CAJ | 학술논문

目的:观察复方芪麝片(FFQSP)对直立体位大鼠腰椎骨形成的作用,探讨可能的作用机制.方法:90只SD大鼠随机分为三大组,正常组不进行处理,普通饲养笼喂养;模型组和FFQSP组,建立长期直立体位大鼠模型.每大组再随机分为5、7和9个月组,FFQSP组分别在造模后4、6、8个月开始应用FFQSP(将片剂研磨溶于生理盐水)灌胃,连续灌胃1个月;模型组和正常组用同量生理盐水灌胃1个月.所有动物于相应时间点取整个腰椎进行检测,行L5椎体藏红-固绿染色及Ⅰ型胶原(type Ⅰ collegan,ColⅠ)免疫组织化学染色,余下腰椎椎体应用实时荧光定量聚合酶链反应(real time reverse transcription polymerass chain reaction,real time RT-PCR)技术检测Ⅰ型胶原α2(type Ⅰ collegan α2,Col1α2)、转化生长因子beta1(transform growth factor beta1,TGF-β1)和runt相关转录因子2(runt-related transcription factor2,Runx2)基因的表达.结果:藏红染色显示,与正常组比较,三个时间点模型组及FFQSP组椎体骨小梁均增加,FFQSP组在5个月和7个月时较模型组略有增加;在9个月时较模型组略有减少.ColⅠ免疫组织化学染色显示三个时期模型组骨小梁阳性染色数目较正常组均增加,FFQSP组骨小梁阳性染色范围较正常组、模型组均增加.Real time RT-PCR示,与模型组比,5个月FFQSP组Col1α2、TGF-β1基因表达显著下调(P<0.05,P<0.01),与正常组比,仅上调Runx2的表达(P<0.01).7个月FFQSP组Col1α2、TGF-β1、Runx2基因表达较正常组和模型组都显著上调.9个月FFQSP组与模型组比较,Col1α2基因表达显著下调(P<0.05),而TGF-β1和Runx2基因表达均显著上调(P<0.05);与正常组比,三个基因表达均显著上调(P<0.01).结论:复方芪麝片对直立大鼠的椎体成骨有增强作用,以中后期( 7个月、9个月)效应显著,其疗效可能与TGF-β1基因水平的上调有关.
목적:관찰복방기사편(FFQSP)대직입체위대서요추골형성적작용,탐토가능적작용궤제.방법:90지SD대서수궤분위삼대조,정상조불진행처리,보통사양롱위양;모형조화FFQSP조,건립장기직입체위대서모형.매대조재수궤분위5、7화9개월조,FFQSP조분별재조모후4、6、8개월개시응용FFQSP(장편제연마용우생리염수)관위,련속관위1개월;모형조화정상조용동량생리염수관위1개월.소유동물우상응시간점취정개요추진행검측,행L5추체장홍-고록염색급Ⅰ형효원(type Ⅰ collegan,ColⅠ)면역조직화학염색,여하요추추체응용실시형광정량취합매련반응(real time reverse transcription polymerass chain reaction,real time RT-PCR)기술검측Ⅰ형효원α2(type Ⅰ collegan α2,Col1α2)、전화생장인자beta1(transform growth factor beta1,TGF-β1)화runt상관전록인자2(runt-related transcription factor2,Runx2)기인적표체.결과:장홍염색현시,여정상조비교,삼개시간점모형조급FFQSP조추체골소량균증가,FFQSP조재5개월화7개월시교모형조략유증가;재9개월시교모형조략유감소.ColⅠ면역조직화학염색현시삼개시기모형조골소량양성염색수목교정상조균증가,FFQSP조골소량양성염색범위교정상조、모형조균증가.Real time RT-PCR시,여모형조비,5개월FFQSP조Col1α2、TGF-β1기인표체현저하조(P<0.05,P<0.01),여정상조비,부상조Runx2적표체(P<0.01).7개월FFQSP조Col1α2、TGF-β1、Runx2기인표체교정상조화모형조도현저상조.9개월FFQSP조여모형조비교,Col1α2기인표체현저하조(P<0.05),이TGF-β1화Runx2기인표체균현저상조(P<0.05);여정상조비,삼개기인표체균현저상조(P<0.01).결론:복방기사편대직립대서적추체성골유증강작용,이중후기( 7개월、9개월)효응현저,기료효가능여TGF-β1기인수평적상조유관.
Objective:To investigate the effect and potential mechanism of Fu Fang Qi She Pian(FFQSP) on upright rats′ lumbar vertebral bone formation.Method:Ninety SD rats were randomly divided into three groups. The rats in the normal control group were raised in normal cage with no intervention.The rats in untreated and FFQSP groups were induced to upright posture.Every group was randomly divided into three subgroups:5 months,7 months and 9 months group.Rats in FFQSP groups were gastroentericly administrated with FFQSP at 4,6,8 months for one month after surgery while those in model and control group were treated with normal saline.Ten rats were sacrificed at 5,7 and 9 months after surgery and all lumbar vertebre were harvested for detection.Safranine O/fast green staining and ColⅠ immunohistostaining were performed on L5 vertebrae.Col1α2、TGF-β1 and Runx2 mRNA expression in other vertebre were detected by real time reverse transcription polymerass chain reaction(RT-PCR).Result:Safranin O/Fast green staining showed bone trabecular density increased in untreated groups.BV/TV rate raised in 9 months of untreated group compared to normal control group,and it went up in 5,7 months of FFQSP groups compared to untreated groups.Trabecular numbers increased in 5 and 9 months of untreated groups compared to those in the normal group when it increased in 5 and 7 months of FFQSP groups compared to those in untreated groups.Immunohistostaining showed increased numbers of bone trabeculae with Col Ⅰ positive staining after surgery,which extended in FFQSP groups compared to that in control and model groups at each time points.Real time RT-PCR showed Col1α2 and TGF-β1 mRNA expression significantly down-regulated in FFQSP group compared to those in model group at 5 months(P<0.05,P<0.01).However,only Runx2 expression was up-regulated compared with normal group(P<0.01).Col1α2,TGF-β1 and Runx2 gene expression increased in FFQSP groups at 7 months compared with those in control and model groups at the same time point.Col1α2 mRNA expression lowered down in 9 months of FFQSP group compared with that in the model group(P<0.05).While compared with control group,up-regulation expression of all three genes significantly were evidenced(P<0.01).Conclusion:Upright posture may increase rats′ vertebral bone formation which may be strengthened by FFQSP especially at mid-late phase(7- and 9-month groups).The mechanism may be related to the upregulation of TGF-β1 gene level.