左归丸含药血清对BMSCs成骨分化过程中Cx43表达及GJIC功能的影响  

Effect of Zuogui Pill Medicated Serum on Expressions of Cx43 and GJIC Function During Osteogenic Differentiation of BMSCs

作　　者：桑红灵 孙志博[2] 陶春晖 章程鹏 赵敏 SANG Hongling;SUN Zhibo;TAO Chunhui;ZHANG Chengpeng;ZHAO Min(Hubei University of Chinese Medicine;Department of Orthopedics,Pu-Ai Hospital,Tongji Medical College,Huazhong University of Science and Technology; , Wuhan 430034, Hubei, Chin)

机构地区：[1]湖北中医药大学中医临床学院,湖北武汉430061 [2]华中科技大学同济医学院附属普爱医院骨科,湖北武汉430034

出　　处：《辽宁中医杂志》2018年第3期466-469,I0001,共5页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81403285,81501894);国家中医药管理局“周安方全国名老中医药专家传承工作室”建设项目([2014]20)

摘　　要：目的：研究左归丸含药血清对培养大鼠BMSCs成骨向诱导分化过程中Cx43表达及GJIC功能的影响。方法：密度梯度离心法制取BMSCs培养传代,通过血清药理学方法,设立正常对照A组、左归丸含药血清B组、缝隙连接阻滞C组。培养过程中进行细胞形态学观察;第14天分别采用RT-PCR、FRAP技术检测Cx43、β-catenin、BGPmRNA表达及细胞GJIC功能;第21天茜素红染色检测矿化结节形成情况。结果：左归丸含药血清能明显促进Cx43、β-catenin、BGPmRNA表达,与A组比较,P〈0.05;加用AGA后,Cx43、β-cateninmRNA表达无明显影响,而BGPmRNA明显下降,与B组比较,P〈0.05。FRAP检测镜下观察B组细胞内荧光恢复明显,其平均荧光恢复率与A、C组比较差异显著,P〈0.01。钙结节相对面积定量分析,B组显著高于A、C组,P〈0.05。结论：左归丸含药血清能增强BMSCs成骨向诱导分化过程中Cx43表达及GJIC功能,并与β-catenin、BGP基因在此过程中发挥协同作用。这可能是其促进BMSCs成骨分化成熟的重要机制。Objective： To investigate the effect of Zuogui Pill medicated serum on the expressions of Cx43 and GJIC function during the osteogenic differentiation of cultured rat BMSCs. Methods： BMSCs were isolated from rats with density gradient separation method and then subcultured in vitro. Cells were divided into normal group A,serum group B and gap junction blocker group C by serum pharmacological method. First of all,cell morphology was observed during culture. Secondly,RT-PCR technology was used to detect the mRNA expressions of Cx43,β-catenin and BGP and the function of GJIC was assessed by FRAP technology at day 14. At last,the formation of mineralized nodules were detected by alizarin red staining at day 21. Results： The mRNA expressions of Cx43,β-catenin and BGP in group B were greatly increased,which compared with group A,P〈0. 05. After adding AGA,the expressions of Cx43 and β-catenin mRNA had no significant effect,while BGPmRNA decreased significantly,which compared with group B,P〈0. 05. Confocal microscopy observation showed that the fluorescence of B group was significantly increased,and its mean recovery rate was significantly different with A and C groups,P〈0. 01. Quantitative analysis of relative area of calcium nodules showed that B group was significantly higher than that of A or C group,P〈0. 05. Conclusion： Zuogui Pill medicated serum can enhance Cx43 mRNA expression and GJIC function during the osteogenic differentiation of cultured rats BMSCs and at the same time,β-catenin and BGP genes play a synergistic role in this process. This may be an important mechanism for Zuogui Pill promoting the differentiation and maturation of BMSCs.

关 键 词：左归丸 含药血清 BMSCS CX43 GJIC 

分 类 号：R285[医药卫生—中药学]