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ObjectiveTo explore the mechanism of Xueniao capsule in the treatment of acute pyelonephritis (APN) by network pharmacology and experimental verification. MethodThe effect of Xueniao capsule on APN was investigated based on the APN model in rats. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Chemistryl Database, and SymMap were searched for the chemical components of Smilacis Chinae Rhizoma,Coicis Semen, and Trachycarpi Petiolus. The target information of the components was collected from PharmMapper and SwissTargetPrediction, and disease target information from Therapeutic Target Database (TTD), DrugBank, DisGeNET, GeneCards, and Online Mendelian Inheritance in Man(OMIM). The key genes of Xueniao capsule for APN underwent Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses by Metascap. Real-time quantitative polymerase chain reaction (PCR) and Western blot were employed to verify the prediction results. ResultCompared with the blank group and the sham operation group, the model group showed an increased ratio of the left kidney to the right kidney and organ index(P<0.05, P<0.01),up-regulated white blood cells (WBC),neutrophils (NEUT),monocytes (MONO), and lymphocytes (LY)(P<0.05, P<0.01), and elevated levels of nuclear factor-κB(NF-κB), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)(P<0.05, P<0.01). Compared with the model group, the norfloxacin group, the low- and high-dose Xueniao capsule groups showed a decreased ratio of the left kidney to the right kidney and organ index(P<0.05, P<0.01), dwindled levels of WBC, NEUT, MONO, and LY(P<0.05, P<0.01), and reduced levels of NF-κB, IL-6, and TNF-α(P<0.05, P<0.01). The medium-dose Xueniao capsule group showed a decreased ratio of the left kidney to the right kidney and organ index(P<0.05, P<0.01), reduced levels of WBC, NEUT, MONO, and LY(P<0.05, P<0.01), and dwindled levels of IL-6 and TNF-α(P<0.05, P<0.01). Network pharmacological analysis revealed 17 active compounds from Smilacis Chinae Rhizoma, 18 active compounds from Coicis Semen, six active compounds from Trachycarpi Petiolus, and 39 key genes for the treatment of APN in Xueniao capsule. GO enrichment analysis demonstrated 704 biological processes, 22 cellular components, and 59 molecular functions. Sixty-two pathways were enriched in KEGG enrichment analysis. The experimental verification results showed that compared with the blank group, the model group showed increased mRNA expression of prostaglandin-endoperoxide synthase 2 (PTGS2), mitogen-activated protein kinase 1 (MAPK1)/extracellular signal-regulated protein kinase 2 (ERK2),phosphoinositide 3 kinase (PI3K),protein kinase B2(Akt2),Janus kinase 2 (JAK2),and signal transducer and activator of transcription 3 (STAT3)and protein expression of PI3K, Akt2, JAK2, and STAT3 (P<0.05, P<0.01). Compared with the model group, the low-dose Xueniao capsule group showed decreased mRNA expression of MAPK1, PI3K, JAK2, and STAT3 and protein expression of PI3K, JAK2, and STAT3 (P<0.05, P<0.01). The medium-dose Xueniao capsule group showed decreased mRNA expression of MAPK1, PTGS2, PI3K, JAK2, and STAT3, and protein expression of PI3K, JAK2, and STAT3 (P<0.05, P<0.01). The high-dose Xueniao capsule group showed reduced mRNA expression of PTGS2, MAPK1, PI3K, Akt2, JAK2, and STAT3 and protein expression of PI3K, Akt2, JAK2, and STAT3 (P<0.05, P<0.01). ConclusionXueniao capsule has a certain curative effect on APN via multiple targets and multiple pathways. The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway and the JAK2/STAT3 signaling pathway.
