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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 187-192, 2019.
Article in Chinese | WPRIM | ID: wpr-802543

ABSTRACT

Objective: To identify the chemical compositions and conduct relative content analysis of the essential oil from wild Doellingeria scaber in high altitude area of Dabie mountains, and investigate its antioxidant capacity. Method: The volatile oil of D. scaber was extracted by simultaneous distillation extraction (SDE) method. Its chemical compositions were analyzed by gas chromatography-mass spectometry (GC-MS), and the relative percentage content of each composition was determined by peak area normalization method. The antioxidant activities of the oil were evaluated by reducing ability method, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) method and β-carotene bleaching assay. Result: A total of 50 components representing 86.91% in this plant were identified. The main chemical compositions were (E)-β-farnesene(20.21%), germacrene D(9.94%), hexadecanoic acid(8.66%), β-terpinene(7.82%), caryophyllene(6.9%) and p-cymen-8-ol(4.48%). The main components of volatile oils include alkenes (56.57%), alcohols (12.24%), fatty acids (11.24%), epoxides(2.93%), esters(1.82%), aldehydes and ketones (1.60%), and aromatics (0.51%). The reduction ability of volatile oil to iron ions increases with the increase of concentration, but the reduction ability is far weaker than that of vitamin C. The 50% inhibiting concentration (IC50) of volatile oil on DPPH free radicals and in β-carotene/linoleic acid lipid system was 0.72 g·L-1 and 0.10 g·L-1, respectively, indicating that the volatile oil had a certain scavenging capacity on DPPH free radicals and a good inhibition effect on lipid peroxidation. Conclusion: GC-MS was used to identify the chemical compositions of the volatile oil from D. scaber, and antioxidant test in vitro showed that D. scaber had certain antioxidant capacity, providing experimental basis for its development and utilization.

2.
Chinese journal of integrative medicine ; (12): 137-142, 2013.
Article in English | WPRIM | ID: wpr-347114

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of Qushuanling Capsule ( QSLC) on thrombus formation and platelet aggregation in rats.</p><p><b>METHODS</b>Arteriovenous bypass, venous thrombosis, and middle cerebral artery thrombosis models were used in rats to investigate the anti-thrombotic effects of QSLC, a compound of nine Chinese herbs. The platelet aggregation induced by adenosine diphosphate (ADP), thrombin or arachidonic acid (AA), as well as the contents of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1α (6-keto-PGF1α) in rat plasma and aortic walls, were determined to investigate the possible mechanisms of the anti-thrombotic effects of QSLC.</p><p><b>RESULTS</b>After oral administration with QSLC for 7 days, arteriovenous bypass thrombosis was obviously suppressed compared with the model group, venous thrombosis was also obviously suppressed, rat behaviors were obviously improved, and brain infarct size as well as water content were also reduced. The platelet aggregation induced by ADP or thrombin was inhibited by QSLC, but the drug had no effect on AA-induced platelet aggregation and content of TXB(2) and 6-keto-PGF1α in plasma and the aortic wall.</p><p><b>CONCLUSION</b>These results suggest that QSLC can be used in the prevention and treatment of thrombotic diseases, and that its mechanism of action may be related to inhibition of platelet aggregation.</p>


Subject(s)
Animals , Male , Rats , 6-Ketoprostaglandin F1 alpha , Blood , Adenosine Diphosphate , Pharmacology , Aorta , Metabolism , Pathology , Cerebral Infarction , Blood , Drug Therapy , Pathology , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Middle Cerebral Artery , Pathology , Platelet Aggregation , Rats, Sprague-Dawley , Thrombosis , Drug Therapy , Pathology , Thromboxane B2 , Blood , Venous Thrombosis , Drug Therapy , Pathology
3.
Asian Pacific Journal of Tropical Medicine ; (12): 891-896, 2012.
Article in English | WPRIM | ID: wpr-819573

