三磷酸腺苷敏感性钾离子通道在外源性硫化氢抑制r高糖诱导的成骨细胞损伤中的作用

摘要

目的 研究三磷酸腺苷敏感性钾离子通道(KATP)在硫化氢(H2S)抑制高糖(HG)诱导的成骨细胞损伤中的作用.方法 原代培养大鼠下颌骨成骨细胞并鉴定,将成骨细胞给予HG、H2S、KATP通道开放剂吡拉地尔(Pia)、阻断剂格列本脲(Gli)处理后,用Western blot检测KATP通道蛋白的表达水平,同时采用CCK8、实时逆转录聚合酶链反应(RT-PCR)、茜素红S染色分析方法检测其对成骨细胞增殖、分化及矿化的影响.结果 细胞KATP通道蛋白的表达水平在HG的影响下明显降低,H2S预处理明显抑制HG对KATP通道蛋白的下调作用和对成骨细胞增殖的影响,并防止HG引起的成骨细胞分化和矿化的减少;相反,KATP通道阻断剂能明显阻断H2S对成骨细胞的上述保护作用.结论 H2S通过KATP通道抑制了HG诱导的成骨细胞损伤.%Objective The aim of this study is to identify the role of adenosine triphosphate-sensitive potassium channel (KATP) in hydrogen sulfide (H2S)-induced inhibition of high glucose (HG)-induced osteoblast damage.Methods Osteoblasts from rat mandible were cultured and identified. The osteoblasts were then treated with HG, H2S, KATP channel opener pinacidil (Pia), and KATP channel blocker glibenclamide (Gli). Western blot method was performed to detect the expression of KATP channel protein. CCK8, reverse transcriptase polymerase chain reaction (RT-PCR) , and image analysis were used to determine the effects of H2S-KATP on the proliferation, differentiation, and mineralization of osteoblasts.Results The expression of KATP channel protein in osteoblasts was significantly decreased under the influence of HG. H2S pretreatment significantly inhibited HG on KATP channel protein down-regulation. Moreover, H2S pretreatment significantly inhibited the effect of HG on the proliferation of osteoblasts, thereby preventing HG-induced inhibition of osteoblasts differentiation and mineralization. Meanwhile, the KATP channel blocker effectively blocked the H2S on osteoblasts and had a pro-tective effect.Conclusion Through the KATP channel, H2S inhibited osteoblasts damage induced by HG.