CAJ | 학술논문

目的：探讨组胺H3受体（H3R）在小鼠成肌细胞C2C12成肌分化过程及分化后的横纹肌细胞中的表达和可能发挥的作用。方法：诱导C2C12细胞成肌分化，测量H3R和分化晚期标志物肌球蛋白重链mRNA和蛋白的表达；分化过程中加入H3R拮抗剂ciproxifan，测量分化早期标志物desmin、中期标志物myogenin和肌球蛋白重链mRNA的表达。 Fluo－4结合剂标记分化后的横纹肌胞内钙离子，测量双极交流电200 mA刺激下，H3R激动剂甲基组胺（RMeHA）对胞浆中钙离子浓度的影响。结果：H3R和肌球蛋白重链在成肌分化过程中表达量逐渐增加。 Ciproxifan在成肌分化过程中对3种分化标志物mRNA的表达与对照组相比无差异（ P＞0．05）。 RMeHA在浓度10 nmol／L～100μmol／L刺激细胞5～20 min，可呈钟形降低因交流电引起的肌浆钙离子浓度的升高（ P＜0．05），其中RMeHA 100 nmol／L在10 min和20 min对电刺激细胞中Ca2＋的抑制百分率最高。相同浓度的RMeHA在20 min和10 min时对Ca2＋的抑制率比其在5 min时高（P＜0．05）。结论：H3R可能在成肌分化过程中的作用不大，而在分化成熟细胞中可以降低电刺激引起的胞浆钙离子浓度的升高。
목적：탐토조알H3수체（H3R）재소서성기세포C2C12성기분화과정급분화후적횡문기세포중적표체화가능발휘적작용。방법：유도C2C12세포성기분화，측량H3R화분화만기표지물기구단백중련mRNA화단백적표체；분화과정중가입H3R길항제ciproxifan，측량분화조기표지물desmin、중기표지물myogenin화기구단백중련mRNA적표체。 Fluo－4결합제표기분화후적횡문기포내개리자，측량쌍겁교류전200 mA자격하，H3R격동제갑기조알（RMeHA）대포장중개리자농도적영향。결과：H3R화기구단백중련재성기분화과정중표체량축점증가。 Ciproxifan재성기분화과정중대3충분화표지물mRNA적표체여대조조상비무차이（ P＞0．05）。 RMeHA재농도10 nmol／L～100μmol／L자격세포5～20 min，가정종형강저인교류전인기적기장개리자농도적승고（ P＜0．05），기중RMeHA 100 nmol／L재10 min화20 min대전자격세포중Ca2＋적억제백분솔최고。상동농도적RMeHA재20 min화10 min시대Ca2＋적억제솔비기재5 min시고（P＜0．05）。결론：H3R가능재성기분화과정중적작용불대，이재분화성숙세포중가이강저전자격인기적포장개리자농도적승고。
AIM:To explore the expression and possible function of histamine H3 receptor (H3R) in striated myogenesis and the differentiated C2C12 cells.METHODS: H3R and myogenesis late marker myosin heavy chain (MHC) were detected at mRNA and protein levels during C2C12 myogenesis.H3R antagonist ciproxifan was added and the expression of the myogenesis early marker desmin, intermediate markers myogenin and MHC was detected.Differentia-ted myoblasts were loaded with Fluo-4 calcium indicator dye and the effect of R-( a)-methylhistamine ( RMeHA) on the cy-toplasmic calcium concentration was determined under the 200 mA electrical stimulation.RESULTS: The expression of H3R and MHC was increased during myogenesis.Ciproxifan incubation had no influence on the 3 striated myogenesis mar-kers (P>0.05).In C2C12 myoblasts, RMeHA (10 nmol/L~100 μmol/L) effectively diminished cytoplasmic calcium peak when the cells were electrically paced (P<0.05).The best inhibitory effect of RMeHA was observed at dose of 100 nM for 10 min and 20 min, which was higher than that for 5 min (P<0.05).CONCLUSION: H3R might have little effect on the myogenic differentiation, but diminishes cytoplasmic calcium peak of the differentiated myoblasts under electri-cal stimulation.