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To evaluate the therapeutic effect of Potentilla discolor on 2,4,6-trinitrobenzensulfonic acid(TNBS)-induced experimental ulcerative colitis(UC) in rats and to determine its therapeutic mechanism through mitochondrial autophagy, immune cells, and cytokines. A rat model of UC was established by TNBS-ethanol enema. Rats were divided into six groups: control, UC model, sulfasalazine(positive drug), and high-dose, moderate-dose, and low-dose ethanol extract groups. After 14-day continuous administration of the corresponding drugs, the disease activity index(DAI) and hematoxylin and eosin(HE) were evaluated. The morphological structure of mitochondria was observed by using transmission electron microscope(TEM), mitophagy-related mRNA expression was detected by using Real-time quantitative polymerase chain reaction(qRT-PCR), immune cell differentiation in rat serum was detected by using flow cytometry(FCM), and cytokine expression in colon tissues of rats was detected by protein microarray. The results showed that compared with the model group, each dose group of P. discolor could significantly reduce the DAI of UC model rats, and decrease the degree of inflammatory cells infiltration in the colon tissue of UC model rats. Meanwhile the expressions of T cells and Th cells in the serum increased significantly, the expression of Tc cells in the serum decreased significantly. Transmission electron microscope found that there was fusion of mitochondria and lysosomes in the colon tissue of the administration group. The expressions of mitochondrial autophagy related genes NF-κB, p62 and parkin were significantly increased in colon tissues. The results of protein chip showed that compared with the model group, the high dose group of P. discolor could significantly regulate the expression of cytokines. In conclusion, these results suggested that P. discolor improved TNBS-induced acute ulcerative colitis in rats by regulating the mitochondrial autophagy and the inflammatory factor expression.
Subject(s)
Animals , Rats , Autophagy , Colitis, Ulcerative/genetics , Colon , Mitochondria , Potentilla/geneticsABSTRACT
The effects of hemolysis and lipemia on thromboelastography (TEG) analysis have been scarcely evaluated in human samples, and neglected in clinical practice. We aimed to investigate the effects of in vitro mechanical hemolysis and lipemia on TEG analysis and conventional coagulation tests. Twenty-four healthy volunteers were enrolled in the study. Besides the controls, three groups with slight, moderate and severe mechanical hemolysis were constituted according to free hemoglobin (Hb) concentrations of 0.5-1.0, 2.0-6.0 and 7.0-13.0 g/L, respectively; and three groups with mild, moderate and high lipemia were established according to triglyceride concentrations of â¼6.0, â¼12.0, and â¼18.0 mmol/L, respectively. Four TEG parameters, reaction time (R), coagulation time (K), angle (α), and maximum amplitude (MA), were measured alongside conventional plasma tests including prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FIB) by mechanical method, and platelet count by optical method. Results showed that the median R and MA values at moderate and severe hemolysis and K at severe hemolysis exceeded respective reference intervals, and were considered unacceptable. Median values of TEG parameters in lipemic samples were all within reference intervals. Bias values of conventional plasma tests PT, APTT and FIB in hemolyzed or lipemic samples were all lower than the Clinical Laboratory Improvement Amendments (CLIA) allowable limits. Bias values of platelet count at moderate to severe hemolysis and lipemia exceeded the CLIA allowable limits. In conclusion, the detection of TEG was in general more affected by mechanical hemolysis than plasma coagulation tests. Pre-analytical variables should be taken into account when unexpected TEG results are obtained.
Subject(s)
Blood Coagulation Tests/standards , Erythrocytes/chemistry , Phospholipids/chemistry , Soybean Oil/chemistry , Thrombelastography/standards , Adult , Blood Coagulation , Cells, Cultured , Emulsions/chemistry , Female , Healthy Volunteers , Hemoglobins/analysis , Hemolysis , Humans , Hyperlipidemias/blood , Kaolin , Male , Models, Biological , Platelet Count , Thrombelastography/instrumentation , Triglycerides/analysisABSTRACT
This paper was aimed to investigate the effect of Aralia echinocaulis containing serum on expression of β-catenin, Wnt-1, Frizzed-2, TCF and Axin in Wnt/β-catenin signaling pathway of primary osteoblasts. SD healthy female rats (n=80) were used to make A. echinocaulis containing serum by gastric perfusion for seven days with distilled water, A. echinocaulis decoction high dosage, middle dosage, and low dosage. In vitro, primary osteoblasts were cultured and identified. The third generation primary osteoblasts were taken and cultured for 48 h, then cells were treated with the different drug serums for 10 days and calcified nodules were counted by alizarin red staining. The cells were collected after treatment for 48 h and the expression levels of β-catenin, Wnt-1, Frizzled-2, TCF and Axin were detected by Real-time PCR and Western blot. The results suggested that the in vitro cells were primary osteoblasts; and after treatment, various doses groups could promote the mineralization ability of primary osteoblasts, up-regulate the mRNA and protein expression levels of β-catenin, Wnt-1, Frizzled-2, and TCF, and down-regulate the mRNA and protein expression levels of Axin. These findings indicated that A. echinocaulis containing serum can enhance the differentiation and proliferation of osteoblasts by regulating the expression levels of β-catenin, Wnt-1, Frizzled-2, TCF and Axin in Wnt/β-catenin signaling pathway of primary osteoblasts.
