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ObjectiveTo investigate the inhibitory effects and mechanism of the compound Phyllanthus urinaria Ⅱ （CPU Ⅱ）on the growth of transplanted hepatocellular carcinoma Hep3B2.1-7 （Short for Hep3R） cells in nude mice. MethodAfter the establishment of a xenograft model of hepatocellular carcinoma Hep3B cells in mice， the model mice were randomly divided into a model group， a high-dose CPU Ⅱ group （57.5 g·kg-1）， a low-dose CPU Ⅱ group （28.75 g·kg-1）， and a 5-fluorouracil （5-FU） group （0.025 g·kg-1）， with eight mice in each group. The mice in the high- and low-dose CPU Ⅱ groups were treated with drugs by gavage， once per day， and those in the model group were treated with the same volume of normal saline. The mice in the 5-FU group were treated by 5-FU by intraperitoneal injection， once every other day. After 28 days of administration， mice were sacrificed， and transplanted tumors were collected. Immunohistochemistry （IHC） was used to detect the expression of proliferating cell nuclear antigen （PCNA） of tumor tissues. Terminal-deoxynucleotidyl transferase-mediated nick end labeling （TUNEL） was used to detect cell apoptosis of tumor tissues. The mRNA expression of miR-122 and insulin-like growth factor 1 receptor （IGF-1R） in tumor tissues was detected by Real-time quantitative PCR （Real-time PCR）. The protein expression of CCAAT/enhancer-binding protein α （C/EBPα）， hepatocyte nuclear factor-4α （HNF-4α）， and IGF-1R in tumor tissues was detected by Western blot. ResultThe tumor suppression rates of the high- and low-dose CPU Ⅱ groups and the 5-FU group were 74.90%， 63.62%， and 64.15%， respectively. Compared with the model group， the CPU Ⅱ groups and the 5-FU group showed reduced weight （P<0.01） and volume of tumors （P<0.01）， decreased PCNA positive cells， shallow staining， increased apoptosis cells of transplanted tumor tissues （P<0.05， P<0.01）， increased expression of mRNA expression of miR-122 （P<0.01）， down-regulated mRNA expression of IGF-1R （P<0.01）， and up-regulated protein expression of C/EBPα and HNF-4α in nude mouse transplanted tumor tissues （P<0.01）. The expression of IGF-1R protein in the high-dose CPU Ⅱ group was down-regulated （P<0.05）. Compared with the low-dose CPU Ⅱ group， the high-dose CPU Ⅱ group showed increased apoptotic cells （P<0.01）， up-regulated mRNA expression of miR-122 （P<0.01）， and increased expression of C/EBPα and HNF-4α proteins （P<0.01）. ConclusionCPU Ⅱ has an obvious inhibitory effect on the growth of transplanted hepatocellular carcinoma Hep3B cells in nude mice. The mechanism of action is related to enhancing the expression of transcription factors HNF-4α and C/EBPα， thereby promoting the expression of miR-122 and inhibiting the expression of its target gene IGF-1R.
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Objective:To establish a method of measuring the contents of gallic acid, brevifolin, corilagin, geraniin, ellagic acid and rutin in Phyllanthus urinaria L. simultaneously with fingerprint study for analysis. Methods:Phyllanthus urinaria L. was extracted by ultrasound with 50% methanol. Chromatographic separation was performed on a Phenonmenex Luna C18 (4.6 mm×250 mm, 5 μm). The mobile phase consisted of acetonitrile (A) and 0.1% phosphoric acid aqueous solution (B) with gradient elution. The flow rate was 1.0 ml/min. The column temperature was 25 ℃, and the injection volume was 10 μl. The detection wavelength was 270 nm. HPLC fingerprints of Phyllanthus urinaria L. from different habitats was established. PCA and OPLS-DA were used to analyze the differences in chemical components of different habitats. Results:Gallic acid, brevifolin, corilagin, geraniin, ellagic acid and rutin showed good linearity at 0.042 8-0.641 6, 0.033 4-0.501 4, 0.142 2-2.133 1, 0.383 1-5.746 5, 0.063 1-0.946 2 and 0.019 2-0.287 8 μg, respectively. The average recovery rate of them was 103.65%, 96.39%, 101.85%, 95.04%, 98.79% and 98.33%, respectively. The HPLC fingerprints of different habitats contained 14 characteristic common peaks, and six compounds characteristic peaks were identified. PCA analysis showed that the chemical components of Phyllanthus urinaria L. from different habitats were different. Geraniin, ellagic acid and corilagin were screened by OPLS-DA. Conclusions:The method is efficient, accurate and sensitive, which can be used to measure the six components in Phyllanthus urinaria L.. The established HPLC fingerprint of different habitats combined with the measrurement method of six components can be used for the quality control and evaluation of Phyllanthus urinaria L..