ABSTRACT

OBJECTIVE@#To investigate the effects of puerarin on the activity of superoxide dismutase (SOD), and expressions of advanced glycation end-product (AGE) receptor (RAGE) and vascular endothelial growth factor (VEGF) in retinas of streptozotocin (STZ)-induced early diabetic rats.@*METHODS@#Diabetic rat models were established by inducing diabetes via intra-peritoneal injection of STZ. Rats were randomly divided into normal (control), diabetic (DM), and DM+ puerarin groups. After intra-gastric administration of puerarin (500 mg/kg/day for 4 weeks), levels of SOD and malondialdehyde (MDA) were determined in serum and retina. mRNA and protein expression levels of RAGE and VEGF in retinas were determined by real-time polymerase chain reaction (RT-PCR) (mRNA) and Western blot analysis (protein levels).@*RESULTS@#There was significantly lower SOD activity and significantly higher MDA in serum and retinas of the DM group compared with the two other groups (P<0.05). After treatment with puerarin, SOD activity increased and MDA content decreased in this group (P<0.05). mRNA and protein expression levels of RAGE and VEGF in the DM group were significantly higher than those of the other groups (P<0.05), and decreased after puerarin treatment (P<0.05).@*CONCLUSIONS@#Puerarin is able to enhance SOD activity, and inhibit RAGE and VEGF expressions in retinas of STZ-induced early diabetic rats.


Subject(s)
Animals , Male , Rats , Blotting, Western , Diabetes Mellitus, Experimental , Drug Therapy , Pathology , Enzyme Activators , Gene Expression Profiling , Isoflavones , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptor for Advanced Glycation End Products , Receptors, Immunologic , Retina , Pathology , Superoxide Dismutase , Metabolism , Treatment Outcome , Vascular Endothelial Growth Factor A
4.
Acta Pharmaceutica Sinica ; (12): 1006-1011, 2010.
Article in Chinese | WPRIM | ID: wpr-353393

ABSTRACT

This study is to investigate the therapeutic effect of syringin on adjuvant arthritis (AA) in rats and its mechanisms. Complete Freund's adjuvant (FCA) was used to induce AA in rats. Secondary paw swelling of AA rats was measured with volume meter. Pain response and polyarthritis index were scored. Meanwhile, splenic lymphocyte proliferation response induced by concanavalin A (ConA) or lipopolysaccharide (LPS) was examined with MTT assay. IL-2 production of splenic lymphocytes and IL-1 beta, TNF-alpha production of peritoneal macrophage (PM phi) were estimated by enzyme linked immunosorbent assay (ELISA). The secondary inflammation of AA rats appeared on the 14th day after injection of FCA. Syringin and tripterygium glycosides (TG) were given by intragastric administration for 16 days from the 14th day. Treatment of AA rats with syringin and TG from the 22th day significantly attenuated the secondary hind paw swelling, as well as relieved the pain response and the polyarthritic symptoms of the whole body as compared with that of the AA model group. The suppressed lymphocyte proliferation and IL-2 production of splenic lymphocytes in AA rats were reversed by treatment with syringin. Meanwhile, syringin remarkably down-regulated IL-1 beta, TNF-alpha productions from PM phi. These results indicate that anti-inflammatory effects of syringin on AA rats are mediated by modulating the immune function of abnormal cells and the balance of cytokines.


Subject(s)
Animals , Male , Rats , Anti-Inflammatory Agents , Pharmacology , Arthritis, Experimental , Drug Therapy , Metabolism , Pathology , Cell Proliferation , Eleutherococcus , Chemistry , Freund's Adjuvant , Glucosides , Pharmacology , Interleukin-1beta , Metabolism , Interleukin-2 , Metabolism , Lymphocytes , Metabolism , Pathology , Macrophages, Peritoneal , Metabolism , Phenylpropionates , Pharmacology , Plants, Medicinal , Chemistry , Random Allocation , Rats, Wistar , Spleen , Pathology , Tumor Necrosis Factor-alpha , Metabolism
5.
Acta Pharmaceutica Sinica ; (12): 737-740, 2009.
Article in Chinese | WPRIM | ID: wpr-278190

ABSTRACT

This study is to investigate the effect of ibudilast on apoptosis of airway eosinophil in asthmatic guinea pigs and its mechanism. Experimental asthma model of guinea pigs was induced with ovalbumin (OVA). Differential count in BALF was examined. The apoptosis of eosinophils (EOS) was labeled with TdT-mediated dUTP nick end labeling (TUNEL) technique. Fas mRNA expression of EOS was detected by reverse transcription-polymerase chain reaction (RT-PCR). The quantification of GM-CSF and IL-5 in BALF was conducted with ELISA. After treatment of ibudilast, the number of EOS and the quantification of GM-CSF and IL-5 decreased significantly. The number of apoptotic cells as well as Fas mRNA expression of EOS obviously increased. The results indicated that anti-asthma mechanisms of ibudilast can antagonize asthma through decreasing the number of EOS, inducing apoptosis of EOS, enhancing Fas mRNA expression of EOS and reducing the content of GM-CSF and IL-5.


