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ObjectiveTo systematically analyze the chemical components of QiLing Wenshen （QLWS） formula and explore the key active components and mechanism of the formula in the treatment of polycystic ovary syndrome （PCOS）. MethodThe chemical components of QLWS formula were systematically identified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF/MSE） combined with comparison with reference substances， literature data， and databases. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and SwissADME were employed to screen the active components for network pharmacological analysis. SwissTargetPrediction， GeneCards， DisGeNET， and DrugBank were used to obtain the potential components and targets of the formula for the treatment of PCOS. The protein-protein interaction （PPI） network was constructed via STRING database for further screening of the core targets. Gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of core targets were carried out with DAVID database. Molecular docking was performed in MOE 2019. ResultA total of 90 components of QLWS formula were identified， and 32 active components and 45 core targets for treating PCOS were obtained. GO annotation obtained 429 terms and KEGG pathway enrichment screened out 110 signaling pathways， mainly involving phosphatidylin-ositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， estrogen signaling pathway， and hypoxia inducible factor-1 （HIF-1） signaling pathway. The molecular docking revealed that key active components in QLWS formula were icariin， salvianolic acid A\B\C， wogonin， magnoflorine， etc.， which may play a role in treating PCOS through regulating mitogen-activated protein kinase 1 （MAPK1）， epidermal growth factor receptor （EGFR）， mitogen-activated protein kinase 3 （MAPK3）， etc. ConclusionThis study preliminarily predicted that several key active components of QLWS formula could treat PCOS via multiple targets and multiple pathways based on UPLC-Q-TOF/MSE and network pharmacology， which could provide ideas and references for the study of pharmacodynamic material basis and mechanism of action of the formula.
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For SARS-CoV-2-induced pneumonia, “Diagnosis and Treatment of Novel Coronavirus Pneumonia (trial version 3)” released by the National Health Commission of the People’s Republic of China and the National Administration of Traditional Chinese Medicine on January 22, 2020, first proposed the traditional Chinese medicine (TCM) treatment plan, which was gradually updated to the seventh edition. Patients diagnosed with severe COVID-19 often developed dyspnea and/or hypoxemia one week after onset, requiring respiratory support and circulation support. Through literature research, it was found that Chinese materia medica (CMM) with tonifying qi and activating blood effect could effectively improve hypoxemia of pneumonia-related diseases. This article summarizes the mechanism and clinical application of tonifying qi and activating blood CMM in the treatment of novel coronavirus pneumonia-associated hypoxemia, and provides a reference for clinical treatment of COVID-19.
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<p><b>OBJECTIVE</b>To investigate the effect of adenoviral-mediated exogenous HGF (Ad-HGF) gene transfer on lung angiogenesis in the rabbit lung in rabbits with hyperkinetic pulmonary artery hypertension.</p><p><b>METHODS</b>A thoracotomy was performed through a midsternal incision in 1-month-old immature rabbit and an anastomosis between the left innominate artery and the pulmonary trunk was made to establish a chronic patent left to right shunt. Three months later, animals were randomly assigned to receive either Ad-HGF (2 x 10(9) Pfu in 0.2 ml PBS, H1 group), repeated administration of Ad-HGF after one week (H 2 group), Ad-GFP (2 x 10(9) Pfu in 0.2 ml PBS, G group), or PBS (0.2 ml, C group) by the intratracheal method of gene transfection. After two weeks, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical examination was performed to identify HGF mRNA and HGF protein expression. The capillary density and small pulmonary artery density were determined by immunostained with antibodies against factor VIII and alpha-SMA. After 1 month, the collateral vessels were evaluated by angiogram under digital subtraction angiography (DSA).