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Objective:To explore the anti-hepatoma effect of compound <italic>Phylanthus urinaria</italic> Ⅱ （ CPU Ⅱ） by inhibiting the expression of the long non-coding RNA （lncRNA） colon cancer associated transcript-1 （CCAT1） and restoring the expression of microRNA let-7a. Method:Real-time fluorescence quantitative polymerase chain reaction （PCR） was used to detect the expression of lncRNA CCAT1 in normal liver cells （LO2 cells） and hepatocellular carcinoma HepG2 cells， and the differences in expression between these two types of cells were compared. The methylthiazolyl tetrazolium（MTT） assay was used to detect the proliferation of HepG2 cells after treatment with different concentrations of CPU Ⅱ and 5-fluorouracil（5-FU） for 24， 48 and 72 h. Hepatocellular carcinoma HepG2 cells were cultured <italic>in vitro </italic>and set into three gropes： cell control group， CPU Ⅱ low-dose group （0.8 g·L<sup>-1</sup>） and high-dose group （1.6 g·L<sup>-1</sup>）. Real-time PCR was used to detect the mRNA expression of lncRNA CCAT1， microRNA let-7a and its target genes high mobility group protein A2（HMGA2）， and N-RAS in each grope. Western blot was used to detect the protein expression of HMGA2， and Cyclin D<sub>1</sub> in each grope. Result:As compared with LO2 cells， expression of lncRNA CCAT1 in HepG2 cells was significantly up-regulated （<italic>P</italic><0.05）. Results of MTT assay showed that the 50% inhibiting concentration（IC<sub>50</sub>）<sub> </sub>of CPU Ⅱ and 5-FU on hepatocellular carcinoma HepG2 cells was 1.649， 0.044 648 g·L<sup>-1 </sup>respectively. As compared with the control group， CPU Ⅱ high-and low-dose groups （1.6， 0.8 g·L<sup>-1</sup>） significantly inhibited the proliferation of HepG2 cells （<italic>P</italic><0.05）， and the effect was most remarkable in CPU Ⅱ high-dose group （<italic>P</italic><0.05）. The results of Real-time PCR showed that as compared with control group， the expression of lncRNA CCAT1 mRNA was significantly inhibited in CPU Ⅱ high-and low-dose groups （<italic>P</italic><0.05）， and the expression of microRNA let-7a mRNA was obviously up-regulated in high-dose group （<italic>P</italic><0.05）， but the expression of HMGA2 mRNA in CPU Ⅱ high-and low-dose groups as well as the expression of N-RAS mRNA in CPU Ⅱ low-dose group were down-regulated （<italic>P</italic><0.05）. Western blot results showed that as compared with the cell control group， the protein expression of HMGA2 and Cyclin D<sub>1</sub> in CPU Ⅱ high-and low-dose groups （1.6， 0.8 g·L<sup>-1</sup>） was significantly down-regulated （<italic>P</italic><0.05）. Conclusion:CPU Ⅱ can inhibit the expression of lncRNA CCAT1， recover the expression of microRNA let-7a， and suppress the mRNA and protein expression of related downstream target genes in hepatoma cells line HepG2， thereby inhibiting the proliferation of hepatocellular carcinoma cells and exerting anti-hepatocellular carcinoma effect.