Subject(s)
Animals , Female , Male , Anti-Asthmatic Agents , Pharmacology , Apoptosis , Asthma , Metabolism , Pathology , Bronchoalveolar Lavage Fluid , Eosinophils , Cell Biology , Metabolism , Granulocyte-Macrophage Colony-Stimulating Factor , Metabolism , Guinea Pigs , Interleukin-5 , Metabolism , Pyridines , Pharmacology , fas Receptor , Metabolism
6.
Journal of Southern Medical University ; (12): 1341-1343, 2009.
Article in Chinese | WPRIM | ID: wpr-268765

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of p21-activated kinase-1 (PAK1) gene transfection on the invasiveness of human colorectal carcinoma SW480 cells in vitro.</p><p><b>METHODS</b>SW480 cells in routine culture were transfected with the recombinant plasmid EGFP-C1/PAK1 via Lipofectamine(TM) 2000. The expression of PAK1 protein in SW480 cells was detected using Western blotting, and the changes of the invasiveness of SW480 cells were evaluated using Boyden chamber invasion assay.</p><p><b>RESULTS</b>Forty-eight hours after transfection with pEGFP-C1/ PAK1, the PAK1 protein expression increased significantly in comparison with those in negative and vector control groups. The invasiveness of the SW480 cells was significantly enhanced after the transfection.</p><p><b>CONCLUSION</b>The PAK1 gene transfection can increase the expression of PAK1 in SW480 cells and enhance the invasiveness of the cells. PAK1 can be associated with the invasiveness and metastasis of colorectal carcinoma cells.</p>


Subject(s)
Humans , Cell Line, Tumor , Colorectal Neoplasms , Genetics , Pathology , Gene Expression , Genetic Vectors , Neoplasm Invasiveness , Neoplasm Metastasis , Plasmids , Transfection , p21-Activated Kinases , Genetics
7.
China Journal of Chinese Materia Medica ; (24): 769-821, 2008.
Article in Chinese | WPRIM | ID: wpr-284398

ABSTRACT

<p><b>OBJECTIVE</b>To design a macroporous resin chromatography system with on-line detector which can be used to purify total flavonoid glycosides from Hypericum perforatum.</p><p><b>METHOD</b>Compared with the present detection methods, the ultraviolet detector which can be applied to flow liquid, and a self-made shunt were applied to design the on-line detection system. This system was successfully applied to purify the total flavonoid glycosides from H. perforatum.</p><p><b>RESULT</b>This on-line detection system was simple and feasible, and the results were accurate and reliable. The purity of total flavonoid glycosides from H. perforatum. separated by this system was higher than 90%.</p><p><b>CONCLUSION</b>This technique provides a simple and reliable on-line detection method for industrial chromatography purification of TCM extracts, which is the basis of automatic manufactory of TCM purification.</p>


Subject(s)
Adsorption , Chromatography , Flavonoids , Chemistry , Glycosides , Chemistry , Hypericum , Chemistry , Online Systems , Porosity , Quality Control , Reproducibility of Results , Resins, Plant , Chemistry , Sensitivity and Specificity , Ultraviolet Rays
8.
China Journal of Chinese Materia Medica ; (24): 1865-1868, 2008.
Article in Chinese | WPRIM | ID: wpr-252206

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of nebulized total ginkgo flavone glycosides (TFG) on Th1/Th2 imbalance in mice with athma.</p><p><b>METHOD</b>Forty-eight BALB/C mice were randomly divided into four groups: group A (control group, n=12); group B (asthmatic model group, n=12); group C (TFG nebulized treated group, n=12); group D (dexamethasone intraperitoneal treated group, n=12). The asthmatic model was established by sensitivity and local activation with Ovalbumin(OVA) and aluminum hydroxide Al(OH)3. TFG (50 g x L(-1), per aerosol per six mice, 30 minutes) was nebulized 20 days after modeling, while dexamethasone (1 g x L(-1)) was intraperitoneal once daily for 10 days. Perfusate of bronchoalveolar lavage fluid(BALF) was collected on day 32. The level of IL-4, IFN-gamma in BALF, and the level of total IgE in serum was determined. The airway inflammation pathology change and the expression of GATA-3 protein in lungs was detected by immunohistochemical staining.</p><p><b>RESULT</b>Compared with model group, the decreased content of IL-4(49.30 +/- 7.95) ng x L(-1) and increased level of IFN-gamma (49.08 x 5.46) ng x L(-1). were found in BALF, and the level of total IgE (9.47 +/- 1.52) microg x L(-1) in serum also decreased in TFG treated group. In model group, smooth muscle hypertrophing, mucous hyperemia, mucous layer thickening, and inflammatory cell in filtration were observed. Phlegmasia was appeared in the bronchi, which was filled with lots of mucus. In contrast, the inflammatory reaction in TFG and Dexamethasone treated group was less obvious. Expression of GATA-3 was markly increased in model group with decreased expression in TFG treated group.</p><p><b>CONCLUSION</b>Nebulized TFG showed a therapeutic effect for asthmatic mice, the mechanism may be explained by blockingnnnn GATA-3 expression and regulating the disorder Th1/Th2 imbalance.</p>


