中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2009年
6期
397-399
,共3页
鲍礼智%韩星海%赵东宝%管剑龙%蔡青%戴生明%施冶青%张兰铃%刘菁
鮑禮智%韓星海%趙東寶%管劍龍%蔡青%戴生明%施冶青%張蘭鈴%劉菁
포례지%한성해%조동보%관검룡%채청%대생명%시야청%장란령%류정
骨保护素%关节炎,实验性%破骨细胞%关节炎,类风湿%基因疗法%细胞因子类%腺相关病毒
骨保護素%關節炎,實驗性%破骨細胞%關節炎,類風濕%基因療法%細胞因子類%腺相關病毒
골보호소%관절염,실험성%파골세포%관절염,류풍습%기인요법%세포인자류%선상관병독
Osteoprotegerin%Arthritis,experimental%Osteoclasts%Gene therapy%Arthritis,rheumatoid%Cvtokines%Adeno-associated virus
目的 探讨携带人护骨素(OPG)基因的重组腺相关病毒(rAAV-hOPG)关节腔内转导对胶原诱导性关节炎(CIA)大鼠关节滑膜OPG、抗酒石酸酸性磷酸酶(TRAP)、血管生长因子(VEGF)基因表达的影响.方法 经牛Ⅱ型胶原诱导建立大鼠关节炎模型,随机分成3组:CIA空白对照组,增强绿色荧光蛋白(EGFP)阴性对照组,OPG治疗组,加上健康对照组,共4组.每组10只,于首次免疫后第25天开始,分别予磷酸盐缓冲液(PBS)、rAAV-EGFP、rAAV-hOPG、PBS,双膝关节腔内注射50μl/侧.第40天取双膝关节滑膜,提取RNA,反转录成cDNA后,使用实时荧光定量聚合酶链反应测定OPG、TRAP、VEGF的相对基因表达量.结果 CIA大鼠较健康鼠,滑膜OPG表达下降32.47%(P<0.05),TRAP升高454.79%(P<0.01),VEGF升高152.74%(P<0.05),rAAV-hOPG感染后,OPG基因转录水平增高128.21%(P<0.01),TRAP表达下降58.79%(P<0.01),VEGF表达下降17.85%(P>0.05).结论 rAAV-hOPG能有效转导关节滑膜组织,对关节破坏有一定的保护作用.
目的 探討攜帶人護骨素(OPG)基因的重組腺相關病毒(rAAV-hOPG)關節腔內轉導對膠原誘導性關節炎(CIA)大鼠關節滑膜OPG、抗酒石痠痠性燐痠酶(TRAP)、血管生長因子(VEGF)基因錶達的影響.方法 經牛Ⅱ型膠原誘導建立大鼠關節炎模型,隨機分成3組:CIA空白對照組,增彊綠色熒光蛋白(EGFP)陰性對照組,OPG治療組,加上健康對照組,共4組.每組10隻,于首次免疫後第25天開始,分彆予燐痠鹽緩遲液(PBS)、rAAV-EGFP、rAAV-hOPG、PBS,雙膝關節腔內註射50μl/側.第40天取雙膝關節滑膜,提取RNA,反轉錄成cDNA後,使用實時熒光定量聚閤酶鏈反應測定OPG、TRAP、VEGF的相對基因錶達量.結果 CIA大鼠較健康鼠,滑膜OPG錶達下降32.47%(P<0.05),TRAP升高454.79%(P<0.01),VEGF升高152.74%(P<0.05),rAAV-hOPG感染後,OPG基因轉錄水平增高128.21%(P<0.01),TRAP錶達下降58.79%(P<0.01),VEGF錶達下降17.85%(P>0.05).結論 rAAV-hOPG能有效轉導關節滑膜組織,對關節破壞有一定的保護作用.
목적 탐토휴대인호골소(OPG)기인적중조선상관병독(rAAV-hOPG)관절강내전도대효원유도성관절염(CIA)대서관절활막OPG、항주석산산성린산매(TRAP)、혈관생장인자(VEGF)기인표체적영향.방법 경우Ⅱ형효원유도건립대서관절염모형,수궤분성3조:CIA공백대조조,증강록색형광단백(EGFP)음성대조조,OPG치료조,가상건강대조조,공4조.매조10지,우수차면역후제25천개시,분별여린산염완충액(PBS)、rAAV-EGFP、rAAV-hOPG、PBS,쌍슬관절강내주사50μl/측.제40천취쌍슬관절활막,제취RNA,반전록성cDNA후,사용실시형광정량취합매련반응측정OPG、TRAP、VEGF적상대기인표체량.결과 CIA대서교건강서,활막OPG표체하강32.47%(P<0.05),TRAP승고454.79%(P<0.01),VEGF승고152.74%(P<0.05),rAAV-hOPG감염후,OPG기인전록수평증고128.21%(P<0.01),TRAP표체하강58.79%(P<0.01),VEGF표체하강17.85%(P>0.05).결론 rAAV-hOPG능유효전도관절활막조직,대관절파배유일정적보호작용.
Objective This study was designed to investigate the expression changes of osteopro-tegerin (OPG), tartrate-resistant acid phosphatase (TRAP) and vascular endothelial growth factor (VEGF) mRNA in collagen induced arthritis(CIA) rats. Methods After CIA was induced in Sprague-Dawley rats, the experimental animals were treated with PBS or rAAV-EGFP or rAAV-hOPG (100 μl/day) intra-articular injection for 10 days. Messenger RNAs (mRNAs) were obtained from CIA synovium 40days after first immun-ization. Reverse transcriptase-polymerase chain reactions (RT-PCR) were carried out to detect the mRNA encoding OPG, TRAP, VEGF and β-actin, which acted as inner control. The genes detected clearly by RT-PCR were quantified using real-time PCR. Results The expression of all genes was confirmed by specific single bands in RT-PCR. Real-time PCR showed that the expression levels of TRAP and VEGF were increased, whereas those of OPG mRNA were decreased in CIA group compared with normal controls. The intra-articular gene transduction markedly increased the gene copies of OPG by 128.21% (P<0.01). The expression change of OPG in synovium also caused the decrease of the expression levels of TRAP and VEGF by 58.79% (P<0.01)and 17.85% (P>0.05) respectively, however, the expression change of VEGF was not statistically significant. Conclusion OPG gene mediated by rAAV can be successfully tranfered to knee joint synovium in vivo. The results of this study suggest that gene transfer using rAAV-OPG may be a feasible and effective therapeutic approach to treat or prevent joint destruction in inflammatory arthritis.