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The solubility and permeability on four kinds of flavonoids (kaempferol, hesperidin, apigenin, genistein) were test according to the theory of biopharmaceutics classification system (BCS), and their absorption mechanism. The solubility was investigated by the method in determination of solubility of "Chinese Pharmacopoeia 2010". To detect appearance permeability of compounds mentioned above, the appropriate concentrations were selected by the MTT method in cell transfer experiments in Caco-2 cell model, which established by in vitro cell culture method. Therefore, these compounds were classified with BCS according to solubility and permeability. In addition, to explore absorption mechanisms, the experiments in three different concentrations of compounds in high, medium and low in bidirectional transformation methods in Caco-2 cell model contacted. The study indicated that all of kaempferol, hesperidin, apigenin, genistein have the characteristics in low solubility and high permeability, which belong to BCSⅡ, and the absorption mechanism of kaempferol was active transportation. Whereas, hesperidin, apigenin, genistein were passive transportation. In this study, it carried out initial explorations on establishment of determination for solubility and permeability in flavonoids, and provided theoretical reference for further research on BCS in traditional Chinese medicine.
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<p><b>OBJECTIVE</b>To study and evaluate the effect of Sangu Decoction (SGD, ) on the bone destruction due to mammary cancer metastasis.</p><p><b>METHODS</b>Metastasis rat mammary tumor-1 cells were transplanted into the left hind limb tibia of SD rats to establish the bone metastasis of the mammary cancer model. The modeled rats were treated with SGD for observing its effect on rats' pain behavior, including 50% paw withdrawal threshold (50% PWT) after von Frey fiber stimulation, burden difference of bilateral feet, and thermal withdrawal latency (TWL), with zoledronic acid as the positive control. Moreover, the damage in the tibia sample of rats was scored by an iconographic method, and the bone mineral density (BMD) as well as the bone mineral content (BMC) were estimated.</p><p><b>RESULTS</b>The model established showed characteristics of mixed metastasis, revealing the manifestations of tumor development, bone destruction, cancerous pain, etc. In the SGD-treated group, 50% PWT was prolonged (8.13 ± 4.76 vs. 2.30 ± 2.19), and TWL was longer (3.48 ± 0.62 s vs. 2.89 ± 0.26 s) than those in the control group, respectively (P<0.05 or P<0.01). Iconographic scoring also showed improvement of BMD (0.134 ± 0.009 vs. 0.120 ± 0.007, P<0.01) and an elevating trend of BMC in the SGD-treated group.</p><p><b>CONCLUSION</b>SGD could effectively alleviate the cancerous pain of bone metastasis and mitigate the metastasis that cause osteolytic destruction of bone.</p>
Subject(s)
Animals , Female , Rats , Bone Density , Bone Neoplasms , Drug Therapy , Bone and Bones , Pathology , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Mammary Neoplasms, Animal , Pathology , Rats, Sprague-Dawley , Reaction TimeABSTRACT