Subject(s)
Animals , Female , Male , Mice , Aluminum Hydroxide , Pharmacology , Asthma , Drug Therapy , Metabolism , Pathology , Bronchoalveolar Lavage Fluid , Chemistry , Dexamethasone , Therapeutic Uses , Drugs, Chinese Herbal , Chemistry , Therapeutic Uses , Flavones , Flavonoids , Chemistry , GATA3 Transcription Factor , Metabolism , Ginkgo biloba , Chemistry , Glycosides , Chemistry , Therapeutic Uses , Immunoglobulin E , Blood , Interferon-gamma , Metabolism , Interleukin-4 , Metabolism , Lung , Metabolism , Mice, Inbred BALB C , Ovalbumin , Pharmacology , Random Allocation , Th1 Cells , Metabolism , Th2 Cells , Metabolism
9.
Journal of Southern Medical University ; (12): 915-918, 2008.
Article in Chinese | WPRIM | ID: wpr-280069

ABSTRACT

<p><b>OBJECTIVE</b>To clone migfilin-N terminal sequence into E.coli and obtain a fusion protein for preparing rabbit polyclonal antibody against migfilin, thereby facilitating the study of the role of migfilin in the biological behavior of colon cancer.</p><p><b>METHODS</b>Based on human migfilin cDNA sequence, a pair of primers was designed to amplify migfilin-N terminal sequence by PCR. The PCR product was subcloned into the bacterial expression vector pGEX-4T-1 with EcoRI/XhoI sites, and the target recombinant plasmids were identified with enzymatic cleavage followed by DNA sequence analysis. By transforming the expression vector into component E.coli BL(21) cells, the GST-migfilin-N fusion protein was expressed with IPTG induction. Glutathione-sepharose beads were used to purify the fusion protein, and anti-migfilin polyclonal antibody was produced by immunization of rabbits with the purified GST-migfilin N-terminal fusion protein. The resultant anti-migfilin polyclonal antibody was purified by protein A beads and used for Western blotting for detecting migfilin expression in different cell lines.</p><p><b>RESULTS AND CONCLUSION</b>The migfilin-N terminal gene fragment was cloned successfully, and purified GST-migfilin N-terminal fusion protein and anti-rabbit migfilin polyclonal antibodies were obtained. Western blot analysis demonstrates that the antibodies specifically detected migfilin expression in the cell lines, which may facilitate further investigation of the role of migfilin in the biology of colon cancer.</p>


Subject(s)
Animals , Humans , Rabbits , Antibodies, Monoclonal , Allergy and Immunology , Base Sequence , Blotting, Western , Cell Adhesion Molecules , Genetics , Allergy and Immunology , Cell Line, Tumor , Cloning, Molecular , Colonic Neoplasms , Genetics , Metabolism , Pathology , Cytoskeletal Proteins , Genetics , Allergy and Immunology , DNA, Complementary , Chemistry , Genetics , Escherichia coli , Genetics , Molecular Sequence Data , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
10.
Acta Pharmaceutica Sinica ; (12): 480-483, 2008.
Article in Chinese | WPRIM | ID: wpr-277827

ABSTRACT

This study was to investigate the effect of total flavonoid in leaves of Ginkgo biloba (total flavonoid in leaves of Ginkgo biloba, FG) on the apoptosis of eosinophils (EOS) in broncho alveloar lavage fluid (BALF) of asthma mice. Mouse asthma model was established by ovalbumin (OVA) challenge methods. After atomizing therapy for two weeks, differential count in BALF, morphological change and proportion of apoptosis were detected by AO/EB stain and Annexin V-FITC/PI. The number of total leucocytes and eosinophils in BALF decreased obviously after FG treatment. Compared with model group, the number and proportion of EOS apoptosis increased significantly after FG treatment. The results indicated that one of the anti-inflammation mechanisms of FG might be promoting apoptosis of eosinophils.


