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Objective:To investigate the intervention of <italic>Hedyotis diffusa</italic> （HDW） on colitis associated cancer （CAC） model mice and explore its mechanism. Method:The CAC mouse model was established by synergistic action of azoxymethane （AOM） and dextran sulfate sodium （DSS）. The intervention of HDW on CAC mice was evaluated by disease activity index （DAI）， colonic tissue morphology， pathological injury score and tumorigenesis rate. Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF-MS） and multivariate statistical analysis were used to analyze the metabonomics of mice serum and to explore the mechanism of HDW intervention on CAC. Result:HDW could significantly improve the general condition of CAC mice， decrease DAI， colon gross morphological score， histopathological score and tumorigenesis rate. Compared with the normal group， 38 kinds of differential metabolites were screened in the model group， including 11 potential biomarkers， involving 11 main metabolic pathways. HDW could significantly regulate 9 kinds of differential metabolites ［niacinamide， uridine， 4-pyridoxic acid， LysoPC （18∶0）， LysoPE （0∶0/20∶0）， myo-inositol， purine， sphinganine 1-phosphate and tetradecanedioic acid］ in the model group， including 2 kinds of potential biomarkers （myo-inositol and niacinamide）， and HDW could regulate nicotinate and nicotinamide metabolism and inositol phosphate metabolism. Conclusion:HDW has a therapeutic effect on CAC， which may be achieved by regulation of energy metabolism and glucose metabolism.
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To identify the composition of iridoids from Hedyotis diffusa Willd and explore the mechanism on its anti-renal fibrosis effect based on network pharmacology, LC-Q/TOF-MS (liquid chromatograpy-quadrupole/time of flight mass spectrometry) was used to analyze the iridoid ingredients and the related targets of renal fibrosis were obtained by DisGeNET database and MalaCards database. The potential targets were screened by SYBYL-X7.3 software. We then imported the identified ingredients and potential target genes into Cytoscape3.7.1 to construct the compound-target network and the protein-protein interaction (PPI) network. Finally, the gene ontology (GO) functional enrichment analysis and KEGG pathway enrichment analysis of the selected core genes were made to explore the mechanism of iridoids against renal fibrosis. There were 10 active iridoid compounds and 111 corresponding targets including dimethylarginine dimethylaminohydrolase 1 (DDAH1), heparanase (HPSE), human kirsten rat sarcoma viral oncogene (KRAS), moesin (MSN), etc. in compound-target network. The GO functional enrichment analysis obtained 211 GO entries. Twenty related signal pathways including Toll-like receptor signaling pathway, transforming growth factor-beta (TGF-β) signaling pathway, renal cell carcinoma signaling pathway, and the Janus kinase/signal transducer and activator of tran-ions (Jak-STAT) signaling pathway were selected by KEGG enrichment analysis. We preliminarily investigated the mechanism of the iridoid compounds on renal fibrosis to provide guide information for the subsequent experimental research and clinical application.
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@#This research used inter simple sequence repeat(ISSR)markers to analyze the genetic diversity of Hedyotis diffusa Willd. from different origins. A total of 23 samples of Hedyotis diffusa Willd. in the Guangxi Zhuang Autonomous Region, Guangdong, Hunan, Zhejiang, Fujian, Jiangxi and Anhui, respectively were collected. 150 ISSR primers were used to amplify PCR and then POPGENE1. 32, NTSYS2. 10 software were used to analyze genetic diversity. 11 primers were screened, 115 polymorphic bands were amplified, the polymorphism ratio was 85. 22%, the number of alleles(Na)was 1. 852 2, the effective allele(Ne)was 1. 543 4, Neis gene diversity index(H)was 0. 316 5 and Shannon′s information index(I)was 0. 470 0. The results of cluster analysis show that the Hedyotis diffusa can be divided into three clades. The conclusion is that ISSR molecular markers can provide a insight for the identification of Hedyotis diffusa Willd. .
