摘要
目的分析Ras相关蛋白1(Ras-associated protein 1,Rap1)在非创伤性股骨头坏死(non-traumatic osteonecrosis of the femoral head,NONFH)患者骨组织坏死与正常区域的表达差异。方法分别收集NONFH和创伤性股骨头坏死(traumatic osteonecrosis of the femoral head,TONFH)患者经髋关节置换术后的股骨头组织各30例。NONFH组男19例,女11例;年龄26~69岁,平均49.6岁,国际骨循环协会(ARCO)Ⅲb期8例,Ⅳ期10例,Ⅴ期12例;TONFH组男16例,女14例;年龄37~68岁,平均54.2岁,ARCOⅢb期11例,Ⅳ期9例,Ⅴ期10例。两组间性别及年龄差异无统计学意义(P>0.05),ARCO分期构成比差异亦无统计学意义(Z=-0.769,P=0.442)。取NONFH骨标本坏死区(A区)和正常区(B区),TONFH骨标本正常区域(B’区)(对照组)。采用Western blot、实时荧光定量PCR分析3个区域骨组织中Rap1、血管内皮生长因子(VEGF)、磷脂酰肌醇3-激酶(PI3K)、Akt蛋白及相应mRNA表达差异,HE和免疫组化染色观察各区域的形态学改变。结果Western blot检测结果显示,B区、B’区间Rap1、VEGF、PI3K及Akt蛋白相对表达量差异均无统计学意义(P>0.05)。而A区Rap1、VEGF、PI3K及Ak蛋白相对表达量均较B区减少,差异有统计学意义(P<0.05)。实时荧光定量PCR检测结果显示,与B区比较,A区Rap1、VEGF、PI3K、Akt mRNA相对表达量均较B区降低,差异有统计学意义(P<0.05);B区、B’区两组间Rap1、VEGF、PI3K及Akt mRNA相对表达量差异均无统计学意义(P>0.05)。HE和免疫组化染色:NONFH坏死区域(A区)软骨细胞减少、细胞核消失,软骨下骨小梁断裂,骨小梁增厚,出现空骨陷窝;坏死区周围新生毛细血管和纤维组织构成的肉芽组织增生爬行,但Rap1、VEGF、PI3K和Akt在此区域阳性表达较正常区域弱。同时Rap1、PI3K、Akt在A、B两区域的骨小梁中均有阳性表达,表达强度相似。B区域微动脉、微静脉内膜及周围间质细胞膜上可见Rap1、VEGF、PI3K及Akt均有强阳性表达,但A区较B�
Objective To investigate the difference in the expression of Ras-associated protein 1(Rap1)in necrotic and healthy areas of non-traumatic osteonecrosis of femoral head(NONFH)patients.Methods Femoral head tissue samples from 30 cases of NONFH and 30 cases of traumatic osteonecrosis of the femoral head(TONFH)were collected after hip replacement surgery,respectively.No significant difference of Association Research Circulation Osseous(ARCO)staging was found between the NONFH and the TONFH groups(Z=-0.769,P=0.442).In the NONFH group,8 patients were ARCO stageⅢb,10 were stageⅣ,and 12 were stageⅤ,while in the TONFH ground,11 patients were ARCO stageⅢb,9 were stageⅣ,and 10 were stageⅤ.There were 19 males and 11 females in the NONFH group,with an average age of 49.6 yr.(26-69 yr.),and 16 males and 14 females in the TONFH group,with an average age of 54.2 yr.(37-68 yr.).There was no significant difference in gender or age between the two groups(P>0.05).Specimens were collected from different bone areas,including those from the necrotic areas(area A)and the healthy areas(area B)of the NONFH group,and those from the healthy areas(area B’)of the TONFH group,i.e.,the control group.Western blot and quantitative real-time reverse transcription PCR(qRT-PCR)were used to analyze the different expression of Rap1,vascular endothelial growth factor(VEGF)protein,phosphoinositide 3-kinase(PI3 K),and Akt protein and their corresponding mRNA in the three areas of bone tissue.HE staining and immunohistochemisty staining were done in order to observe the morphological changes of each area.Results Western blot results indicated that there was no statistical difference in the relative expression of Rap1,VEGF,PI3 K,and Akt proteins(P>0.05).The relative expressions of Rap1,VEGF,PI3 K,and Akt proteins in the area A were lower than those in the area B and the difference was statistically significant(P<0.05).qRT-PCR results showed that the relative expressions of Rap1,VEGF,PI3 K and Akt mRNA in area A were lower than those of area B
作者
胡星荣
周明旺
柳海平
李盛华
陈彦同
刘一飞
王鹏志
赵秋玥
肖振
HU Xing-rong;ZHOU Ming-wang;LIU Hai-ping;LI Sheng-hua;CHEN Yan-tong;LIU Yi-fei;WANG Peng-zhi;ZHAO Qiu-yue;XIAO Zhen(Gansu University of Chinese Medicine,Lanzhou 730000,China;Department of Orthopaedics,Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730050,China;Department One of Arthrology,Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730050,China)
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2021年第3期452-457,共6页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.81860861、No.81560782)资助。
关键词
非创伤性股骨头坏死
Ras相关蛋白1
血管内皮生长因子
病理
Non-traumatic osteonecrosis of femoral head
RAS-associated protein 1
Vascular endothelial growth factor
Pathology