Subject(s)
Animals , Female , Mice , Anti-Inflammatory Agents, Non-Steroidal , Pharmacology , Apoptosis , Asthma , Pathology , Bronchoalveolar Lavage Fluid , Cell Biology , Eosinophils , Pathology , Flavonoids , Pharmacology , Ginkgo biloba , Chemistry , Leukocyte Count , Mice, Inbred BALB C , Ovalbumin , Plant Leaves , Chemistry , Random Allocation
11.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 244-246, 2007.
Article in Chinese | WPRIM | ID: wpr-269090

ABSTRACT

<p><b>OBJECTIVE</b>To study the viral inhibitory effect of Shenling Yigan Granule (SYG) on duck hepatitis B virus (DHBV) in vivo.</p><p><b>METHODS</b>Chongqing ducks infected with DHBV were used. They were randomly divided into five groups, the small-, medium- and high-dose (1.6 g/kg, 3.2 g/kg, 6.4 g g/kg) SYG groups, the lamivudine positive control group, and the model group. The changes of serum DHBV-DNA, DHB-sAg contents and hepatic pathology were observed.</p><p><b>RESULTS</b>The serum content of DHBV-DNA in the three SYG groups and the positive control group was significantly decreased (P < 0.05), while it was rebounded in the latter at day 7 after stopped lamivudine administration. The change of DHBsAg level was insignificantly in all groups. And the hepatic pathological change in the SYG groups and positive control group was slighter than that in the model control group, but showed insignificant difference in comparison between the SYG groups and the model group (P > 0.05).</p><p><b>CONCLUSION</b>SYG has certain in vivo inhibitory effects on DHBV-DNA.</p>


Subject(s)
Animals , Antiviral Agents , Pharmacology , Therapeutic Uses , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Ducks , Hepadnaviridae Infections , Drug Therapy , Hepatitis B Virus, Duck , Hepatitis, Viral, Animal , Drug Therapy , Phytotherapy , Random Allocation
12.
Journal of Southern Medical University ; (12): 827-830, 2007.
Article in Chinese | WPRIM | ID: wpr-337374

ABSTRACT

<p><b>OBJECTIVE</b>To study the role of PAK6 in prostate cancer by cloning PAK6-N terminal sequence into E.coli and preparing its polyclonal rabbit antibody to detect PAK6 expression in prostate cancer.</p><p><b>METHODS</b>Based on human PAK6 cDNA sequence, we designed a pair of primers to amplify the PAK6-N terminal sequence by PCR. The PCR product was subcloned into the bacterial expression vector pGEX-4T-1 via EcoRI/XhoI sites, and the recombinant plasmids were identified by enzymatic cleavage followed by DNA sequence analysis. By transforming the expression vector into component E.coli BL21 cells, the GST-PAK6-N fusion protein was expressed with IPTG induction. Glutathione-Sepharose beads were used to purify GST- PAK6-N fusion protein. Anti-PAK6 polyclonal antibody was produced by immunizing rabbits with purified GST-PAK6 N-terminal fusion protein. Anti-PAK6 polyclonal antibody was purified by protein A beads and used for detection of PAK6 expression in 3 prostate cancer specimens.</p><p><b>RESULTS AND CONCLUSION</b>We cloned PAK6-N terminal gene fragment successfully, purified GST-PAK6 N-terminal fusion protein, and obtained polyclonal rabbit PAK6 antibody. Immunohistochemistry indicated that PAK6 expressed in the stroma instead of the cancer cells in prostate cancer. All of the 3 prostate cancer specimens showed positive staining in the stroma, suggesting that PAK6 may participate in the stroma-cancer cell interaction in prostate cancer.</p>


Subject(s)
Aged , Animals , Humans , Male , Rabbits , Antibodies, Monoclonal , Allergy and Immunology , Cloning, Molecular , DNA, Complementary , Chemistry , Genetics , Electrophoresis, Polyacrylamide Gel , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Polymerase Chain Reaction , Prostatic Neoplasms , Genetics , Recombinant Fusion Proteins , Allergy and Immunology , Metabolism , Sequence Analysis, DNA , p21-Activated Kinases , Genetics , Allergy and Immunology , Metabolism
13.
Journal of Southern Medical University ; (12): 951-954, 2007.
Article in Chinese | WPRIM | ID: wpr-337352