<p><b>OBJECTIVE</b>To explore the correlation between excision repair cross-complementing 1 (ERCC1) C8092A and C19007T gene polymorphisms and different Chinese medicine (CM) syndrome types of colorectal cancer (CC).</p><p><b>METHODS</b>Ninety-nine patients with CC were syndrome typed as dampness-heat accumulation syndrome, qi stagnation with blood stasis syndrome, Pi-Shen yang deficiency syndrome, and Gan-Shen yin deficiency syndrome. The gene polymorphisms of excision repair cross-complementing 1 (ERCC1) C8092A and C19007T in different CM syndrome types of CC were examined by polymorphisms chain reaction amplification and direct sequencing, and analyzed statistically.</p><p><b>RESULTS</b>The frequencies of C8092A genotype and allele in different CM syndrome types had no statistical difference (P > 0.05). The frequencies of C19007T genotype and allele in different CM syndrome types had statistical difference (P < 0.05). Of them, there was no statistical difference in the frequencies between dampness-heat accumulation syndrome and qi stagnation with blood stasis syndrome, or between Pi-Shen yang deficiency syndrome and Gan-Shen yin deficiency syndrome (P > 0.05). There was statistical difference between dampness-heat accumulation syndrome and Pi-Shen yang deficiency syndrome as well as Gan-Shen yin deficiency syndrome (P < 0.05). There was statistical difference between qi stagnation with blood stasis syndrome and Pi-Shen yang deficiency syndrome as well as Gan-Shen yin deficiency syndrome (P < 0.05).</p><p><b>CONCLUSION</b>ERCC1 C19007T gene polymorphisms might be associated with CM syndrome types of CC, which needed to be further studied.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Colorectal Neoplasms , Diagnosis , Genetics , DNA-Binding Proteins , Genetics , Endonucleases , Genetics , Genotype , Medicine, Chinese Traditional , Polymorphism, Genetic , Yang Deficiency , Yin DeficiencyABSTRACT
<p><b>OBJECTIVE</b>To identify the tyrosinase inhibitory constituent quickly from Potentilla bifurca.</p><p><b>METHOD</b>The active constituent was found through fraction collecting and tyrosinase inhibitory activity by bioassay-linked HPLC method.</p><p><b>RESULT</b>The methanol extracts and BuOH fraction of Potentilla bifurca showed strong tyrosinase inhibitory activities. From BuOH fraction of Potentilla bifurca, the tyrosinase inhibitory constituent was isolated and identified as flavonoid, quercetin-4'-O-beta-D-glucoside. It express stronger tyrosinase inhibition than the known tyrosinase inhibitor, kojic acid (IC50 = 0.28 mmol x L(-1)) with IC50 value of 0.001 9 mmol x L(-1).</p><p><b>CONCLUSION</b>Bioassay-linked HPLC fractionation method was provided for determination the active constituents quickly from herbal medicines.</p>
Subject(s)
Enzyme Inhibitors , Chemistry , Kinetics , Peptides , Chemistry , Plant Extracts , Chemistry , Potentilla , ChemistryABSTRACT
Chicory (Cichorium intybus L) is a bushy perennial herb with blue, lavender, or occasionally white flowers. It grows as a wild plant on roadsides in its native Europe, and in North America, where it has become naturalized. Common chicory is also known as blue sailors, succory, and coffeeweed. Chicory contains saccharides, organic acid, alkaloid, triterpenes, sesquiterpenes, coumarins, and so on. It has a function of lowering the blood glucose and lipid, decreasing uric acid, and hepatoprotection. Therefore, it is evacuant and appetitive with better cardiovascular effect. Furthermore, it can be sorbefacient calcium, enhancing immunity via antiallergic, antibacterial and antivirus. So, with research and development on the peculiar physiology function of chicory, it must have a bright prospect on discovering salubrious beverage, functional food and remedy with chicory at present and near future.
Subject(s)
Animals , Humans , Cichorium intybus , Chemistry , Drug Therapy , Plant Extracts , Chemistry , Pharmacology , ResearchABSTRACT
The time of six meridians diseases tending to be cured in Shang Han Lun (Treatise on Cold Diseases) is always one of the key points for study, but up till now no satisfied explanations are made. The authors try to study it on the basis of the theory of "three-yin and three-yang" according to the relationship between human body and the environment.