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Objective: To investigate the effects of Hedyotis diffusa Willd (HDW) on the proliferation, cell cycle, apoptosis, migration and invasion of human renal carcinoma cells, and to explore the possible mechanisms. Methods: After human renal adenocarcinoma ACHN cells and human renal proximal tubule HK-2 cells were treated with HDW for 24 h, the cell proliferation was detected by CCK-8 assay, and the cell cycle and apoptosis were detected by FCM assay. The phenotypes of ACHN and HK-2 cells treated with HDW were observed by light microscopy and rhodamine staining. The expressions of mesenchymal-epithelial transition (MET)-related proteins were detected by immunofluorescence staining and Western blotting, respectively. The effects of HDW on the migration and invasion of ACHN and HK-2 cells were detected by wound-healing and Transwell chamber assay. RNAsequencing was used to detect the effect of HDW on the transcriptome in ACHN cells. The expression levels of RAP1 pathway-related genes [RAP1 GTPase activating protein (RAP1GAP), RAS guanyl releasing protein 2 (RASGRP2), RAP guanine nucleotide exchange factor 3 (RAPGEF3), membrane-associated guanylate kinase, WW and PDZ domain containing 1 (MAGI1) and G protein subunit alpha i1 (GNAI1)] were detected by real-time fluorescent quantitative PCR. The expression levels of mitogen-activate protein kinase (MAPK) pathway-related proteins [c-Jun N -terminal kinase (JNK), phospho-JNK (p-JNK), protein kinase B (PKB, Akt), phospho-Akt (p-Akt), extracellular signalregulated kinase (ERK) and phospho-ERK (p-ERK)] were detected by Western blotting. Results: HDW selectively inhibited the proliferation of ACHN cells (P < 0.01), blocked cell cycle at S-phase (P < 0.01), and induced apoptosis (P < 0.01). After treatment with HDW, the morphology of ACHN cells significantly changed. HDW promoted the expressions of MET-related proteins (E-cadherin 1 and β-catenin) in ACHN cells, and inhibited the expressions of epithelial-mesenchymal transition (EMT)-related proteins (Vimentin and Snail1) (all P < 0.01). HDW inhibited the migration and invasion of ACHN and HK-2 cells (all P < 0.01), and there was no significant difference between ACHN and HK-2 cells. HDW affected the expressions of cell cycle-related genes and transcription factor E2F and Myc target genes, and activated the p53 signaling pathway. HDW significantly downregulated the expression levels of RAP1GAP, RASGRP2, RAPGEF3, MAGI1 and GNAI1 (all P < 0.000 1) and the phosphorylation level of JNK in ACHN cells. Conclusion: HDW may selectively inhibit cell proliferation, and promote MET, cell cycle arrest and apoptosis of ACHN cells by inhibiting RAP1-JNK signaling pathway.
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OBJECTIVE@#To investigate the therapeutic effects of Hedyotis diffusa Willd.on type Ⅱ collagen-induced rheumatoid arthritis in rats.@*METHODS@#According to the random number table, 60 SD rats were divided into the normal control group (=10, normal saline) and model group (=50).The collagen-induced arthritis model was established with the injection of type Ⅱ collagen into the back in rats other than the normal group and evaluated by arthritis score, then the model rats were randomly divided into model group (normal saline), tripterygium wilfordii polyglycoside (GTW) 6 mg/kg group (daily dose:0.4 mg/kg), HD 3, 6, 12 g/kg groups (daily dose:3, 6 and 12 g/kg, respectively), with 10 rats in each group. The rats were treated with corresponding agents by intragastric administration.The arthritis index and the pain threshold of all rats at different time points were observed and measured weekly.After treated by intragastric administration for 28 days, all rats were killed to measure the changes of serum cytokine levels including interleukin 1β (IL-lβ), tumor necrosis factor a (TNF-a), prostaglandin (PGE), receptor activator for nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG).@*RESULTS@#Compared with the control group, the arthritis index and the serum levels of IL-lβ, TNF-a, PGE, RANKL, OPG and RANKL/OPG of the model group were increased significantly (<0.05), the pain threshold of the model group was decreased significantly (<0.05); compared with the model group, the arthritis index and the serum levels of IL-lβ, TNF-a, PGE, RANKL, OPG and RANKL/OPG of the GTW group, HD low-dose, medium-dose, high dose groups were decreased significantly (<0.05), the pain threshold of the model group was increased significantly (<0.05).@*CONCLUSIONS@#Hedyotis diffusa Willd.can significantly reduce arthritis index and increase pain threshold, reduce the level of IL-lβ, TNF-a, PGE, RANKL, OPG, and RANKL/OPG, then can prevent CIA effectively.
