加味阳和汤含药血清对MC3T3-E1与RAW264.7细胞共育体系的影响  

Effect of Jiawei Yanghe Decoction containing serum on the co-cultivation system of MC3T3-E1 and RAW264.7 cells

作　　者：杨智军 杨文龙 夏汉庭 李威 罗云丰 刘江源 杨凤云[2] YANG Zhijun;YANG Wenlong;XIA Hanting;LI Wei;LUO Yunfeng;LIU Jiangyuan;YANG Fengyun(Jiangxi University of Chinese Medicine,Nanchang 330004,China;Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]江西中医药大学附属医院,江西南昌330006

出　　处：《中国骨质疏松杂志》2022年第9期1342-1346,共5页Chinese Journal of Osteoporosis

基　　金：江西省中医药管理局专项基金资助项目(2020ZGJ01);江西省教育厅科学技术研究项目(GJJ190686);江西中医药大学校级研究生创新专项(JZYC20S15)。

摘　　要：目的 观察加味阳和汤含药血清对MC3T3-E1与RAW264.7细胞共育体系的影响,探究其防治骨质疏松症(OP)肾阳虚证的可能作用机制。方法 制备加味阳和汤含药血清,于Transwell共培养板建立MC3T3-E1与RAW264.7细胞共育体系,设置肾阳虚血清组、空白组、含药血清+肾阳虚血清组及含药血清组。干预48 h后,通过碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRAP)染色观察成骨、破骨分化;运用TRAP和ALP试剂盒分别检测细胞上清液中TRAP、ALP活性;通过Western blot检测各组OPG、RANKL、RANK蛋白表达。结果 与肾阳虚血清组比较,含药血清+肾阳虚血清组ALP活性增加(P<0.05),TRAP活性及OC数目减低(P<0.05,P<0.05),OPG蛋白表达上调(P<0.05),RANKL、RANK蛋白表达下调(P<0.05,P<0.05)。结论 加味阳和汤对OP肾阳虚证有相应治疗作用,其机制可能与增加OPG及降低RANKL、RANK蛋白表达有关。Objective To observe the effect of Jiawei Yanghe Decoction-containing serum on the co-cultivation system of MC3 T3-E1 and RAW264.7 cells, and to explore its possible mechanism of the prevention and treatment of osteoporosis(OP) with kidney-yang deficiency syndrome. Methods Prepare medicated serum of Jiawei Yanghe Decoction, the MC3 T3-E1 and RAW264.7 cell co-cultivation system was established on the Transwell co-culture plate, and the kidney-yang deficiency serum group, blank group, drug-containing serum + kidney-yang deficiency serum group and drug-containing serum group were set up. After the intervention of the co-cultivation system for 48 hours, Alkaline phosphatase(ALP) and tartrate-resistant acid phosphatase(TRAP) staining were used to observe osteogenic and osteoclastic differentiation. TRAP and ALP kits were used to detect TRAP in the cell supernatant. And ALP activity, Western blot detected the expression of OPG, RANKL, RANK protein in each group. Results Compared with the kidney-yang deficiency serum group, the ALP activity of the drug-containing serum + the kidney-yang deficiency serum group increased(P<0.05), the TRAP activity and the number of OC decreased(P<0.05, P<0.05), and the expression of OPG protein was up-regulated(P<0.05, P<0.05), the expression of RANKL and RANK protein was down-regulated(P<0.05, P<0.05). Conclusion Jiawei Yanghe Decoction has corresponding therapeutic effect on OP kidney-yang deficiency syndrome, and its mechanism may be related to the increase of OPG and the decrease of RANKL and RANK protein expressions.

关 键 词：骨质疏松症 加味阳和汤 含药血清 肾阳虚血清 细胞共育 

分 类 号：R285.5[医药卫生—中药学]