中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
33期
5929-5935
,共7页
赵可伟%邱俊林%潘旭枫%梁秀珍%陈小华
趙可偉%邱俊林%潘旭楓%樑秀珍%陳小華
조가위%구준림%반욱풍%량수진%진소화
组织构建%骨组织构建%骨质疏松%中药骨康%核内结合因子a1%核因子κB受体活化因子配体%骨保护蛋白%成骨细胞%细胞培养%中药血清学
組織構建%骨組織構建%骨質疏鬆%中藥骨康%覈內結閤因子a1%覈因子κB受體活化因子配體%骨保護蛋白%成骨細胞%細胞培養%中藥血清學
조직구건%골조직구건%골질소송%중약골강%핵내결합인자a1%핵인자κB수체활화인자배체%골보호단백%성골세포%세포배양%중약혈청학
tissue construction%bone tissue construction%osteoporosis%Chinese medicine Gukang presciption%core binding factor alpha 1%receptor activator of nuclear factor kappa B ligand%osteoprotegerin%osteoblasts%cel culture%serum pharmacology
背景：中药骨康在临床上治疗骨质疏松疗效明确，但其具体作用途径尚不清晰。<br> 目的：假设中药骨康通过调节核内结合因子a1水平，控制其下游基因核因子κB受体活化因子配体和骨保护素表达，起到调控成骨细胞生长发育的作用。<br> 方法：新生24 h内的SD乳鼠用于成骨细胞的分离培养。成年SD雌性大鼠用于制备药物血清，随机分为正常血清组和中药骨康组。2组大鼠按体表面积的方法给予中药骨康方的提取药物和生理盐水灌胃，连续给药1周。最后一次用药后2h行心脏采血，分离血清。原代培养并传至第3代经碱性磷酸酶鉴定取得的大鼠成骨细胞，消化计数铺板并分为2组，以上述血清处理72 h，MTT法检测成骨细胞的增殖率，ELISA方法检测碱性磷酸酶分泌量并以相应吸光度值进行纠正，运用RT-PCR检测2组成骨细胞核内结合因子α1及其下游核因子κB受体活化因子配体和骨保护素mRNA表达情况。<br> 结果与结论：中药骨康组成骨细胞骨保护素和核内结合因子a1 mRNA水平显著高于正常血清组，核因子κB受体活化因子配体蛋白和mRNA水平显著低于正常血清组(P <0.01)。实验结果证实，中药骨康可通过影响核内结合因子a1表达，调控其下游基因核因子κB受体活化因子配体和骨保护蛋白表达和分泌，进而发挥治疗骨质疏松的作用。
揹景：中藥骨康在臨床上治療骨質疏鬆療效明確，但其具體作用途徑尚不清晰。<br> 目的：假設中藥骨康通過調節覈內結閤因子a1水平，控製其下遊基因覈因子κB受體活化因子配體和骨保護素錶達，起到調控成骨細胞生長髮育的作用。<br> 方法：新生24 h內的SD乳鼠用于成骨細胞的分離培養。成年SD雌性大鼠用于製備藥物血清，隨機分為正常血清組和中藥骨康組。2組大鼠按體錶麵積的方法給予中藥骨康方的提取藥物和生理鹽水灌胃，連續給藥1週。最後一次用藥後2h行心髒採血，分離血清。原代培養併傳至第3代經堿性燐痠酶鑒定取得的大鼠成骨細胞，消化計數鋪闆併分為2組，以上述血清處理72 h，MTT法檢測成骨細胞的增殖率，ELISA方法檢測堿性燐痠酶分泌量併以相應吸光度值進行糾正，運用RT-PCR檢測2組成骨細胞覈內結閤因子α1及其下遊覈因子κB受體活化因子配體和骨保護素mRNA錶達情況。<br> 結果與結論：中藥骨康組成骨細胞骨保護素和覈內結閤因子a1 mRNA水平顯著高于正常血清組，覈因子κB受體活化因子配體蛋白和mRNA水平顯著低于正常血清組(P <0.01)。實驗結果證實，中藥骨康可通過影響覈內結閤因子a1錶達，調控其下遊基因覈因子κB受體活化因子配體和骨保護蛋白錶達和分泌，進而髮揮治療骨質疏鬆的作用。
배경：중약골강재림상상치료골질소송료효명학，단기구체작용도경상불청석。<br> 목적：가설중약골강통과조절핵내결합인자a1수평，공제기하유기인핵인자κB수체활화인자배체화골보호소표체，기도조공성골세포생장발육적작용。<br> 방법：신생24 h내적SD유서용우성골세포적분리배양。성년SD자성대서용우제비약물혈청，수궤분위정상혈청조화중약골강조。2조대서안체표면적적방법급여중약골강방적제취약물화생리염수관위，련속급약1주。최후일차용약후2h행심장채혈，분리혈청。원대배양병전지제3대경감성린산매감정취득적대서성골세포，소화계수포판병분위2조，이상술혈청처리72 h，MTT법검측성골세포적증식솔，ELISA방법검측감성린산매분비량병이상응흡광도치진행규정，운용RT-PCR검측2조성골세포핵내결합인자α1급기하유핵인자κB수체활화인자배체화골보호소mRNA표체정황。<br> 결과여결론：중약골강조성골세포골보호소화핵내결합인자a1 mRNA수평현저고우정상혈청조，핵인자κB수체활화인자배체단백화mRNA수평현저저우정상혈청조(P <0.01)。실험결과증실，중약골강가통과영향핵내결합인자a1표체，조공기하유기인핵인자κB수체활화인자배체화골보호단백표체화분비，진이발휘치료골질소송적작용。
BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear. <br> OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts. <br> METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected <br> serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse <br> transcription-PCR in al groups. <br> RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor <br> activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (P<0.01). These findings indicate that Chinese medicine Gukang prescription can modulate the expression of <br> core binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.