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Objective:To observe the effect and investigate the mechanism of compound Phyllanthus urinaria Ⅱ(CPU Ⅱ)on proliferation, apoptosis and autophagy of human hepatoma cell line Huh7. Method:Huh7 cells were cultured in vitro and divided into blank control group, high-dose CPU Ⅱ group (40 g·L-1),low-dose CPU Ⅱ group (20 g·L-1), and 5-FU group (0.04 g·L-1). Methye thiazolye telrazlium(MTT) assay was used to detect the inhibitory effect of CPU Ⅱ on proliferation of human hepatoma Huh7 cells. The apoptosis rate was observed by Annexin V-FITC/PI flow cytometry; the changes of autophagosomes in each group were observed by monodansylcadaverin (MDC) staining; Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression of phosphatidylinositol 3-kinase (PI3K), Serine-threonine protein kinase 2(Akt2), B-cell lympoma-2(Bcl-2) and Microtubule-associated protein 1 light chian 3(LC3Ⅱ) and Western blot was used to detect the protein expression of Akt2 and LC3Ⅱ. Result:CPU Ⅱ(40, 20 g·L-1) significantly inhibited the hepatoma cell line Huh7 and induced apoptosis, with an apoptosis rate of 51.72% and 19.74% respectively, significantly higher than that of control group (PPPPConclusion:CPUⅡ had obvious inhibitory effect on the proliferation of hepatoma cell line Huh7, and the mechanism may be related to inhibiting the activation of PI3K/Akt signaling pathway to induce apoptosis and autophagy.
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Aim To explore the anticomplementary ac-tivity and active constituents of Phyllanthus urinaria. Methods Being guided by bioactive screening,the anticomplementary constituents of P.urinaria were iso-lated and purified by solution partition and chromato-graphic techniques,and their structures were identified by 13C-NMR spectrum and the comparison of reported data. The anticomplementary activity and possible mechanism were assayed.Results The EtOAc fraction of the methanol extract of P.urinaria was shown to be the active fraction,and two compounds were purified from the fraction and were identified as corilagin and ellagic acid.The EtOAc and the two compounds signifi-cantly inhibited the hemolysis of the complement classi-cal pathway,with IC50values of 53.77 mg·L-1, 176.54 mg·L-1and 102.23 mg·L-1,respectively. While they just showed slight effect on inhibiting the hemolysis of the complement alternative pathway. All of them affected the formation of C3 convertase of the classical pathway. Conclusions The polyphenols are main anticomplentary constituents of P. urinaria,and its mechanism is related to inhibiting formation of C3 convertase of the classical pathway.
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Objective To study the chemical constituents of whole plant of Phyllanthus urinaria and their biological activity. Methods The chemical constituents were isolated and purified by repeated column chromatography over silica gel, Rp-C18 (reverse phase), MCI, and Sephadex LH-20, as well as semi-preparative HPLC. NMR spectroscopic analyses were used for the structure identification. In this case, the inhibitory rate of NO production of the isolated triterpenoids was evaluated. Results Seven triterpenoids, identified as 28-norlup-20(29)-ene-3,17β-diol (1), betulin (2), β-betulinic acid (3), 3-oxofriedelan-28-oic acid (4), oleanolic acid (5), 3R-E-coumaroyltaraxerol (6), and 3R-Z-coumaroyltaraxerol (7), were isolated and identified from the whole plants of P. urinaria. Compounds 1–5 exerted inhibitory effects against NO production in LPS-induced RAW 264.7 mouse macrophages with the inhibitory rate of NO production ranging from 4.0% to 27.8% at the concentration of 25 μmol/L. Conclusion To the best of our knowledge, this is the first report of compounds 1–4, 6, and 7 from the family Euphorbiaceae. Compounds 1–5 exhibited inhibitory effects against NO production in LPS-induced RAW 264.7 mouse macrophages.
