摘要
目的:通过研究接骨1号方含药血清对MC3T3-E1细胞NF-κb信号通路的影响,探讨接骨1号方促进骨折愈合的可能机制。方法:分别以接骨1号方高、中、低剂量、阳性对照药补佳乐和生理盐水予大鼠灌胃以制备大鼠含药血清,共5组,分别为JG1-H组、JG1-M组、JG1-L组、Est组、Control组,对MC3T3-E1细胞进行干预,7 d后,观察细胞形态,并采用RealTime RT-PCR法检测骨保护素(osteoprotegerin,OPG)、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)和核因子κb(Nuclear factorκb,NF-κb)mRNA表达,用Western blot法检测OPG、TNF-α和NF-κb蛋白的表达。结果:JG1-M组与Est组MC3T3-E1细胞OPG mRNA和蛋白表达水平均显著高于Control组(P<0.01)。JG1-M组MC3T3-E1细胞TNF-α和NF-κb mRNA和蛋白的表达均水平低于Control组(P<0.01)。结论:接骨1号方含药血清可能通过调控NF-κb信号通路促进MC3T3-E1细胞的成骨分化;接骨1号方可能通过抗炎促进骨折愈合。
Objective:To investigate the effect of serum containing Jiegu No.1 recipe on the NF-κb signal pathway of MC3 T3-E1 cell,and to explore the possible mechanism of promoting fracture healing by Jiegu No.1 recipe.Methods:The rat serum containing the drug was prepared by intragastric administration of Jiegu No.1 recipe with high,medium,low doses,Progynova and normal saline(NS).There were 5 groups:JG1-H group,JG1-M group,JG1-L group,Est group,and control group.After intervening in the MC3 T3-E1 cells,after 7 d,the cell morphology was observed,and the mRNA expression levels of osteoprotegerin(OPG),tumor necrosis factorα(TNF-α)and Nuclear factorκb(NF-κb)were detected by RealTime RT-PCR.The expression levels of OPG,TNF-αand NF-κb were measured by western blot.Results:The expression levels of OPG mRNA and protein in MC3 T3-E1 cells of JG1-M group and Est group were significantly higher than those of control group(P<0.01).The expressions levels of TNF-αand NF-κb mRNA and protein in MC3 T3-E1 cells of JG1-M group were lower than those of control group(P<0.01).Conclusion:The serum of Jiegu No.1 recipe may promote the osteogenic differentiation of MC3 T3-E1 cells by regulating NF-κb signaling pathway,and Jiegu No.1 recipe may promote fracture healing through anti-inflammatory.
作者
孙广江
姚啸生
杨鸫祥
SUN Guang-jiang;YAO Xiao-sheng;YANG Dong-xiang(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang Liaoning 110032,China)
出处
《中医药导报》
2019年第14期39-42,59,共5页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
国家中医药管理局项目(JDZX2015029)