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of short hairpin RNA (shRNA) targeting survivin on adhesion and invasion of human colon carcinoma cell line SW480 in vitro.</p><p><b>METHODS</b>According to the sequence of the coding region of survivin gene, two strings of 19 nucleotides of inverted sequence flanking the loop sequence of two complementary 9-base oligonucleotides were designed and synthesized to prepare the hairpin construct as the DNA templates for the target shRNA. The shRNA templates were cloned into shRNA expression vector pRNAT-U6.1/Neo, and the resulted vector pRNAT-U6.1/Neo-survivin was transfected into SW480 cells using Lipofectamine 2000. Western blotting was performed to evaluate survivin gene silencing induced by shRNA transfection at the protein level, and the biological behaviors of the SW480 cells were investigated by cell-matrix adhesion, invasion and gelatin-zymography assays.</p><p><b>RESULTS</b>Western blotting revealed significantly lowered survivin protein expression in transfected SW480 cells, and survivin gene silencing induced by shRNA significantly suppressed the metastatic potential of SW480 cells in association with suppressed MMPs activity.</p><p><b>CONCLUSIONS</b>Survivin may play an important role in modulating human colorectal carcinoma cell invasion and metastasis, and survivin gene silencing can inhibit human colorectal cancer cell invasion and the production of MMP-2 and MMP-9. Survivin may affect invasion and metastasis of human colorectal carcinoma cells via regulating the production of MMPs.</p>


Subject(s)
Animals , Humans , Blotting, Western , Cell Line, Tumor , Colonic Neoplasms , Genetics , Pathology , Gene Silencing , Inhibitor of Apoptosis Proteins , Inverted Repeat Sequences , Matrix Metalloproteinases , Bodily Secretions , Microtubule-Associated Proteins , Genetics , Neoplasm Invasiveness , Genetics , Neoplasm Metastasis , Genetics , RNA, Small Interfering , Genetics
14.
China Journal of Chinese Materia Medica ; (24): 2511-2514, 2007.
Article in Chinese | WPRIM | ID: wpr-324338

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of C21 steroidal glycoside (CSG) from the root of Cynanchum auriculatum from Jiangsu on D-galactose (D-gal) induced aging model mice.</p><p><b>METHOD</b>D-gal aging mouse model was established by cervicodorsal region subcutaneous injection with D-gal once a day for eight successive weeks. The mice in the normal control group (NCG, non-modeled) and the model control group (MCG, modeled but untreated) were treated with 1% CMC-Na. The model mice in the low, middle and high-dose CSG and Vitamin E treated groups were treated with a dose of (10, 20, 40, 100 mg x kg(-1) per day, respectively. The SOD activity, MDA content and telomerase activity in serum, heart, liver and brain tissues of mice were measured.</p><p><b>RESULT</b>CSG could obviously increase the SOD activity and decrease the MDA level in serum, heart, liver and brain tissues in D-gal aging mice (P < 0.01). There was no significant difference between three CSG treated groups and Vitamin E treated groups. In comparison of telomerase activity between MCG and the treated groups, it was shown that there was a significant increase in serum in middle and high dose group, and in heart tissues in CSG and Vit E treated groups, but was not in liver and brain tissue.</p><p><b>CONCLUSION</b>This study demonstrates that CSG can antagonize free radical injury, increase the SOD activity and decrease the MDA content of serum, heart, liver and brain in D-gal aging mice, and increase the telomerase activity in serum and heart tissues but not in liver and brain tissue.</p>


Subject(s)
Animals , Female , Male , Mice , Aging , Metabolism , Brain , Metabolism , Cynanchum , Chemistry , Drugs, Chinese Herbal , Pharmacology , Galactose , Toxicity , Glycosides , Pharmacology , Liver , Metabolism , Malondialdehyde , Blood , Metabolism , Mice, Inbred ICR , Myocardium , Metabolism , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Random Allocation , Steroids , Pharmacology , Superoxide Dismutase , Blood , Metabolism , Telomerase , Metabolism
15.
Acta Pharmaceutica Sinica ; (12): 366-370, 2007.
Article in Chinese | WPRIM | ID: wpr-281892