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Objective To study the chemical constituents of whole plant of Phyllanthus urinaria and their biological activity. Methods The chemical constituents were isolated and purified by repeated column chromatography over silica gel, Rp-C18 (reverse phase), MCI, and Sephadex LH-20, as well as semi-preparative HPLC. NMR spectroscopic analyses were used for the structure identification. In this case, the inhibitory rate of NO production of the isolated triterpenoids was evaluated. Results Seven triterpenoids, identified as 28-norlup-20(29)-ene-3,17β-diol (1), betulin (2), β-betulinic acid (3), 3-oxofriedelan-28-oic acid (4), oleanolic acid (5), 3R-E-coumaroyltaraxerol (6), and 3R-Z-coumaroyltaraxerol (7), were isolated and identified from the whole plants of P. urinaria. Compounds 1–5 exerted inhibitory effects against NO production in LPS-induced RAW 264.7 mouse macrophages with the inhibitory rate of NO production ranging from 4.0% to 27.8% at the concentration of 25 μmol/L. Conclusion To the best of our knowledge, this is the first report of compounds 1–4, 6, and 7 from the family Euphorbiaceae. Compounds 1–5 exhibited inhibitory effects against NO production in LPS-induced RAW 264.7 mouse macrophages.
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Phyllanthus Urinaria L. (PUL) is a traditional Chinese medicine used to treat hepatic and renal disorders. However, the mechanism of its hepatoprotective action is not fully understood. In the present study, blood biochemical indexes and liver histopathological changes were used to estimate the extent of hepatic injury. GC/MS and LC/MS-based untargeted metabolomics were used in combination to characterize the potential biomarkers associated with the protective activity of PUL against CCl-induced liver injury in rats. PUL treatment could reverse the increase in ALT, AST and ALP induced by CCl and attenuate the pathological changes in rat liver. Significant changes in liver metabolic profiling were observed in PUL-treated group compared with liver injury model group. Seventeen biomarkers related to the hepatoprotective effects of PUL against CCl-induced liver injury were screened out using nonparametric test and Pearson's correlation analysis (OPLS-DA). The results suggested that the potential hepatoprotective effects of PUL in attenuating CCl-induced hepatotoxicity could be partially attributed to regulating L-carnitine, taurocholic acid, and amino acids metabolism, which may become promising targets for treatment of liver toxicity. In conclusion, this study provides new insights into the mechanism of the hepatoprotection of Phyllanthus Urinaria.
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Animaux , Humains , Mâle , Rats , Acides aminés , Métabolisme , Tétrachloro-méthane , Lésions hépatiques dues aux substances , Métabolisme , Médicaments issus de plantes chinoises , Chromatographie gazeuse-spectrométrie de masse , Foie , Métabolisme , Métabolomique , Phyllanthus , Chimie , Rat Sprague-Dawley , Acide taurocholique , MétabolismeRésumé
Objective: To study the chemical constituents of Phyllanthus urinaria. Methods: Compounds were isolated and purified by the normal phase silica gel, Sephadex LH-20, MCI gel, RP-18, and semi-manufactured preparation HPLC method. Their structures were identified by the methods of 1H-NMR and 13C-NMR combined with physicochemical property. Results: Sixteen compounds were isolated from the petroleum ether and ethyl acetate fraction in 95% ethanol extract of P. urinaria and their structures were gallic acid (1), caffeic acid (2), ethyl gallate (3), methyl gallate (4), 4-ethoxybenzoic acid (5), diisobutyl phthalate (6), dibutyl phthalate (7), (4R,6R)-2,3-dihydromenisdaurilide (8), (4R,6S)-2,3-dihydromenisdaurilide (9), aquilegiolide (10), menisdaurilide (11), cassipourol (12), (9Z,12Z)-nonadeca-9,12-dienoic acid (13), methyl linoleate (14), stigmasterol (15), and (8R,8'S,7S)-4'-(3″- methoxyrhamnopyranosyl) oxy-8'-hydroxy-3,3',4-trimethoxy-8-hydroxymethyl-lign-7-9'-lactone (16). Conclusion: Compounds 5-7, 10, 11, 13, and 14 are isolated from this plant for the first time, and compounds 8, 9, 12, and 16 are first isolated from the plants in genus Phyllanthus L. for the first time.
