摘要
Purpose:To investigate the effect of apigenin on gap junctional intercellular communication (GJIC) in human Tenon's capsule fibroblasts (HTFs) and its underlying mechanism. Methods:After a 48 h treatment of cultured HTFs with apigenin.(80 μmol/L),the GJIC was detected by a scrape-loading/dye transfer technique with Lucifer yellow dye and rhodamine (Rh) dextran. The coupling index represents a quantification of GJIC where a high coupling index is associated with a greater number of cells demonstrating cell-cell communication through gap junction channels.The changes in connexin 43 (Cx43) distribution and the expression of Cx43 at the protein and mRNA levels were statistically compared between the two groups by means of immunocytochemistry, western blotting,and real-time polymerase chain reaction (PCR). Results:The functioning of GJIC in the HTFs was significantly enhanced after 48 hours by apigenin treatment when compared with the control cells. In the apigenin group, the intercellular dye transfer grade was above 9, while this value was only grade 3-4 in the control group. The coupling index was significantly increased up to 9.205±0.3621 in the apigenin group,compared with 5.1775 ±0.3177 in the control group (F=279.581, P=0.000). The expression of Cx43 at the protein and mRNA levels was significantly up-regulated in the apigenin group compared with the control group. Conclusion:Apigenin can significantly enhance the function of GJIC in HTFs by up-regulating the expression of Cx43 at both the protein and mRNA levels,suggesting that the enhancement of GJIC in HTFs by apigenin probably acts as an important mechanism underlying the inhibitory effect of apigenin on HTF proliferation.
Purpose： To investigate the effect of apigenin on gap junc- tional intercellular communication （GJIC） in human Tenon＇s capsule fibroblasts （HTFs） and its underlying mechanism. Methods： After a 48 h treatment of cultured HTFs with api- genin （80 umol/L）, the GJIC was detected by a scrape-load- ing/dye transfer technique with Lucifer yellow dye and rho- damine （Rh） dextran. The coupling index represents a quan- tification of GJIC where a high coupling index is associated with a greater number of cells demonstrating cell-cell commu- nication through gap junction channels. The changes in con- nexin 43 （Cx43） distribution and the expression of Cx43 at the protein and mRNA levels were statistically compared be- tween the two groups by means of immunocytochemistry, western blotting, and real-time polymerase chain reaction （PCR）. Results： The functioning of GJIC in the HTFs was signifi- cantly enhanced after 48 hours by apigenin treatment when compared with the control cells. In the apigenin group, the intercellular dye transfer grade was above 9, while this value was only grade 3-4 in the control group. The coupling index was significantly increased up to 9.205±0.3621 in the apigenin group, compared with 5.1775±0.3177 in the control group （F=279.581, P=0.000）. The expression of Cx43 at the protein and mRNA levels was significantly up-regulated in the api- genin group compared with the control group. Conclusion： Apigenin can significantly enhance the function of GJIC in HTFs by up-regulating the expression of Cx43 at both the protein and mRNA levels, suggesting that the en- hancement of GJIC in HTFs by apigenin probably acts as an important mechanism underlying the inhibitory effect of api- genin on HTF proliferation.
基金
supported by Shandong Provincial Natural Science Foundation Project (No.ZR2010HM015)
