ABSTRACT
Aim To study the effects of L-borneol on the chloride channel and cell volume of human umbili-cal vein endothelial cells (HUVECs). Methods Whole-cell patch-clamp technique was used to record chloride currents. The expression of ClC-3 protein was down-regulated by siRNA interference technique. The cell volume was measured by dynamic image analysis. Results 20 nmol·L-1L-borneol significantly activa-ted chloride current in HUVEC (79.59 ± 4.90) pA/pF, which could be inhibited by chloride channel blockers,NPPB and DIDS. The outward current inhib-itory rate of NPPB was (95.57 ± 2.57)%, while that of DIDS was (97.28 ± 6.36)%. The chloride current activated by L-borneol significantly decreased after the silence of ClC-3 (27.03 ± 3.89) pA/pF. Cell volume was markedly reduced by L-borneol (14.38 ± 1.58)%,which was inhibited after NPPB appliance. Conclusion L-borneol can activate ClC-3 chloride channel in HUVECs, which induces Cl- outflow then cell volume decrease.
ABSTRACT
Aim To study the effect of oxymatrine (OMT)on protecting human umbilical vein endothelial cells (HUVECs)from toxicity induced by high glucose in vitro.Methods The HUVECs were cultured with medium containing different concentrations of glucose or OMT. The cells were randomly divided into 6 groups:5.5 mmol·L -1 control group(Control),22.2 mmol·L -1 high glucose (22.2 mmol·L -1 ),44.4 mmol·L -1 high glucose (44.4 mmol·L -1 ),control+OMT(control +OMT),22.2 mmol·L -1 high glu-cose +OMT(22.2 mmol·L -1 +OMT),44.4 mmol ·L -1 high glucose +OMT (44.4 mmol · L -1 +OMT).The protective effect of oxymatrine was as-sessed by MTS assays.The expression of A2B in HU-VECs was detected by Real-time quantitative PCR (qPCR)and Western Blot methods.Results The glucose was shown to have caused cytotoxicity in HU-VECs.Oxymatrine (3 μmol·L -1 )was found to have protected HUVECs from glucose toxicity effectively, and reduced the expression of A2B significantly.Con-clusion Oxymatrine can obviously protect HUVECs from cytotoxicity induced by high glucose and the effect is performed partly by decreasing A2B expression.
ABSTRACT
Objective To study the effects of Treponema pallidum membrane protein Tpp17 on ac-tivation of human umbilical vein endothelial cells (HUVECs) in vitro and to understand its role in the immu-nopathogenesis of syphilis .Methods HUVECs were co-cultured with recombinant protein Tpp 17.Then the expressions of TNF-α, MCP-1, ICAM-1 and E-selectin at mRNA and protein levels in supernatants were re-spectively detected by enzyme-linked immunosobent assay ( ELISA ) and fluorescent real-time quantitative PCR.The adhesive ability of THP-1 cells was observed by fluorescence microscopy after co-cultured pretrea-ted HUVECs with recombinant protein Tpp 17 with Calcein-AM labeled THP-1.Pretreated HUVECs were cultured in the lower chamber of Transwell with recombinant protein Tpp 17 and monocytic THP-1 cells were cultured in the upper chamber of Transwell .After that, the migration of monocytic THP-1 cells was evalua-ted by using fluorescence microscopy .Results Compared with the blank control group , the expression of TNF-α, ICAM-1, E-selectin, especially the MCP-1, were enhanced by recombinant protein Tpp 17 of Trepo-nema pallidum.Moreover, Tpp17 improved the adhesive ability and migration of monocytic THP-1 cells, es-pecially the latter.Conclusion Treponema pallidum membrane protein Tpp17 might play a certain role in the immunopathogenesis of syphilis by enhancing the expression of TNF-α, MCP-1, ICAM-1 and E-selectin, and by promoting the adherence ability and migration of monocytic THP-1 cells.