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This study was aimed to establish the HPLC fingerprint ofPhyllanthus urinaria Linn and Phyllanthusamarus Linn, in order to provide evidences for the study on material basis. Analysis was performed on an INDUSTRIES Epic C18 120A (5μm, 250 mm × 4.6 mm) column eluted with the acetonitrile (A) - water (0.1% phosphoric acid, V/V) gradient system as mobile phase. The wavelength was 254 nm and the flow rate was 1mL·min-1. The column temperature was 30℃. The injection volume was 10 μL. The results showed that the HPLC fingerprint ofPhyllanthus urinaria L. andPhyllanthus amarus L. were established. It was concluded that the method was simple, accurate and reproducible. This study provids experimental data for rapid quality identification and comprehensive evaluation ofPhyllanthus urinaria L. andPhyllanthus amarus L..
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Objective: To establish the HPLC fingerprint and its evaluation system for the whole plant of Phyllanthus urinaria. Methods: Diamonsil C18 column (250 mm × 4.6 mm, 5 μm)was used with acetonitrile and 0.2% acetic acid solution in gradient elution mode. The detective wavelength was 270 nm and the flow rate was 1.0 mL/min. The fingerprints for the whole plant of P. urinaria and phenolic acids part from nine different sources were set up, the similarity assay for the whole plant of P. urinaria from nine different sources was carried out to evaluate their quality by Chinese material medica (CMM) fingerprint similarity evaluation system ( 2004A edition), and the peaks were identified by comparing the reference substance with LC-MS/MS. Results: The common mode of HPLC fingerprints for the whole plant of P. urinaria were set up. There were 16 common peaks in the fingerprints and eight peaks were identified. The cluster analysis and principal component analysis were applied to studying the HPLC fingerprint and chemical pattern recognition. Conclusion: The combination of fingerprint and chemical pattern recognition is an effective method for the quality control of the whole plant of P. urinaria. The study lays the foundation for the full quality evaluation for the whole plant of P. urinaria.
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<p><b>OBJECTIVE</b>To explore the effects of the extract from Phyllanthus urinaria L. on hepatitis B virus (HBV) replication and expression in HBV transient transfection model in vitro.</p><p><b>METHODS</b>The eukaryotic expression plasmid pHBV1.1, which contains 1.1-fold-overlength genome of HBV, was transfected into the human hepatoma cell line, HepG2, to establish and assess the HBV transient transfection model. The extract from Phyllanthus urinaria L. was prepared in different concentrations and methyl thiazolyl tetrazolium was used to detect the maximum nontoxic concentration of the drug. The extract from Phyllanthus urinaria L. were added into the transfected cell, at the concentrations of 0.8, 0.2 and 0.05 g/L, respectively. Four days after drug application, enzyme-linked immuno sorbent assay was used to detect the concentration of HBsAg in the supernatants, Southern blot was applied to analyze HBV DNA level, and Western blot was used to detect the expression of HBcAg in cells.</p><p><b>RESULTS</b>After the transfection of plasmid pHBV1.1 into HepG2 cells, the concentration of HBsAg in supernatants was increased obviously as compared with that of the normal cells (P<0.05), and all expected HBV replicative intermediates were confirmed by Southern blot analysis, which ensured the successful establishment of the HBV transient transfection model. After the application of drugs at the concentrations of 0.8 and 0.2 g/L, the level of HBsAg was obviously decreased in the supernatants, as compared with that of the virus group (P<0.05); Southern blot showed that the level of HBV rc DNA, ds DNA, ss DNA was obviously reduced compared with that of the virus group (P<0.01); Western blot revealed that the expression of HBcAg in the drug group was obviously inhibited, as compared with that of the virus group (P<0.01).</p><p><b>CONCLUSIONS</b>The extract from Phyllanthus urinaria L. obviously inhibited replication and expression of HBV in HBV transfected cell lines in vitro, thus exerting distinctive anti-HBV effects.</p>
