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ABSTRACT:Objective To explore the role of HIF‐1αin the pathogenesis of hepatopulmonary syndrome (HPS) and its relationship with GRP78 .Methods The HPS model in rats was induced by multiple pathogenic factor .The samples were assessed by using Western blotting analysis for HIF‐lα, GRP78 and VEGF164 . The expressions of VEGFR‐2 and CD105 were observed by using immunohistochemical staining .Results The protein level of HIF‐1αwas significantly increased in HPS group at week 8 compared with that at week 4 and 6 groups and corresponding normal control groups .With the development of HPS ,protein level of GRP78 was gradually increased at each time point significantly and reached the highest level at week 8 ;protein level of VEGF164 showed a similar change with GRP78 ,but the peak was at week 6 .Immunohistochemical results showed that the protein expressions of VEGFR‐2 and CD105 were gradually increased in lung tissue as HPS progressed .The protein level of GRP78 was positively correlated with HIF‐1α,VEGF164 ,VEGFR‐2 and CD105 ,respectively (P<0 .05) .Conclusion HIF‐1αis most likely together with GRP78 to play a critical role in promoting pulmonary microvascular remodeling in the pathogenesis of HPS in rats .
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OBJECTIVE@#To observe the influence of ropivacaine on the proliferation and migration of rat bone marrow mesenchymal stem cells (BMSCs) and provide basis for the clinical application of BMSCs.@*METHODS@#Rat BMSCs were isolated and cultured by adherence method. Surface markers of BMSCs were examined by flow cytometry. Multipotent differentiation of BMSCs was detected by induced adipogenesis, osteogenesis and muscular differentiation. Proliferation of BMSCs was examined by CCK-8 and Brdu incorporation after ropivacaine treatment at different concentrations. Migration of BMSCs was tested by cell scratch assay and Millicell experiment.@*RESULTS@#Cultured cells had representative appearance and surface markers of BMSC, and they had potential multiple differentiation. Ropivacaine treatment at 50 and 100 μmol/L significantly reduced the proliferation rate of BMSCs and Brdu incorporation rate. There was significant difference compared with the control group (P<0.05). Cellular scratch assay and migration experiment indicated that ropivacaine significantly reduced the migration of BMSCs. There was significant difference compared with the control group (P<0.05). All these mentioned effects of ropivacaine on BMSCs were dose-dependent. There was significant difference between groups (P<0.05).@*CONCLUSION@#Ropivacaine can significantly reduce the proliferation and migration of rat BMSCs, suggesting that the influence of local anesthetics on BMSCs has to be taken into account when BMSCs are used in clinical practice.
Subject(s)
Animals , Rats , Amides , Pharmacology , Bone Marrow Cells , Cell Differentiation , Cell Movement , Cell Proliferation , Cells, Cultured , Flow Cytometry , Mesenchymal Stem Cells , Cell Biology , RopivacaineABSTRACT
AIM: To observe the influence of lentiviral vectors expressing siRNA for survivin gene knockdown in A549 cells,sequentially as tools to explore the molecule pathogenesis and new gene therapy of lung adenocarcinoma.METHODS: The lentiviral vectors,which express survivin siRNA,were constructed and transfected into A549 cell strain.The titers of the lentiviruses were determined by 293T cells.The expressions of survivin and caspase-3 were detected by Western blotting and RT-PCR.The cell cycle and cell growth of A549 cells were examined by MTT and FCM.RESULTS: The expression of survivin was suppressed effectively by siRNA targeting survivin.The expression of survivin mRNA decreased by 97%.The expression of survivin protein decreased by 94%.The rate of cell growth was decreased.The G1 phase cells were increased,whereas S phase cells were decreased.CONCLUSION: The lentivirus vectors expressing siRNA for survivin can significantly inhibit gene expression and the cell growth,and markedly induce the apoptosis.It is hopeful to be a new gene therapy of lung adenocarcinoma.