ABSTRACT

This study is to investigate the effect of the C21 sterols on inducing apoptosis of hepatocellular cancer cells and its potential mechanism. The transplanted model of hepatoma substantiality (Heps) was established in mice, and the mice were divided into four groups: negative controls group and C21 sterols groups (10, 20, 40 mg x kg(-1)) , treated with drugs separately once a day for 9 days. Then the mice were sacrificed, the tumor growth inhibition rate (IR) was calculated and tumor tissue samples were taken and examined under electron microscope. The tumor cells were harvested and cell viability or apoptosis was analyzed by acridine orange and ethidium bromide (AO/EB) stain. B-cell lymphoma/leukemia-2 gene (bcl-2) in tumor cells was inspected by immunohistochemistry. After treatment with C21 sterols (10, 20, 40 mg x kg(-1)), inhibitory effect on the transplanted Heps was observed. The IR was 34.79%, 47.08% and 50.23%, respectively. Apoptosis induced by the C21 sterols was observed, low growth density and some apoptotic cells were observed in tumor under the electron microscope. The expression of bcl-2 gene on tumor cells decreased in the C21 sterols groups, but the percentage of positive area is higher in 40 mg x kg(-1) group than that in 20 mg x kg(-1) group, which differed from apoptosis results. Inhibiting the excessive expression of bcl-2 gene to promote apoptosis may be one of anti-tumor mechanisms for the C21 sterols in Baishouwu.


Subject(s)
Animals , Female , Male , Mice , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Cynanchum , Chemistry , Gene Expression Regulation, Neoplastic , Genes, bcl-2 , Liver Neoplasms, Experimental , Metabolism , Pathology , Mice, Inbred ICR , Neoplasm Transplantation , Plants, Medicinal , Chemistry , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Random Allocation , Sterols , Pharmacology , Tumor Burden
16.
Journal of Southern Medical University ; (12): 349-351, 2007.
Article in Chinese | WPRIM | ID: wpr-298169

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant plasmid containing the coding region of tight junction protein claudin-1 gene to understand the functional role of claudin-1 in human colorectal carcinoma.</p><p><b>METHODS</b>The total RNA was extracted using Trizol from human colorectal carcinoma cell line SW620, and the DNA for claudin-1 was obtained by means of RT-PCR. The PCR product was inserted into the plasmid pEGFP-C1 after restriction endonuclease digestion and ligation. The recombinant plasmid was then transfected into human colorectal carcinoma cell line SW480.</p><p><b>RESULTS</b>The sequence of the recombinant plasmid was verified by restriction endonuclease and DNA sequence analysis, and the target protein expression was detected mostly on the cell membrane.</p><p><b>CONCLUSION</b>The expression vector claudin-1/pEGFP-C1 has been constructed successfully and the target protein can be expressed in human colorectal carcinoma cell line.</p>


Subject(s)
Humans , Cell Line, Tumor , Claudin-1 , Cloning, Molecular , Colorectal Neoplasms , Genetics , Metabolism , Pathology , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Membrane Proteins , Genetics , Metabolism , Microscopy, Confocal , RNA, Messenger , Genetics , Recombinant Fusion Proteins , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tight Junctions , Metabolism , Transfection
17.
Journal of Southern Medical University ; (12): 730-733, 2006.
Article in Chinese | WPRIM | ID: wpr-282931

ABSTRACT

<p><b>OBJECTIVE</b>To clone PAK5-N terminal sequence for expression in E. coli to prepare its polyclonal antibody, and examine the role of PAK5 in dental germ cells.</p><p><b>METHODS</b>Based on human PAK5 cDNA sequence, PCR primers were designed to amplify PAK5-N terminal sequence. The PCR product was cloned into the expression vector pGEX-4T-1 EcoRI/XhoI sites, and the recombinant plasmids were identified by agarose gel electrophoresis followed by DNA sequence analysis. The recombinant plasmids were transformed into E. coli BL21 and the expression of GST-fusion protein was induced by IPTG. Glutathione-Sepharose beads were used to purify GST-fusion PAK5-N-terminal fragment. Anti-PAK5 polyclonal antibody was obtained in immunizing rabbits with purified GST-PAK5 N-terminal fusion protein, and the antibodies were purified by protein A beads and used for detection of PAK5 expression in dental germ cells.</p><p><b>RESULTS AND CONCLUSIONS</b>We successfully cloned PAK5-N terminal gene fragment, and achieved protein expression, purification and production of PAK5-NT polyclonal antibody. The results of Western blotting indicated that PAK5 can be highly expressed in the dental germ cells, suggesting that PAK5 may play an important role in biological function of dental germ cells.</p>


