CAJ | 학술논문

目的探讨组胺受体4种亚型(H1、H2、H3、H4受体)在小儿尿道横纹肌中和小鼠成肌干细胞C2C12分化过程中的表达.方法收集临床小儿尿道横纹肌标本,石蜡包埋,组织切片,免疫组织化学染色检测Ⅲ、H2、H3、H4受体.诱导小鼠成肌干细胞C2C12成肌分化,实时定量聚合酶链反应测量成肌分化早期标志物desmin,中期标志物myogenin,晚期标志物肌球蛋白重链和组胺4种亚型受体mRNA的表达.免疫荧光染色检测各分化标志物和组胺H3受体蛋白质的表达情况.结果成肌分化过程中desmin mRNA在分化2、4、6d后的表达量分别为未分化肌母细胞的12、68、60倍;myogenin mRNA在分化2、4、6d后的表达量分别为肌母细胞的631、1 408、914倍;肌球蛋白重链mRNA的表达在分化2、4、6d后,表达量分别为未分化的肌母细胞的7 718、9448、286 288倍.H1受体mRNA在分化6d后,表达量约为未分化肌母细胞的25％;H2受体mRNA在未分化肌母细胞与分化各阶段的细胞中的表达均无明显变化(F=1.47,P＞0.05);H3受体mRNA在分化2、4、6d后的表达量是未分化肌母细胞的28、103、198倍;未检测到组胺受体H4受体mRNA的表达.免疫荧光染色发现H3受体蛋白染色随着分化荧光染色强度增加.小儿尿道横纹肌的免疫组织化学染色发现H1、H2、H3受体染色阳性,其中H1受体阳性程度最强,H3受体中度阳性,H2受体弱阳性.结论小儿尿道横纹肌中和小鼠成肌干细胞分化过程中发现组胺亚型H1、H2、H3受体的表达,H3受体的表达量随着成肌分化过程而增加,预示着其可能在成熟横纹肌细胞中发挥功能.
목적 탐토조알수체4충아형(H1、H2、H3、H4수체)재소인뇨도횡문기중화소서성기간세포C2C12분화과정중적표체.방법 수집림상소인뇨도횡문기표본,석사포매,조직절편,면역조직화학염색검측Ⅲ、H2、H3、H4수체.유도소서성기간세포C2C12성기분화,실시정량취합매련반응측량성기분화조기표지물desmin,중기표지물myogenin,만기표지물기구단백중련화조알4충아형수체mRNA적표체.면역형광염색검측각분화표지물화조알H3수체단백질적표체정황.결과 성기분화과정중desmin mRNA재분화2、4、6d후적표체량분별위미분화기모세포적12、68、60배;myogenin mRNA재분화2、4、6d후적표체량분별위기모세포적631、1 408、914배;기구단백중련mRNA적표체재분화2、4、6d후,표체량분별위미분화적기모세포적7 718、9448、286 288배.H1수체mRNA재분화6d후,표체량약위미분화기모세포적25％;H2수체mRNA재미분화기모세포여분화각계단적세포중적표체균무명현변화(F=1.47,P＞0.05);H3수체mRNA재분화2、4、6d후적표체량시미분화기모세포적28、103、198배;미검측도조알수체H4수체mRNA적표체.면역형광염색발현H3수체단백염색수착분화형광염색강도증가.소인뇨도횡문기적면역조직화학염색발현H1、H2、H3수체염색양성,기중H1수체양성정도최강,H3수체중도양성,H2수체약양성.결론 소인뇨도횡문기중화소서성기간세포분화과정중발현조알아형H1、H2、H3수체적표체,H3수체적표체량수착성기분화과정이증가,예시착기가능재성숙횡문기세포중발휘공능.
Objective To explore the expressions of histamine receptor subtypes (H1, H2, H3, H4 receptor) in children's mid-urethral striated muscles and during mouse C2C12 striated myogenesis.Methods Children's mid-urethral striated muscle samples were paraffin embedded and tissue sections were made, then immunohistochemical staining was used to check H1, H2, H3, H4 receptors.C2C12 myogenesis was induced, the early differentiation early markers of desmin, middle marker of myogenin, late marker of myosin heavy chain and histamine 4 receptor subtype mRNA were checked by quantitative real-time polymerase chain reaction.Immunofluorescence staining was done to check 3 differentiation markers and histamine H3 receptor protein.Results During myogenesis, the expression of desmin mRNA in the differentiation of 2,4,6 days were 12,68,60 times as many as that of the undifferentiated myoblasts;the expression of myogenin mRNA in the differentiation of 2,4,6 days were 631,1 408,914 times as many as that of the undifferentiated myoblasts;the expression of myosin heavy chain mRNA in the differentiation of 2,4,6 days were 7 718,9 448,286 288 times as many as that of the undifferentiated myoblasts.The expression level of H1 receptor mRNA in the differentiation of 6 days was about 25％ to undifferentiated cells;the expression of H2 receptor mRNA in undifferentiated cells and differentiated cells groups had no significant difference (F =1.47, P ＞ 0.05);the expression of H3 receptor mRNA in the differentiation of 2,4,6 days was 28,103,198 times to undifferentiated cells;H4 receptor mRNA was not detected.In immunofluorescence staining, H3 receptor protein staining intensity increased with the differentiation.Immunohistochemistry of pediatric urethral striated staining indicated that H1, H2, H3 receptor staining was positive,H1 receptor showed strong positive staining, H3 receptor moderate positive staining,and H2 receptor showed weak positive staining.Conclusions Histamine receptor subtypes of H1 receptor, H2 receptor and H3 receptor were found during mouse striated myogenesis and in the children's mid-urethral striated muscles.The increasing expression of H3R with myogenesis might indicate it plays a role in mature striated muscle cells.