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Humains , Cellules HepG2 , Hépatite B , Traitement médicamenteux , Virus de l'hépatite B , Physiologie , Phyllanthus , Extraits de plantes , Pharmacologie , Transfection , Réplication viraleRésumé
<p><b>OBJECTIVE</b>To observe the change in the number of antibodies of preneoplastic hepatocellular carcinoma (HCC) using early treatment by Compound Phyllanthus Urinaria L. (CPUL) on patients with preneoplastic hepatitis B virus (HBV)-associated HCC.</p><p><b>METHODS</b>A total of 102 cirrhosis patients with regenerative or dysplastic nodules whose sera were tested positive for at least one of these six proteins (five up-regulated genes URG4, URG7, URG11, URG12 and URG19, and one down-regulated gene DRG2) were assigned randomly to two groups using continual random codes by SPSS software. Fifty-two patients were in the treatment group and 50 patients were in the control group. CPUL was used in the treatment group for 3 years, while the control group did not receive any treatment. The changes in HBV-DNA level, number of antibodies, and hepatocarcinogenesis occurred were observed. Patients who did not develop HCC were followed up for another 2 years.</p><p><b>RESULTS</b>HBV-DNA levels decreased ⩾2log in 22.2% (10/45) of patients in the treatment group in contrast to only 5.0% (2/40) of patients in the control group (P=0.0228). The number of antibodies that were tested positive in the treatment group (1.08±1.01) was significantly lower compared with the control group (2.11±1.12) after 24 months of drug treatment (P<0.01). Both the positive rates of anti-URG11 (33/52) and anti-URG19 (31/52) were over 60% at baseline in the two groups, and were decreased to 48.1% (25/52) and 46.2% (24/52) respectively at 36 months of drug treatment, while the rates increased to 68.0% (34/50) and 66.0% (33/50) respectively (P=0.0417, P=0.0436) in the control group. The positive rate of anti-DRG2 was increased to 55.8% (29/52) at 36 months of drug treatment, while in the control group was decreased to 36.0% (18/50, P=0.0452). Among the 102 patients who developed HCC, 2 were in the treatment group and 9 were in the control group, meaning that a significant difference between the two groups (P=0.0212). In 11 patients who developed HCC, anti-URG11 and anti-URG19 were always positive, while anti-DRG2 was negative. Patients newly developing HCC were 6 (20.0%) in the control group, and only one (2.5%) in the treatment group (P=0.0441) during 2-year follow-up after the end of the treatment.</p><p><b>CONCLUSIONS</b>Anti-URG11, anti-URG19 and anti-DRG2 could be used as early markers in the prediction of the therapeutic efficacy of CPUL in treating preneoplastic HCC. CPUL is useful in preventing or delaying the development of HBV-associated cirrhosis to HCC.</p>
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Humains , Anticorps antiviraux , Sang , Carcinome hépatocellulaire , Thérapeutique , Virologie , ADN viral , Cellules HepG2 , Virus de l'hépatite B , Génétique , Allergie et immunologie , Virulence , Tumeurs du foie , Thérapeutique , Virologie , Phyllanthus , Chimie , Extraits de plantes , Utilisations thérapeutiques , États précancéreux , VirologieRésumé
OBJECTIVE: To optimize the extraction technology of thrombolytic components predominantly as corilagin from Phyllanthus urinaria.METHODS: The extraction technology was optimized by orthogonal experiment with solvent amount,extracting time and extracting times as factors,and with the comprehensive score of the yield of extract and corilagin content as indexe.The impurity-removing efficacy by centrifugation,acid/base method or ethanol precipitation was evaluated.RESULTS: The optimal extraction technology was as follows: using 8-fold volume of 30% ethanol as solvent for reflux extraction of 3 times(2 hours/ time).The best impurity-removing efficacy was achieved when 90% ethanol was used for precipitation.CONCLUSION: The extraction technology is simple and stable with high yield of active components,and it provides theoretic basis for the industrial production.
