Wuling Capsules were absorption slow and elimination was also slow.