苦参碱对RANKL诱导破骨细胞分化及TRAF6、c-fos、NFATc1表达水平的影响  

Effect of matrine on RANKL-induced osteoclast differentiation and the expression of TRAF6, c-fos and NFATc1

作　　者：李欣 李芳[2] 王媛 张宝玉[1] 张丽洁 LI Xin;LI Fang;WANG Yuan;ZHANG Baoyu;ZHANG Lijie(Endocrine Metabolism and Immune Center,Beijing Luhe Hospital,Capital Medical University,Beijing101149,China;Department of Rheumatology,the Second Hospital of Shanxi Medical University,ShanxiProvince,Taiyuan030001,China)

机构地区：[1]首都医科大学附属北京潞河医院内分泌代谢与免疫性疾病中心,北京101149 [2]山西医科大学第二医院风湿免疫科,山西太原030001

出　　处：《中国医药导报》2020年第1期13-16,32,共5页China Medical Herald

基　　金：北京市通州区科技计划项目（KJ2019CX001-05）;山西省留学人员科技活动择优资助项目（晋财社[2018]123号-8）

摘　　要：目的探讨苦参碱对RANKL诱导破骨细胞分化的影响及其可能的分子学机制。方法 RAW264.7细胞系培养,根据加入破骨细胞诱导剂RANKL及不同剂量苦参碱,分为空白组、诱导组、苦参碱低剂量组(0.5 mg/mL)、苦参碱中剂量组(1 mg/mL)、苦参碱高剂量组(2 mg/mL)。给药后48 h,qPCR、Western blot检测TRAF6、c-fos、NFATc1 m RNA及蛋白的表达,第7天TRAP染色观察多核破骨细胞,第9天qPCR检测Calcr、Ctsk mRNA表达,第12天甲苯胺蓝染色观察骨吸收陷窝。结果不同剂量苦参碱组中多核破骨细胞、骨吸收陷窝、Calcr mRNA、Ctsk m RNA表达量较诱导组减少。苦参碱不同剂量组TRAF6、c-fos、NFATc1 mRNA表达量较诱导组明显下降(P <0.01),且呈剂量依赖性。空白组与苦参碱高剂量组TRAF6、c-fos、NFATc1 mRNA表达量比较差异无统计学意义(P> 0.05)。与诱导组比较,苦参碱低、中、高剂量组TRAF6、c-fos、NFATc1蛋白表达量下降(P <0.05),且呈剂量依赖性。空白组与苦参碱高剂量组TRAF6、c-fos、NFATc1蛋白表达量比较差异无统计学意义(P> 0.05)。结论苦参碱可能通过抑制TRAF6、c-fos、NFATc1的表达,影响RANKL诱导破骨细胞的分化。Objective To investigate the effect of matrine on RANKL-induced osteoclast differentiation and the possible molecular mechanism. Methods RAW264.7 cells were cultured. According to the different addition of drugs(none,osteoclast inducer RANKL and different doses of matrine), they were divided into blank group, induction group, lowdose matrine group(0.5 mg/mL), medium-dose matrine group(1 mg/mL) and high-dose matrine group(2 mg/mL). The expression of TRAF6, c-fos, NFATc1 mRNA and protein were detected by qPCR and Western blot at the 48 thhour after administration. Multinucleated osteoclasts were observed by TRAP on the 7 th day. The expression of Calcr and Ctsk mRNA was detected by qPCR on the 9 thday. The bone resorption lacunae were observed by toluidine blue staining on the 12 thday. Results Compared with induction group, the expression of osteoclasts, bone resorption lacuna, Calcr mRNA and Ctsk mRNA decreased in different dosage matrine group. The expression of TRAF6, c-fos and NFATc1 mRNA in different dosage groups of matrine was significantly lower than that in induction group(P < 0.01), and was dose-dependent. There was no significant difference in the expression of TRAF6, c-fos and NFATc1 mRNA between blank group and high dose group of matrine(P > 0.05). Compared with induction group, the expression of TRAF6, c-fos and NFATc1 protein in matrine low, middle and high dose groups decreased(P < 0.05), and it was dose-dependent. There was no significant difference in the expression of TRAF6, c-fos and NFATc1 between blank group and high dose group of matrine(P > 0.05). Conclusion Matrine may affect RANKL-induced osteoclasts differentiation by inhibiting the expression of TRAF6, c-fos and NFATc1.

关 键 词：苦参碱 RAW264.7细胞 TRAF6 C-FOS NFATc1 

分 类 号：R3[医药卫生—基础医学]