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Objective To evaluate the protective action of Phyllanthus orinaria(PU)for immune liver injury in mice.Methods Forty-eight NIH mice were allocated randomly to high-dose(HG)PU group,low-dose(LG)PU group,bifenbate group(BG),normal control group(NCG),and model group(MG).From the first day of establishing the models,20 g ? kg-1 of PU solution was given for gastric perfusion(GP)to HG PU group,10 g ? kg-1 of PU solution for GP to LG PU group,0.15 g?kg-1 of bifenbate solution for GP to BG group,and the equal amount of normal saline to the normal control group and the model group,qd for 12 days.Serum ALT level and visceral parameters of liver and spleen were determined,and the liver pathological feature was also examined.Results The ALT level,parameters of liver and spleen were markedly decreased in BG group(P 0.05).Conclusion PU has a good effect on counteracting the immune live injury in mice by markedly decreasing the activity of serum transaminase and visceral parameters of liver and spleen,and improving the necrosis of hepatic cells.
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Objective To evaluate the in-vivo effect of Compound Phyllanthus urinaria Ⅱ(CPU Ⅱ) on duck hepatitis B virus(DHBV).Methods Thirty ducks with congenital infection of DHBV,which were DHBV-DNA positive confirmed by polymerase chain reaction(PCR),were randomly divided into 5 groups: DHBV control group,high-,middle-,and low-dosage CPU Ⅱgroups(in the dose of 33.6,18.6 and 8.4 g-1?d-1,respectively),and Lamivudine group(20 mg?kg-1?d-1).The serum DHBV-DNA level of all ducks was detected by quantity real-time fluorescence PCR before and after medication.Results On the 7th,14th,21st,and 28th day of medication and on the 5th day of suspension of medication,the serum DHBV-DNA level in both of the high-and middle-dosage CPU Ⅱ groups was obviously lower than that before medication and than that in the DHBV control group(P
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Objective To establish culture method for Phyllanthus urinaria. Methods To study the possible effective factors of culture condition by comparing with different explants,sucrose,plant growth substance,and its ratio. Results The inductivity of stem was the highest about 55.56%,but callus of leaves could not be induced.6-BA was the most important factor on callus induction,followed by 2,4-D NAA and sucrose. Conclusion The optimal medium to induce and subculture is MS+2,4-D 0.5 mg/L+6-BA 1.0 mg/L+NAA 0.1 mg/L+sucrose 10 g/L.
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Objective To investgate the inhibitory effect of Phyllanthus Urinaria L compound on proliferation of hepatoma cell HePG2 in vitro and to explore its mechanism.Methods The influence of different concentrations of Phyllanthus Urinaria L compound at different time on HePG2 proliferation was compared by MTT colorimetric assay and cell growth curve assay.The cell apoptotic rate and morphological changes of HePG2 were observed by flow cytometry,fluorescence microscope and electron microscope.Results Phyllanthus Urinaria L compound can inhibit the proliferation of human hepatoma cell line HePG2.Within a certain limit of concentrations,the higher the concentration and the longer the acting time,the stronger the inhibition.Co-cultured with 500 ? g/mL Phyllanthus Urinaria L compound for 72 h,the inhibitory rate reached 93.58 % and IC50 was 240 ? g/mL.Phyllanthus Urinaria L compound at different concentrations had an certain effect on inducing cell apoptosis.Conclusion Phyllanthus Urinaria L compound can inhibit the proliferation of hepatoma cell,and its mechanism may be related with the induction of hepatoma cell HePG2 apoptosis.