摘要
目的 :观察中药复方补肾活血液对体外培养新生SD大鼠成骨细胞增殖、分化、矿化的影响。方法 :采用二次酶消化法从新生SD大鼠颅骨获取成骨细胞进行培养 ,取第 3代成骨细胞为实验模型 ,将其分为 4组 ,在各组中分别加入双蒸馏水 (对照组 ) ,低 (2 0mg/L)、中 (4 0mg/L)、高 (80mg/L)浓度的补肾活血中药提取液。用MTT法(波长 5 70nm处A值表示 )测定细胞增殖功能 ;用对硝基苯基磷酸二钠 (PNPP)法测定碱性磷酸酶 (ALP)活性 ;用放免分析方法测定细胞培养液中的骨钙素 (BGP)含量 ;用茜素红染色法 ,在 4 0倍光镜下作矿化结节计数。结果 :中药中、高浓度组的A值在 2 4h、4 8h、72h时明显高于对照组 (P <0 0 1) ,且与药物浓度呈剂量依赖性 ;低浓度组的A值在72h时点明显高于对照组 (P <0 0 5 ) ,在 2 4h、4 8h时点与对照组相比无显著差异 (P >0 0 5 )。不同浓度给药组培养4 8h、72h、96h后 ,ALP(除低浓度组培养 4 8h外 )、BGP均呈不同程度增加 (P <0 0 1)。在中、高浓度组矿化结节数量 /视野 ,显著多于对照组 (P <0 0 1) ,低浓度组与对照组比较无显著差异 (P >0 0 5 )。结论 :补肾活血液具有促进体外培养的新生SD大鼠成骨细胞增殖、分化及矿化作用 ,这种作用呈剂量相关性。
AIM: To investigate the effect for Chinese herbal recipe exctract for replenishing kidney and activating blood circulation (CRERKABC) on cell proliferation, differentiation and mineralization in cultured rat osteoblasts. METHODS: Osteoblasts from craniums of newly born SD rats were cultured in vitro , and the third-passage osteoblasts were exposed to CRERKABC(20 mg/L, 40 mg/L, 80 mg/L) and vehicle, respectively. MMT was used to assess ability of cell proliferation. Activity of akaline phosphatase was assayed by p-nitrophenyl picolinate, and content of osteocalcin in suppernatants was examined by radioimmunoassay. In addition, alizarin bordeaux staining method was used to count mineralzation nodes under optical microscope. RESULTS: ①CRERKABC increased the proliferation of osteoblasts in a dose and time-dependant manner. ②Akaline phosphatase(ALP) activity and osteocalcin content were also increased in CRERKABC-treated groups at 48 h , 72 h, 96 h, except ALP in low concentration CRERKABC group at 48 h( P<0.01 ). ③Amount of mineralized nodes in middle and high concentration CRERKABC-treated groups were obviously higher than that in control group ( P<0.01 ); There was no difference in amount of mineralized nodes between low concentration CRERKABC-treated group and control ( P>0.05 ). CONCLUSION: CRERKABC can promote proliferation, differentiation, maturation and mineralization of the osteoblasts cultured in vitro.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2003年第6期769-772,共4页
Chinese Journal of Pathophysiology
基金
广东省自然科学基金资助项目 (No .990 4 75 )
广州市科技攻关重点项目 (No .2 0 0 2Z3-E5 0 71)资助
关键词
中草药
大鼠
细胞培养
成骨细胞
Drugs, Chinese herbal
Rats
Cell culture
Osteoblasts
