吉非替尼对骨折愈合中Ⅰ、Ⅱ及Ⅹ型胶原基因表达的影响  

Effect of gefitinib on the expression of collagen Ⅰ and Ⅱ and Ⅹ in fracture healing

作　　者：袁功武 兰生辉[2] 曾文波 刘曦明 聂宇[3] YUAN Gong-wu;LAN Shenghui;ZENG Wen-bo;LIU Xi-ming;NIE Yu(Department of Orthopedics, Wuhan Clinical Institute Affiliated to Southern Medical University, Guangzhou 510515, China;Department of Orthopaedics, Wuhan General Hospital of PLA, Wuhan 430070, China)

机构地区：[1]南方医科大学武汉临床医学院,广州510515 [2]中国人民解放军武汉总医院,武汉430070 [3]陆军工程大学军械士官学校门诊部,武汉430075

出　　处：《中国矫形外科杂志》2018年第10期939-944,共6页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81601902);湖北省自然科学基金项目(编号:2015CFB240)

摘　　要：[目的]观察表皮生长因子受体(EGFR)信号通路抑制剂吉非替尼对大鼠股骨骨折愈合中Ⅰ、Ⅱ及Ⅹ型胶原基因表达的影响。[方法]应用大鼠股骨骨折模型,选择48只雄性4周龄SD大鼠随机分为实验组与对照组,实验组通过灌胃予以溶于0.5%甲基纤维素的吉非替尼100 mg/(kg·d),对照组予以0.5%甲基纤维素。术后7、14、21、28 d取血清、骨痂组织及股骨,行ELISA检测血清PINP及CTX,实时荧光定量PCR测定Ⅰ、Ⅱ及Ⅹ型胶原mRNA的表达,同时采用改良Masson染色对骨痂组织中的胶原蛋白进行组织学观察。[结果]Ⅰ型胶原mRNA表达于术后7、14 d高于对照组,Ⅱ型胶原mRNA表达于术后14、21 d高于对照组,Ⅹ型胶原mRNA表达于术后7、14、21 d高于对照组(P<0.05);血清PINP含量于骨7、14、21 d高于对照组,血清CTX含量在第7 d高于对照组(P<0.05);并且改良Masson染色显示实验组第7、14、21 d基质中胶原纤维量多。[结论]通过吉非替尼抑制EGFR信号通路,增加了早期骨折部位骨痂中Ⅰ、Ⅱ及Ⅹ型胶原mRNA的含量,促进了早期骨痂中Ⅰ、Ⅱ及Ⅹ型胶原基因的表达。[Objective] To observe the effect of gefitinib,an inhibitor of epidermal growth factor receptor（EGFR）signaling pathway,on the expression of collagen Ⅰ and Ⅱ and Ⅹ in the femoral fracture healing in rats.[Methods]After a femoral fracture model was created in 48 male 4-week-old SD rats,the animals were randomly divided into the experimental group and control group.The rats in the experimental group received 100mg/kg/day intragastric administration of gefitinib dissolved in 0.5%methylcellulose,while those in the control group were intragastrically given 0.5%methylcellulose only.As soon as the serum,fracture callus and femur specimens were harvested at 7,14,21 and 28 days after operation,the serum levels of PINP and CTX were detected by ELISA,the expression of collagen type Ⅰ and Ⅱ and Ⅹ were measured by real-time quantitative PCR,and the collagen in callus was observed by modified Masson staining.[Results]The experimental group had significantly higher expression of typeⅠcollagen m RNA than the control group at 7 and 14 days after operation（P〈0.05）.In addition,the experimental group got considerably higher expression of type Ⅱ collagen m RNA in at 14 and 21 days than the control group（P〈0.05）.Furthermore,the experimental group proved remarkably higher expression of type Ⅹ collagen m RNA than the control group at 7,14and 21 days（P〈0.05）.Moreover,the experimental group was measured significantly higher levels of serum PINP at 7,14 and 21 days,and higher level of serum CTX at 7 days than the control group（P〈0.05）.In term of histological observation,the experimental group got enhanced collagen fibers in the matrix at 7,14 and 21 days over the control group by the modified Masson staining.[Conclusion]The inhibitory effect of gefitinib on EGFR signaling pathway increase the content of Ⅰ and Ⅱ and Ⅹ collagen type m RNA in the callus from early fracture site,and promoted production of collagen Ⅰ,ⅡandⅩtype in the early callus.

关 键 词：EGFR信号通路 吉非替尼 骨折愈合 Ⅰ型胶原 Ⅱ型胶原 Ⅹ型胶原 

分 类 号：R318[医药卫生—生物医学工程]