ABSTRACT
Objective:To deeply explore the potential mechanism of Kangmin Zhisou Granules in the treatment of bronchial asthma through network pharmacology method; To verify it with animal experiments.Methods:The active components and corresponding target information of Kangmin Zhisou Granules were screened with the help of BATMAN-TCM database, and the related disease targets of bronchial asthma were obtained through GeneCards and OMIM databases. The drug targets and bronchial asthma targets were intersected and imported String database was used to establish PPI network. Cytoscape 3.9.1 software was used to draw the network diagram of "Chinese materia medica-active components-intersection targets" and the core targets were screened. GO and KEGG enrichment analysis was performed on the core targets using DAVID database. A mouse model of asthma induced by ovalbumin was prepared. After the intervention of Kangmin Zhisou Granules, the pathological changes of mouse lung tissue were observed, and the contents of serum TNF-α, IL-6, IL-1 β were detected by ELISA.Results:Totally 240 active components and 1 364 potential targets were obtained from Kangmin Zhisou Granules. Tumor necrosis factor (TNF), interleukin-6 (IL-6), protein kinase B (AKT1), albumin (ALB), interleukin 1-beta (IL-1β) and other 11 core targets were obtained after screening. The results of GO enrichment analysis showed that the treatment of bronchial asthma by Kangmin Zhisou Granules mainly involved the positive regulation of protein phosphorylation, the regulation of inflammatory response, lipopolysaccharide response and other biological processes, as well as TNF, activated protein kinase (MAPK), interleukin-17 (IL-17) and other signaling pathways. Animal experiments confirmed that Kangmin Zhisou Granules could reduce the expression levels of TNF-α, IL-6 and IL-1β in serum ( P<0.05), and reduce the infiltration of inflammatory cells in the lung tissue of mice, thereby relieving asthma symptoms. Conclusion:Kangmin Zhisou Granules may exert anti-inflammatory effects by acting on TNF-α, IL-6, IL-1β and other targets to alleviate asthma symptoms.
ABSTRACT
On the basis of the origin comparison of known endothelial genesis inhibitors, a 417-bp cDNA fragment was amplified from umbilical cord by RT-PCR and cloned into the expression vector pPIC9, followed by transformation into Pichia pastoris GS115. The resulted yeast was induced with methanol to express recombinant protein. The resulted protein was purified from culture broth and designated as EDI-8t. The in vitro study showed that EDI-8t, originated from collagen VIII, could specifically inhibit the growth and migration of bovine aortic endothelial cells (BAEC) stimulated by basic fibroblast growth factor (bFGF). The protein also exhibited the activity to cause cell apoptosis. In vivo EDI-8t showed the identical activity comparing with endostatin to inhibit the growth of liver tumor transplanted into nude mice. Interestingly, EDI-8t showed higher activity than endostatin to inhibit tumor growth in metastatic model of melanoma mice.
Subject(s)
Animals , Cattle , Humans , Mice , Amino Acid Sequence , Angiogenesis Inhibitors , Genetics , Antineoplastic Agents , Pharmacology , Base Sequence , Cells, Cultured , Collagen Type VIII , Chemistry , Genetics , Endothelium, Vascular , Metabolism , Genetic Vectors , Genetics , Human Umbilical Vein Endothelial Cells , Chemistry , Mice, Nude , Molecular Sequence Data , Pichia , Genetics , Metabolism , Recombinant Proteins , Genetics , PharmacologyABSTRACT
BACKGROUND: Promoting immunological function of patients with chronic obstructive pulmonary disease (COPD) can control development of COPD.OBJECTIVE: To investigate the effect of bufei yishen granule on pulmonary ventilation function and immunological function of COPD patients and compare with placebo.DESIGN: A randomized grouping comparison and placebo controlled study.SETTING: Respiration Department of the First Affiliated Hospital of Henan Traditional Chinese Medical.PARTICIPANTS: A total of 62 COPD patients selected from Department of Respiration of the First Affiliated Hospital of Henan College of Traditional Chinese Medicine from January 1999 to October 2004,were randomly divided into two groups :Observation group and control group with 31 in each group.METHODS: Patients in observation group were treated with bufei yishen granule consisting of ginseng, mongolian milkvetch root, largehead atractylodes rhizome, divaricate saposhnikovia root, dwarf lilyturf tuber, Chinese magnoliavine fruit, malytea scurfpea fruit, medicinal evodia fruit, Chinese cster pillar fungus, Chinese eaglewood wood, scorpion, almond, thunberg fritillary bulb, szechwan lovge rhizome, etc., produced by Pharmaceutic Department of the First Affiliated Hospital of Henan College of Traditional Chinese Medicine. Each granule of 1 g contained an equivalent of 5.78 g raw drugs. Three times a day with 10 g for each; and patients in control group were treated with placebo consisting of hawthorn fruit and malt. The dosage and medication were the same as those in observation group. Both therapies were respectively administered in a treatment course of two months. Before and after the therapy course,the followed indexes were measured.① Assay of immunological function: The levels of IgG, IgA and IgM from blood serum were detected with simple agar diffusion.② Assay of Ag level in nucleolus organizer region (Ag-NORs): The ratios non-histone staining acidity non-histone vs core area (IS%) was calculated by staining the activating lymphocytes of the blood by silver staining technique with microscope image analysis technique. ③ Assay of pulmonary function: The pulmonary function was measured by Sensor Medics Ros System in all of the people with the levels of forced expiratory volume in first second (FEV1.0), mean maximu expiratory flow (MMEF), peak expiratory flow (PEF), once after and before treatment. The examination was repeated three times in order to obtain the peak number. The difference of the three examinations was within ±5%.MAIN OUTCOME MEASURES: Comparison of T lympholeukocyte subpopulation and pulmonary ventilation function in the patients before and after 1 course.RFSULTS: Data of totally 62 patients was entered the final analysis without any loss.① Comparison of pulmonary ventilation function of patients in the two groups before and after 1 course: FEV1.0, MMEF, PEF,and ratio of FEV1.0 and forced vital capacity (FEV1%) in observation group were significant higher than those before treatment (t=2.12-3.41,P ＜ 0.05-0.01), thoseindexes in observation group were higher than those in control group (t=2.54-3.17, P ＜ 0.05-0.01). ② Changes of T lymphocyte subgroups of patients in the two groups before and after 1 course:The levels of CD3+, CD4+, CD4+/CD8+ in observation group after 1 course were high.er than those before treatment (t=2.71-13.20, P ＜ 0.01), but the level of CD8+ was decreased as compared with that before treatment (t =8.63, P ＜ 0.01). The levels of CD3+, CD4+, CD4+/CD8+ in observation group were higher than those in control group (t=2.85-11.84, P ＜ 0.01), but the level of CD8+ was decreased as compared with that in control group (t =5.83, P ＜ 0.01).CONCLUSION: The bufei yishen granule can obviously improve the pulmonary ventilation function and immunological function of COPD patients,and its intervention is superior to that of placebo.
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Objective:To analyse the property of voltage-dependent potassium 〔K(v)〕 channel in healthy people′s peripheral lymphocyte so as to contribute the control for property alteration under some pathological condition,and to try to find the new subset of this channel.Methods:Patch-clamp whole cell recording technique was used.Results:In the recorded 39 cells,activated voltage of the channels was -40.3±2.5 Mv.No inactivation phenomenon appeared under repeated stimulation.The closing time of the channels was 116.3+8.2 ms under the repolarization,and the current could be inhibited by 10 mmol/L TEA.Conclusion:There might be only one type of K(v) channel in human peripheral blood lymphocytes,and its properties quite resumble the type of n K(v) channel in mice.