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Animals , Rats , Arthritis, Experimental , Arthritis, Rheumatoid , Collagen Type II , Hedyotis , RANK Ligand , Rats, Sprague-DawleyABSTRACT
To explore the regulatory effect and relevant mechanisms of the fraction of Hedyotis diffusa and Scutellaria barbata herb couple(YDW11) on polarization of macrophage between M1/M2 phenotypes.RAW264.7 cells were induced with LPS/IFN- or IL-4/IL-13 to establish M1 or M2 macrophage cell model. MTT assay was used to measure the cell cytotoxicity of YDW11. Griess reaction was used to detect the changes of nitrite accumulation in the cell supernatant. Trans-well assay was used to measure the migration capability. QRT-PCR was used to assay mRNA expressions of iNOS, IL-1, Arg-1 and MR. Western blot was used to detect the effect of YDW11 on iNOS and Arg-1 protein expressions. Taqman MicroRNA RT-PCR was used to detect the effect of YDW11 on miR155 expression under M1 and M2 phenotype conditions. In addition, MS-UPLC assay was carried out to identify the constituents in YDW11. The results showed that the ethyl acetate of H. diffusa and S. barbata extracted in 1:1 ratio with water (YDW11) showed the activity in suppressing the nitrite content in M1 macrophages without cytotoxicity. YDW11 also inhibited the migration of breast cancer cells with the help of M2 macrophages by blocking their polarization towards M2. YDW11 decreased iNOS, IL-1, Arg-1and MR mRNA expressions and iNOS and Arg-1 protein expressions. YDW11 down-regulated miR155 expression in M1 phenotype, and up-regulated miR155 expression in M2 phenotype. Based on MS-UPLC,four compounds were identified in YDW11, including 4'-hydroxyacetophenone, scutellarin, luteolin and apigenin. YDW11 inhibited M1/M2 phenotypes of macrophages by regulating the expression of miR155.
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Objective:To optimize the extraction technology and evaluate antioxidant activity of total flavonoids from Hedyotis dif-fusa. Methods:The content of kaempferol was detected by an HPLC method,and spectrophotometry was used to detect the content of total flavonoids. Results: The optimum extraction conditions were investigated by orthogonal design with the extraction quantities of kaempferol and total flavonoids as the evaluation indices. The antioxidant activities of the extracts were detected based on the clearance of hydroxyl radical and DPPH·model. The optimum extraction conditions were as follows:10-fold amount of 80% methanol and with ultrasonic extraction for 15 min. The clearance rate of 1ml methanol extract for scavenging DPPH· and ·OH was 73.89% and 91.27%,respectively.Conclusion:The optimum technology is stable and feasible for the extraction of Hedyotis diffuse. The extract shows good in vitro antioxidant properties, which provides powerful reference for the future development of relevant antioxidant prod-ucts.
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Objective: Based on the tumor-bearing rat model, using rutin, quercitrin, and isoquercitrin from Hedyotis diffusa as the research object, the pharmacokinetics of those three flavonoid glycosides in the pathological state was studied. Methods: Establishing a method for the comparison of the pharmacokinetics of flavonoid extracts in normal rats and tumor-bearing rats, which was analyzed by HPLC-MS/MS in subcutaneous tumor models of SD rat made with tumour cell of Walker-256. Results: Compared with the pharmacokinetics parameters of flavonoid glycosides in normal rats, the Cmax and AUC0-∞ of rutin, quercetin, and isoquercitrin in the tumor-bearing rats were significantly decreased, t1/2z was prolonged, and the metabolic time of three components was prolonged to 24 h, which revealed the effect of pathological condition on the pharmacokinetic characteristics of flavonoid glycosides. Conclusion: The method established in this study is simple, fast, sensitive, and suitable for the pharmacokinetic study of flavonoid glycosides in rats in vivo. The pharmacokinetic characteristics of flavonoids in normal and tumor-bearing rats are different.
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Objective: To establish a method of rapid analysis for simultaneous determination of oleanolic acid and ursolic acid in Chinese materia medica (CMM) of Verbena officinalis, Ligustrum lucidum, Prunella vulgaris, Hedyotis diffusa, Patrinia scabiosaefolia, Eriobotrya japonica, Crataegus pinnatifida, Chaenomeles Fructus papaya. Methods: The analyses of preparation were conducted by accelerated solvent extraction (ASE), methanol was used as solvent extraction, and the static extraction time was 6 min. Separation was carried out on Acclaim C30 Thermo column with acetonitrile and 0.2% acetic acid (85:15) as mobile phase, flow rate was 0.3 mL/min, and UV detection wavelength was set at 205 nm. Results: After 10 min sample extraction time, oleanolic acid and ursolic acid reached baseline separation, and no sample matrix was interfered, the linear correlation coefficient was over 0.999, the average recovery between 95.8% and 102.7%; Compared to Chinese Pharmacopoeia 2015, the determination of average mass fraction difference was 3.7% and 5.8%. Conclusion: This method is simple, fast, and suitable for the analysis of these drugs, and it can be used for content determination of oleanolic acid and ursolic acid for eight kinds of CMM.
