二仙汤及其拆方含药血清对成骨细胞分化的影响  

Effects of Erxian Decoction Separate Prescriptions and its Drug-containing Serum on Osteoblasts Osteogenic Differentiation

作　　者：刘波[1,2] 王莹 谢珍[3] 徐彭 Liu Bo;Wang Ying;Xie Zhen;Xu Peng(School of Pharmacy Jiangxi University of Traditional Chinese Medicine, Nanchang 330004;Jiangxi province key laboratory of Chinese Medicine Pharmacology, Nanchang 330004;Zhejiang Institute of Food and Drug Control, Hangzhou 310052)

机构地区：[1]江西中医药大学药学院,南昌330004 [2]江西省中药药理学重点实验室,南昌330004 [3]浙江省食品药品检验研究院,杭州310052

出　　处：《中药药理与临床》2019年第1期22-27,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(81560723;81860802);江西省自然科学基金项目(20114BAB205073;20151BAB205072)

摘　　要：目的:研究二仙汤及其君、臣、佐、使拆方含药血清对成骨细胞分化的影响。方法:实验以成骨细胞系MC3T3-E1细胞为基础,分为空白组、白藜芦醇15mg/kg组、二仙汤整方8g/kg组及依据君臣佐使关系的4个拆方组,即淫羊藿+仙茅3.2g/kg组、淫羊藿+仙茅+巴戟天4.8g/kg组、淫羊藿+仙茅+当归4.8g/kg组、淫羊藿+仙茅+知母+黄柏4.8g/kg组。分别灌胃大鼠,制备含药血清,分别以10%含药血清作用于成骨细胞,以对硝基苯磷酸盐(pNPP)法和碱性磷酸酶(ALP)染色检测成骨细胞ALP活性;以Von Kossa染色法检测成骨细胞矿化结节形成;采用Western blot法检测成骨细胞骨钙素(OC)、骨形成蛋白-2(BMP-2)的表达。结果:与空白组比较,二仙汤整方和各拆方组10%含药血清均能显著增强成骨细胞ALP的活性,其中二仙汤整方组、淫羊藿+仙茅+知母+黄柏组提高成骨细胞ALP活性的作用最为明显,二仙汤整方和各拆方组均可显著提高成骨细胞OC、BMP-2的表达。结论:二仙汤及各拆方含药血清均不同程度地提高成骨细胞ALP活性,增强矿化结节的形成,促进OC、BMP-2的表达,知母、黄柏的作用尤为重要。Objective: To study the effect of serum containing Erxian Decoction and its separate prescriptions on osteoblast osteogenic differentiation. Methods: MC3 T3-E1 cells were divided into 7 groups: Blank group,15 mg/kg Resveratrol group,8 g/kg Erxian Decoction group and four different prescription of Erxian Decoction separate prescriptions groups,namely 3. 2 g/kg Epimedium + Curculigo orchioides Gaertn group,4. 8 g/kg Epimedium + Curculigo orchioides Gaertn + Morinda officinalis group,4. 8 g/kg Epimedium + Curculigo orchioides Gaertn+ Angelica sinensis group and 4. 8 g/kg Epimedium + Curculigo orchioides Gaertn + Anemarrhena asphodeloides Bge + Phellodendron chinensis Schneid group. Rats were orally administered with crude drugs to prepare serum containing drug. 10% serum containing Erxian Decoction and its separate prescriptions were added into the osteoblasts. The activity and content of ALP in osteoblasts were detected by p-Nitrobenzene phosphate( pN PP) and alkaline phosphatase( ALP) staining. The formation of mineralized nodules in osteoblasts was detected by Von Kossa staining. The expressions of osteocalcin( OC) and bone morphogenetic protein-2( BMP-2) in cells were examined by Western Blot. Results: Compared with the blank group,10% serum containing Erxian Decoction and its Separate Prescriptions significantly enhanced the activity of ALP in osteoblasts,and Erxian Decoction and Epimedium + Curculigo orchioides Gaertn + Anemarrhena asphodeloides Bge+ Phellodendron chinensis Schneid group had more significant effect. Erxian Decoction and its separate prescriptions significantly increased the expressions of OC and BMP-2. Conclusion: The serum containing Erxian Decoction and its separate prescriptions can increase ALP activity of osteoblasts,enhance the formation of mineralized nodules,promote the expressions of OC and BMP-2. It shows that the Anemarrhena asphodeloides Bge and Phellodendron chinensis Schneid plays an important role in osteoporosis.

关 键 词：二仙汤 骨质疏松 含药血清 MC3T3-E1 骨形成 

分 类 号：R285[医药卫生—中药学]