补肾活血固齿方对MC3T3-E1细胞超微结构及Ⅰ型胶原蛋白影响的实验研究  

Regulatory effects of the Bushen Huoxue Guchi Recipe on the ultrastructure of MC3T3-E1 cells and the mRNA level of collagen typeⅠ

作　　者：杨梦华[1] 杨运田[2] 都广艳[3] 赵玲 高毅[1] 王双双[1] 李蓓[1] YANG Menghua;YANG Yuntian;DU Guangyan;ZHAO Ling;GAO Yi;WANG Shuangshuang;LI Bei(Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 05001)

机构地区：[1]河北省中医院口腔科,石家庄050011 [2]河北医科大学口腔医院综合科 [3]石家庄医学高等专科学校 [4]北京市通州区妇幼保健院口腔科

出　　处：《现代口腔医学杂志》2023年第5期289-294,共6页Journal of Modern Stomatology

基　　金：河北省中医药管理局科研计划项目(2022049)。

摘　　要：目的观察补肾活血固齿方对小鼠颅顶前成骨细胞(MC3T3-E1细胞)超微结构、Ⅰ型胶原蛋白(Collagen TypeⅠ,ColⅠ)mRNA的影响,探讨该方剂成骨的作用机理。方法MC3T3-E1细胞分别于补肾活血固齿方含药血清、无药血清、胎牛血清中培养,透射电镜分别于1 d、4 d、7 d、14 d,观察MC3T3-E1细胞的超微结构;同时应用QRT-PCR法检测培养1 d、2 d、3 d后,补肾活血固齿方对MC3T3-E1细胞ColⅠmRNA表达的影响。结果于含药血清培养的MC3T3-E1细胞,随着时间的延长细胞质中细胞器逐渐丰富,14 d时MC3T3-E1细胞呈成骨细胞样表型;无药血清组、胎牛血清组细胞的细胞器与含药血清组细胞相比较不丰富,14 d时MC3T3-E1细胞表面突起较少。1d、2 d、3 d时,无药血清组、胎牛血清组细胞的ColⅠmRNA表达均无明显差异(P>0.05);含药血清组细胞ColⅠⅠmRNA表达与其他2组相比明显增多(P<0.05)。结论补肾活血固齿方可能通过增加ColⅠmRNA的表达,促进MC3T3-E1细胞向成骨细胞样表型发展,从而促进骨的形成。Objective To observe the regulatory effects of the Bushen Huoxue Guchi Recipe on the ultrastructure of the mouse preosteoblast cell line MC3T3-E1 and the mRNA level of collagen type Ⅰ(ColⅠ),and to explore the mechanism of this traditional Chinese medicine(TCM)preparation in stimulating osteogenesis.Methods MC3T3-E1 cells were cultured in fresh medium containing rat serum and Bushen Huoxue Guchi Recipe(group A),rat serum medium free of Bushen Huoxue Guchi Recipe(group B),and fresh medium containing fetal bovine serum(FBS)(group C)for 14 days,respectively.On day 1,4,7 and 14,the ultramicroscopic changes of MC3T3-E1 cells were observed by transmission electron microscopy(TEM).On day 1,2 and 3,the mRNA level of ColⅠwas detected by qRT-PCR.Results Cytoplasmic organelles were gradually enriched in MC3T3-E1 cells of group A,showing an osteoblast-like phenotype on day 14.On the contrary,cytoplasmic organelles in MC3T3-E1 cells of group B and C were less abundant,and less cell protrusions were observed on day 14.There was no significant difference in the mRNA level of Col I in MC3T3-E1 cells of group B and C on day 1,2 and 3(P>0.05),which was significantly upregulated in those of group A(P>0.05).Conclusion Bushen Huoxue Guchi Recipe stimulates osteoblast differentiation and maturation in MC3T3-E1 cells and promotes bone formation by upregulating the mRNA level of ColⅠ.

关 键 词：补肾活血固齿方 牙周炎 MC3T3-E1细胞 超微结构 Ⅰ型胶原蛋白 

分 类 号：R781.4[医药卫生—口腔医学]