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Hedyotis diffusa and H. corymbosa belong to the genus Hedyotis Linn. in family Rubiaceae. They shared similar heat-clearing and toxin-resolving effects and pharmacological activities, such as anti-cancer activity, and H. diffusa is frequently adulterated or substituted by H. corymbosa in clinical application. But whether they can completely replace with each other or whether the therapeutic effect could be changed after the substitution, is still a problem needs to be resolved. Therefore, the two herbs were compared in terms of the textual research, medical history, mixed use, commercial medicine status, macro-characters, micro-characteristic, chemical compositions and pharmacologic activities, and found that they shared similar macro-characters and micro-characteristic but were obviously different in chemical composition and pharmacologic activities, and now they could be identified quickly and accurately by molecular identification method. Therefore, we suggest that they should be used as two different medicinal materials at this stage.
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OBJECTIVE: To explore the effect of total coumarins isolated from Hedyotis diffusa (total coumarins from Hedyotis diffusa, TCHD) on proliferation inhibition of leukemia cells, and to explore its related mechanism. METHODS: The purity of TCHD prepared by ethanol reflux extraction was tested by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) system. The cells (KG-1 Kasumi-1, THP 1 cells, U937 cells and K562 cells) were treated with TCHD(0.02, 0.04, 0.06, 0.08, 0.10 mg·mL-1) for 24 or 48 h, the inhibitive effect of TCHD on cells growth were determined by MTT method. After Kasumi-1 cells were incubated with TCHD for 24 h, the apoptosis of cells were analyzed by flow cytometry stained with Annexin V/PI. The expression levels of caspase-3, caspase-8, caspase-9, PARP and Bcl-2 family protein were assayed by Western blot. RESULTS: TCHD in certain concentration range could markedly inhibit the proliferation of AML cells, their IC50 on Kasumi-1, THP-1 KG-1, U937 and K562 cells were 0.077, 0.083, 0.096, 0.087, 0.096 mg·mL-1 for 24 h, and 0.059, 0.067, 0.072, 0.064, 0.068 mg·mL-1 for 48 h. TCHD has significant inhibitory effect on Kasumi-1, which was stronger than those on other cell lines, and showed a dose- and time-dependent manner(r=0.357, P<0.05). The apoptotic proportion of Kasumi-1cells in 0, 0.02, 0.04, 0.06, 0.08, 0.10 mg·mL-1 TCHD treatment groups for 24 h were (5.33±0.41)%, (7.99±0.45)%, (10.22±0.32)%, (20.10±1.99)%, (28.66±0.67)% and (33.24±2.12)%, respectively. After treated with TCHD(0.02-0.06 mg·mL-1) for 24 h, G0/G1 phase ratio of Kasumi-1 detected by flow cytometry were (51.43±3.21)%, (62.91±2.35)% and (76.42±4.14)%, respectively, which were significantly higher than that of the control group (35.8±5.25)% (P<0.05).Western blot results showed that different concentrations of TCHD could activate caspase-8, caspase-9, caspase-3 and PARP, promote the expression of cyto-C, down-regulate the cyclin E and CDK6, CDK2, p-CDK2 and cyclin D1 protein, and up-regulate the expression of p21 proteinin concentration- dependent manner(P<0.01). CONCLUSION: TCHD can obviously inhibit the proliferation of Kasumi-1 in a dose- and time-dependent manner, which may relate to the apoptosis of Kasumi 1 induced by activating caspase-3, 9, PARP protein through the mitochondrial pathways and Kasumi-1 cell block in G0/G1 phase through the influence of CDK2, p-CDK2, CDK4/6, cyclin E, cyclin D1 and p21.