Subject(s)
Animals , Humans , Rabbits , Antibodies, Monoclonal , Genetics , Allergy and Immunology , Base Sequence , Blotting, Western , Cloning, Molecular , Molecular Sequence Data , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Sequence Analysis, DNA , Tooth Germ , Cell Biology , Embryology , p21-Activated Kinases , Genetics , Allergy and Immunology
18.
Acta Pharmaceutica Sinica ; (12): 797-800, 2006.
Article in Chinese | WPRIM | ID: wpr-294936

ABSTRACT

<p><b>AIM</b>To study the inhibitory effect and mechanism of nobiletin on Heps tumor bearing mice.</p><p><b>METHODS</b>Models of Heps tumor bearing mice were established. The inhibitory rates of tumor growth were calculated, the apoptosis morphology of tumor tissue was observed. The T lymphocyte transformation capacity was tested by MTT assay, the TNFalpha and IL-2 production were measured by LDH kits.</p><p><b>RESULTS</b>Nobiletin could significantly inhibit Heps tumor growth. The inhibitory rates were 42.14% - 65.09% (P < 0.01). The morphology of tumor tissues in nobiletin group had typical characters of necrosis and apoptosis through transmission electron microscope. Nobiletin could stimulate T lymphocyte transformation and the production of TNFalpha and IL-2.</p><p><b>CONCLUSION</b>Nobiletin has obvious antitumor effect on Heps, the main mechanism is to enhance the cellular immune function and induce apoptosis of tumor tissue.</p>


Subject(s)
Animals , Female , Humans , Male , Mice , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Cell Line, Tumor , Cell Transformation, Neoplastic , Metabolism , Pathology , Citrus , Chemistry , Flavones , Pharmacology , Interleukin-2 , Liver Neoplasms, Experimental , Pathology , Mice, Inbred ICR , Plants, Medicinal , Chemistry , T-Lymphocytes , Metabolism , Pathology , Tumor Necrosis Factor-alpha
19.
China Journal of Chinese Materia Medica ; (24): 1425-1470, 2005.
Article in Chinese | WPRIM | ID: wpr-239656

ABSTRACT

<p><b>OBJECTIVE</b>To develop a method for the pretreatment of the series of HPD macroporous adsorption resin (MAR).</p><p><b>METHOD</b>The effect of 2% NaOH to pretreatment of the series of HPD MAR was studied by gas chromatography and UV spectrometery.</p><p><b>RESULT</b>The eluting effect by ethanol was better after the MAR was marinated and eluted by 2% NaOH.</p><p><b>CONCLUSION</b>The pretreatment of the series of HPD MAR was: after marinated and eluted by 2% NaOH, the series of HPD MAR were eluted by ethanol at 2 BV x h(-1) at 60 degrees C for 3-4 bed volumes (BV). At then the UV absorbance of eluate was 0.2-0.5. Benzene was not detected and the other residues less than 10 mg x L(-1) in eluted-MAR by GC. And the MARs match to the standard of medicine.</p>


Subject(s)
Adsorption , Chromatography, Gas , Drug Contamination , Ethanol , Naphthalenes , Resins, Synthetic , Chemistry , Sodium Hydroxide , Spectrophotometry, Ultraviolet , Toluene
20.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1103-1105, 2004.
Article in Chinese | WPRIM | ID: wpr-284540

ABSTRACT

<p><b>OBJECTIVE</b>To study the anti-asthmatic effect of sodium ferulate (SF) and its mechanism in guinea pig asthmatic model.</p><p><b>METHODS</b>Guinea pigs were sensitized with ovalbumin as animal asthmatic model and treated with 3 different concentration of SF for 8 days. Levels of endothelin (ET) and nitric oxide (NO) in blood and lung tissue, and eosinophil (EOS) in blood and bronchoalveolar lavage fluid (BLAF) was counted at the end of trial.</p><p><b>RESULTS</b>SF could significantly lower the ET content and increase the NO concentration in serum and lung tissue, reduce the number of EOS in blood and BLAF (P < 0.05, P < 0.01). Stronger effect was showed in the high dose group.</p><p><b>CONCLUSION</b>Mechanism of anti-asthmatic action of SF might be to increase NO concentration, lower ET content, alleviate EOS infiltration.</p>


Subject(s)
Animals , Female , Male , Anti-Asthmatic Agents , Pharmacology , Asthma , Blood , Bronchoalveolar Lavage Fluid , Cell Biology , Coumaric Acids , Pharmacology , Endothelin-1 , Blood , Eosinophils , Pathology , Guinea Pigs , Nitric Oxide , Blood , Ovalbumin
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