CAJ | 학술논문

目的：研究膦甲酸钠（ PFA）对高磷诱导的大鼠血管平滑肌细胞（ VSMCs ）钙化的影响及可能的作用机制。方法体外原代培养大鼠胸主动脉VSMCs，经形态学和免疫细胞化学鉴定后，分别加入10 mmol／L的β-甘油磷酸（ HP组）和10 mmoL／L HP＋1 mol／L PFA （ PFA组），另设空白对照组。测定各组细胞钙含量，并分别用RT-PCR和Western blotting法检测VSMCs 核心结合因子α1（Cbfα1）mRNA及蛋白在不同时间点的动态表达情况。结果 HP组和PFA组细胞钙含量均高于空白对照组（ P均＜0．05），且PFA组细胞钙含量较HP组明显降低（ P＜0．05）。 HP组Cbfα1 mRNA和蛋白的表达呈“峰-谷-峰”样（P＜0．05），且第一峰值高于第二峰值（P＜0．05）；PFA组Cbfα1 mRNA表现为第二高峰前移，分别于6 h及1 d达峰（P均＜0．05），而蛋白水平达峰时间延后，分别于12 h和4 d达峰（P＜0．05），且第二峰值均低于第一峰值（P＜0．05）。结论 PFA能抑制高磷诱导的大鼠VSMCs钙化，其可能是通过影响高磷诱导的VSMCs表型转化来实现。
목적：연구련갑산납（ PFA）대고린유도적대서혈관평활기세포（ VSMCs ）개화적영향급가능적작용궤제。방법체외원대배양대서흉주동맥VSMCs，경형태학화면역세포화학감정후，분별가입10 mmol／L적β-감유린산（ HP조）화10 mmoL／L HP＋1 mol／L PFA （ PFA조），령설공백대조조。측정각조세포개함량，병분별용RT-PCR화Western blotting법검측VSMCs 핵심결합인자α1（Cbfα1）mRNA급단백재불동시간점적동태표체정황。결과 HP조화PFA조세포개함량균고우공백대조조（ P균＜0．05），차PFA조세포개함량교HP조명현강저（ P＜0．05）。 HP조Cbfα1 mRNA화단백적표체정“봉-곡-봉”양（P＜0．05），차제일봉치고우제이봉치（P＜0．05）；PFA조Cbfα1 mRNA표현위제이고봉전이，분별우6 h급1 d체봉（P균＜0．05），이단백수평체봉시간연후，분별우12 h화4 d체봉（P＜0．05），차제이봉치균저우제일봉치（P＜0．05）。결론 PFA능억제고린유도적대서VSMCs개화，기가능시통과영향고린유도적VSMCs표형전화래실현。
Objective To explore the effect and mechanism of phosphonoformic acid ( PFA) on β-glycerophosphate-incudced calcification in rat vascular smooth muscle cells (VSMCs).Methods Primary VSMCs obtained from rat thoracic aorta in vitro were cultured .After being identified with morphological and immunocytochemistry , VSMCs were added with 10 mmol/Lβ-glycerophosphate (HP group) and 10 mmol/L HP +1 mmol/L PFA (PFA group), meanwhile, the control group was set .RT-PCR and Western blot were used to detect the expression of mRNA and protein of core binding factor α1 (Cbfα1) at different time points, respectively.Results Compared with the control group, the cell calcification in the HP group and PFA group was increased (all P<0.05), while compared with HP group, the calcium deposition was signifi-cantly reduced in the PFA group(P<0.05).In the HP group, the expression of mRNA and protein of Cbfα1 were in a“peak-valley-peak” trend (P<0.05), and the first peak was higher than the second one (P<0.05).In the PFA group, the second peak of Cbfα1 mRNA moved forward, which reached the top at 6h and 1d respectively(all P<0.05).Unlike the expression of mRNA, the protein of Cbfα1 reached the top at 12h and 4d, respectively(P<0.05), and the second peak was lower than the first one(P<0.05).Conclusion PFA inhibitsβ-glycerophosphate-incudced VSMCs calcification possibly by affecting the hyperphosphate-induced phenotypic transdifferentiation of VSMCs .