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Objective To study and compare the metabolism of Hedyotis diffusa flavonoid extract in intestinal flora from human, normal rats and pseudo germ-free rats in vitro. Methods Intestinal flora of human, normal rats and pseudo germ-free rats were incubated with Hedyotis diffusa extract, rutosid, isoquercitrin and quercitrin under anaerobic conditions at 37℃.High-performance liquid chromatography with diode-array detection ( HPLC-DAD ) was used for the analysis of components and metabolic products.The trends of metabolic transformation of the various components were analyzed according to the changes in chromatographic peak areas at different incubation time points. Results The components of Hedyotis diffusa extract and its monomer were quickly metabolized by human and normal rat intestinal flora and the main metabolite was quercetin.However, they were hardly metabolized by intestinal flora from pseudo germ-free rats. The metabolism effect of intestinal flora was weaker on components than on the corresponding monomers. Conclusion By comparing flavonoids metabolism in intestinal flora from human, normal rats and pseudo germ-free rats, the important role of intestinal microflora in the metabolism of flavonoids is further confirmed.
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Objective To explore an UPLC method for simultaneous content determination of the five nucleosides (cytidine, uridine, adenine, thymidine and adenosine) in Hedyotis diffusa and its adulterants; To compare the content differences.Methods The analysis was performed on a BEH C18 column (2.1 mm×50 mm, 1.7 μm) by UPLC eluted with acetonitrile and water in gradient mode. The flow rate was 0.5 mL/min; the detection wavelength was set at 254 nm; the column temperature was set at 30℃.Results Five nucleosides have good linear relationship, precision, stability, and repeatability according to the requirements of the methodology determination. The recoveries were among 98.7%–101.5%. Five nucleosides in Hedyotis diffusa and its adulterants from different areas were determined by the UPLC method.Conclusion The method is certified to be simple, rapid, accurate and reliable, which can be used for the determination of nucleosides in Hedyotis diffusa and its adulterants.
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Objective To establish a method for quality control of oleanolic acid and ursolic acid in Qingdu tablet. Methods The content of oleanolie acid and ursolic acid in Qingdu tablet were determined by high performance liquid chroma-tography(HPLC). The analytical column was kromasil-C18 (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of methanol-0.5% phosphoric acid solution (88:12). The UV detection wavelenghth was 210 nm. The flow rate was 1.0 ml/min. Results Oleanolic acid and ursolic acid showed a good linearity in range of 0.06-0.54μg (R=0.999 3) and 0.30-2.70μg (R=0.999 7). The average recovery rates of Oleanolic acid and ursolic acid were 100.48% and 102.21%, respectively. Conclusions This method is simple, rapid and accurate. It could be used for the quality control and quality study of the Qingdu tablet.
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Objective: To investigate the intestinal absorption characteristics of five kinds of flavonoids from Hedyotis diffusa extract in rats. Methods: The in situ rat single-pass intestinal perfusion model was used. The contents of rutin, isoquercitrin, quercetin, kaempferol, and quercetin in perfusates were determined by HPLC-DAD, and the rat intestinal absorption parameters of flavonoids were calculated. Results: In perfusion model, the effective permeability coefficients (Peff) of all components were low. The absorption rate constant (Ka) and Peff had no significant difference when the concentration of total flavonoids from H. diffusa was 0.5-4.0 g/L. In the 2.0 g/L concentration, Ka of rutin, isoquercitrin, quercetin, kaempferol, and quercetin were 0.0113, 0.0154, 0.0102, 0.0305, and 0.0275 min-1, respectively; The Ka sequences of rutin and isoquercitrin in different intestinal segments were ileum > duodenum > jejunum ≈ colon and jejunum > duodenum > ileum ≈ colon. Conclusion: The absorption of the five flavonoids from H. diffusae is a first-order process with the passive diffusion mechanism. The absorption rates of each flavonoid are significantly different. The absorption rate of flavonoid glycosides is lower than that of aglycones. The flavonoids from H. diffusa could be absorbed in all the intestinal segments. The best parts of intestine to absorb rutin and isoquercitrin are ileum and jejunum.
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Hedyotis diffusa is an antioxidant, antibacterial Chinese herbal medicine which has anti-tumor, antioxidant, antibacterial, enhance the effect of nonspecific immunity and protection of the nervous system. Clinical application shows thatHedyotis diffusa has good efficacy on treatment of malignant tumors and inflammatory diseases. Referred to some papers published at home and abroad, this paper summarized from the aspects of active ingredient and antitumor effect. Results showes that its anti-tumor effect exactly, anti-tumor mechanism may be associated with a variety of molecular mechanisms, which remains to be further in-depth study.