</p><p><b>RESULTS</b>Two weeks after Ad-HGF transfection, 484 bp bands could be found by RT-PCR in H1 and H2 groups, but not in other groups. The expression of HGF protein could be detected on alveolar epithelium and pulmonary vessel endothelium by immunohistochemistry examination. The number of factor VIII-positive pulmonary capillaries was also significantly increased in the H1 and H2 groups compared with the C and G groups (P < 0.05). The capillary density reached (17.0 +/- 3.3) mm(2) and (19.7 +/- 2.8) mm(2) in the H1 and H2 group, respectively, whereas it remained (13.2 +/- 3.2) mm(2) in the C group and (13.5 +/- 2.4) mm(2) in the G group (P < 0.05). One month after Ad-HGF transfection, the number of small pulmonary arteries was significantly increased in H1 and H2 group compared with control groups (P < 0.05). The collateral vessels were more abundant in HGF transfection groups than that in the two control groups reviewed by angiogram under digital subtraction angiography (DSA).</p><p><b>CONCLUSION</b>In vivo gene transfection with HGF by means of the intra-tracheal injection could induce pulmonary angiogenesis in the early stage and small pulmonary arterial angiogenesis in later stage.</p>
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Animals , Female , Male , Rabbits , Disease Models, Animal , Hepatocyte Growth Factor , Genetics , Hypertension, Pulmonary , Lung , Neovascularization, Physiologic , TransfectionABSTRACT
Transfer of vascular endothelial growth factor (VEGF) gene to ischemic myocardium may provide a useful approach for angiogenesis and improve cardiac performance. However, uncontrolled expression of VEGF in vivo may result in certain side effects, such as hemangioma formation, retinopathy, and tumor development. We investigated the feasibility of using the nine copies of hypoxic response element (HRE) to control the expression of human VEGF(165) (h-VEGF(165)) under anoxic condition at cell level and also observed the synchron of h-VEGF(165) mRNA and protein expressions. Recombinant adeno-associated viral (rAAV) vector was prepared by using the three-plasmid system and cotransfected to human embryo kidney 293T cells by the calcium phosphate precipitates method. The rAAV vector was purified by chloroform-PEG8000/NaCl-chloroform and added to cultured myocardiocytes. Myocardiocytes of Sprague-Dawley rat were cultured in serum-free medium and then randomly divided into eight groups. Group I: cultured under normoxic conditions (21% O2) for 8 h as control; Group II: cultured under anoxic conditions (1% O2) for 8 h; Group III: cultured under normoxic conditions (21% O2) for 8 h with gene transfer; Group IV: cultured under anoxic conditions (1% O2) for 8 h with gene transfer; Group V, VI, VII: cultured under anoxic conditions (1% O2) for 8 h with gene transfer and then tured to normoxic conditions (21% O2) for 4, 8 or 12 h, respectively; Group VIII: cultured under anoxic conditions (1% O2) for 20 h with gene transfer. After completion of cell culture, the amount of h-VEGF(165) protein in culture supernatant was quantified by using enzyme linked immunosorbent assay (ELISA). Expression of h-VEGF(165) protein in cultured cardiacmyocytes was also evaluated by immunofluorescence. RT-PCR was employed to detect the expression of h-VEGF(165) mRNA. The results revealed that there were no expressions of h-VEGF(165) mRNA and protein in groups I, II, III, VI and VII. After gene transfer, the expressions of h-VEGF(165) protein and mRNA were significantly higher in groups IV and VIII than those in other groups (P<0.01); Immunofluorescence positive cells were observed in groups IV, V and VIII. RT-PCR revealed that a 484-bp strip can be found in groups IV and VIII, but unavailable in other groups. We conclude that HRE is a promising regulator for h-VEGF(165) gene expression following the changes of oxygen environment. HRE can induce the expression of h-VEGF(165) gene after hypoxia, but in normal oxygen condition, the expression of h-VEGF(165) was inhibited. Although expression of h-VEGF(165) mRNA ceased in normal oxygen condition under the control of HRE, expression of h-VEGF(165) protein was hysteretic to h-VEGF(165) mRNA expression.