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<p><b>AIM</b>To study the anti-tumor bioactive steroid saponins of Paris vietnamensis (Takht.).</p><p><b>METHODS</b>The constituents were isolated and purified by chromatography and their structures were identified by spectral analysis and physicochemical properties. The cytotoxic bioactivities of the constituents were determined by MTT.</p><p><b>RESULTS</b>Eleven compounds were obtained and identified as 3beta, 5alpha,6alpha-trihydroxy-7(8)-en-isospirostanol-3-O-beta-D-glucopyranosyl(1-->3) [alpha-L-rhamnopyranosyl(1-->2)]-beta-D-glucopyranoside (1), which was named as parisvietnaside A, 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (2), 25(R) diosgenin-3-O-alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (3), 25 (R) diosgenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (4), 25 (R) diosgenin-3-O-beta-D-glucopyranosyl (1-->3) [alpha-L-rhamnopyranosyl (1-->2)]-beta-D-glucopyranoside (5), 25 (R) diosgenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (6), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl(1-->4)-beta-D-glucopyranoside (7), 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranoside (8), 25 (R) pennogenin-3-O-alpha-L-arabinofuranosyl (1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (9), 25 (R) pennogenin-3-O-beta-D-glycopyranosyl (1-->3) [alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranoside (10) and 25 (R) pennogenin-3-O-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl(1-->4) [alpha-L-rhamnopyranosyl-(1-->2)]-beta-D-glucopyranoside (11). Some constituents had cytotoxic bioactivities.</p><p><b>CONCLUSION</b>Compound 1 is a new spirostanol saponin, and compounds 2, 3, 6-11 were obtained from Paris vietnamensis (Takht.) for the first time. Compounds 3, 4, 6, 8 had cytotoxic bioactivities against tumor cells HepG2 and SGC-7901.</p>
Subject(s)
Humans , Adenocarcinoma , Pathology , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Cell Proliferation , Liliaceae , Chemistry , Liver Neoplasms , Pathology , Molecular Conformation , Molecular Structure , Plants, Medicinal , Chemistry , Rhizome , Chemistry , Saponins , Chemistry , Pharmacology , Stomach Neoplasms , PathologyABSTRACT
Objective:To evaluate the anti-metastatic and bone preserving therapeutic effects of herbal medicine extraction on rat model of bone metastatic carcinoma.Methods:A rat model of cancer-induced bone pain using the MRMT-1 cell line injected into the tibia was established to investigate the efficacy of the herbal medicine extraction,on osteoclast activity and bone mineral density.The development of the bone tumor and structural damage to the bone was monitored by radiological analysis.Specimens of the tibial bone were processed for tartrate-resistant acid phosphatase(TRAP)stain to observe the bone pathological changes and count TRAP stained osteoclasts.OPG and RANKL expression was evaluated by immunohistological methord.Results:Histological and radiological examination showed that the herbal medicine extraction significantly inhibited tumor proliferation and preserved the cortical and trabecular bone structure.In addition,a dramatic reduction of tartrate resistant acid phosphatase-positive polykaryocytes(osteoclasts)and increase of OPG expression were observed.Conclusions:The herbal medicine extraction was an anti-metastatic and bone preserving therapeutic effects in a rat model of metastatic cancer pain.
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The collected information is an attempt to cover the more recent developments in the phytochemistry and pharmacology of this genus. During the past years, alkaloids, flavonoids, volatile oils, organic acids, polysaccharides, tannins and phenolic constituents have been isolated from Ephedra. Pharmacological studies are described according to hypoglycemic effects, anticoagulated blood properties, depressurization, immunosuppressive activity, antioxidation and antivirus activity and so on. The information summarized here is intended to provid a rational foundation for the futher development and utilization of Ephedra which is rich in China.
Subject(s)
Animals , Alkaloids , Chemistry , Ephedra , Chemistry , Classification , Flavonoids , Chemistry , Hypoglycemic Agents , Pharmacology , Immunosuppressive Agents , Pharmacology , Molecular Structure , Plant Stems , Chemistry , Plants, Medicinal , Chemistry , Polysaccharides , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>The several species of the genus Paris called as Rhizoma Paridis were famous traditional Chinese medica. To develop the quantitative analysis method of the steroidal saponins in some species of the genus Paris and commercially available Rhizoma Paridis samples by HPLC-ELSD.</p><p><b>METHOD</b>The contents of 11 steroidal saponins in Rhizoma Paridis samples were dectected with a Kromasil C18(4.6 mm x 150 mm, 5 microm) column which was deluted with acetonitrile-water (30:70-60:40) at a flow rate of 1 mL x min(-1) by HPLC-ELSD.</p><p><b>RESULT</b>All the authentic samples could be separated and calibration curves of 11 saponins were prepared. 11 steroidal saponins in 16 Rhizoma Paridis samples were detected in 30 min. The recovery for the assay of saponins was between 95% and 97%. The precision and stability of samples (RSD) were below 3%.</p><p><b>CONCLUSION</b>The method was shown to be accurate and convenient, and suitable for the quantitative analysis of these 11 steroidal saponins in the commercially available Rhizoma Paridis samples.</p>