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OBJECTIVE: To investigate whether the ethanol extract of Oldenlandia diffusa (EEHD) have impact on the TGF-β1-induced epithelial mesenchymal transformation (EMT) of human lung adenocarcinoma cell H358 in vitro. METHODS: The cell model to study EMT of H358 cells was established with 5 ng·mL-1 TGF-β1 for 24 h. Then, after being mixed with low, medium and high concentration of EEHD respectively for 48 h, H358 cells were collected for detection of protein and mRNA expression of epithelial marker E-cadherin and mesenchymal cell marker vimentinby Western-Blot and quantitative PCR. RESULTS: The concentration of 5 ng·mL-1 TGF-β1 can successfully induce the transformation from epithelialphenotype to mesenchymal phenotype. The concentration of EEHD is positively correlated with increasing expression of E-cadherin, while negatively correlation with the decreasing expression of vimentin. CONCLUSION: The EEHD have partially reversal effect of epithelial mesenchymal transformation on lung adenocarcinoma cell induced by TGF-β1.
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OBJECTIVE: To develop an HPLC method for simultaneous determination of multiple-components in Hedyotis diffusa Willd. METHODS: The HPLC analysis was carried out on a C18 column (4.6 mm×250 mm, 5 μm) by gradient elution with acetoni-trile-water[both containing 0.1‰ (V/V) acetic acid] as mobile phase at a flow rate of 0.8 mL·min-1, the column temperature at 35°C, and the detection wavelength was set at 238 nm. External standard method and quantitative analysis of multi-components by single marker (QAMS) method were adopted for simultaneous determination of six components in Hedyotis diffusa Willd, respectively. RESULTS: The linear ranges for asperulosidic acid, quercetin-3-O-[2-O-(6-O-E-feruloyl)-β -D-glucopyranosyl]-β-D-glucopyrano-side, kaempferol-3-O-[2-O-(6-O-E-feruloyl)-β-Z) -gfucopyranosyl]-β-D-galactopyranoside, (E)-6-O-p-coumaroyl scandoside methyl ester, (E)-6-O-feruloyl scandoside methyl ester, (Z)-6-O-p-coumaroyl scandoside methyl ester were 2.34-93.50, 2.61-104.33, 0.67-26.69, 3.42-136.84, 0.65-26.07, and 1.10-44.17 μg·mL-1 (r<0.9993), respectively. The RSD values of precision, reproducibility, and sample stability were not more than 2.2%. The average recoveries of the six components were 99.8%-101.1% with RSDs not more than 1.2%. The P values of external standard method and QAMS by paired t-test were greater than 0.05. CONCLUSION: There is no significant difference in the content analysis results of the two methods, which can both used for simultaneous determination of the four iridoids and two flavonoids in Hedyotis diffusa Willd.
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Objective To study the antitumor activities and immunity regulation of Hedyotis diffusa Willd injection. Methods Inhibitory effects of Hedyotis diffusa Willd injection on cell proliferation was detected with MTT method. Then the inhibitive rate was calculated. The immunomdulatory effects of Hedyotis diffusa Willd injection were investigated. Results Hedyotis diffusa Willd injection inhibited the proliferation of A549, SGC-7901, HEP-G2, Helacells in the dose of 100～20 ?g/mL, and enhance the immunity regulation on S180 tumor transplanted in mice. Conclusion Hedyotis diffusa Willd injection has the antitumor activities and can enhance the immunity.
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Objective To study the inducing apoptosis of Hedyotis diffusa extract (HDE) on human hepatocarcinoma cell line Bel7402 and its molecular mechanism. Methods Human hepatocarcinoma cell line Bel7402 was used in vitro cell culture, there were 3 groups:control group, HDE group and 5-Fu group, the Bel7402 cells were dealed with them respectively. The apoptosis morphologic changes of human hepatocarcinoma cells were observed by invert microscope. The apoptosis rate was detected with hematoxylin stain by light microscope. Inhibition of Bel7402 cell proliferation was measured by MTT assay. The expression of oncogene Bcl-xl and anti-oncogene p53 of Bel7402 ceils were observed with RT-PCR assay. Results Compared with the control group, the condensation of nuclear, vacuolar degeneration of mitochondria could be found through light microscope. The apoptosis rate of HDE group was remarkably increased compared with that in control group (P