补肾强督方含药血清对强直性脊柱炎OPG/RANKL通路的作用

目的探讨补肾强督方对强直性脊椎炎(ankylosing spondylitis,AS)骨保护素和核转录因子-κB受体活化因子配基(osteoprotegerin receptor activator for nuclear factorκB ligand,OPG/RANKL)通路的作用机制。方法选取2009年1-5月卫生部中日友好医院中医风湿病科门诊和住院活动期AS患者30例,予补肾强督方,每天1剂,早晚分两次口服,连续3个月;另选取健康志愿者30名。采集治疗前后患者血清及健康人血清。将成骨细胞hFOB1.19分为患者治疗前组、治疗后组及健康人组(对照组)。各组细胞分别于含相应血清的基础培养基中进行培养。取成骨细胞培养上清液,采用ELISA法检测成骨细胞OPG/RANKL含量及mRNA表达,采用RT-PCR检测OPG/RANKL蛋白表达。结果与对照组比较,治疗前组成骨细胞OPG含量、OPG mRNA及蛋白表达、OPG/RANKL mRNA及蛋白表达均降低,RANKL mRNA表达升高,差异均有统计学意义(P<0.05,P<0.01);与治疗前组比较,治疗后组OPG含量、OPGmRNA及蛋白表达、OPG/RANKL mRNA及蛋白表达升高,RANKL mRNA表达降低,差异均有统计学意义(P<0.05,P<0.01)。结论 AS患者血清能直接抑制成骨细胞OPG表达、促进RANKL表达、下调OPG与RANKL比率,补肾强督方治疗后的血清能直接促进成骨细胞表达OPG,抑制表达RANKL,上调OPG与RANKL比率,补肾强督方可能是通过直接上调成骨细胞OPG/RANKL而抑制破骨细胞分化和功能,达到促进骨生成,抑制骨吸收。

The Function of Bushen Qiangdu Recipe Containing Serum in OPG/RANKL Pathway of Ankylosing Spondylitis Patients

Objective To study the mechanism of Bushen Qiangdu Recipe(BQR) for regulating osteoprotegerin receptor activator for nuclear factor κ B ligand(OPG/RANKL) pathway in ankylosing spondylitis(AS).Methods Thirty active AS inpatients or outpatients were recruited from Department of CM Rheumatology,China-Japan Friendship Hospital from January to May 2009.All patients were treated with BQR for 3 successive months,one dose daily,once in the morning and once in the evening.Besides,30 healthy volunteers were recruited.The serum of patients and volunteers were collected.The osteoblast cell lines hFOB1.19 were divided into 3 groups:the pre-treatment group,the post-treatment group,and the healthy volunteer group(as the control group).All cell lines were cultured by corresponding culture medium containing each serum.The supernatant from osteoblast cell lines was collected.The protein content of OPG/RANKL was detected using ELISA,and the protein expression of OPG/RANKL was detected using RT-PCR.Results Compared with the control group,the OPG content,the mRNA and protein expressions of OPG,and the mRNA and protein expressions of OPG/RANKL all decreased,while the mRNA expression of RANKL increased in the pre-treatment group,showing statistical difference(P<0.05,P<0.01).Compared with the pre-treatment group,the OPG content,the mRNA and protein expressions of OPG significantly increased,and the mRNA and protein expressions of OPG/RANKL increased,while the mRNA expression of RANKL decreased in the post-treatment group,showing statistical difference(P<0.05,P<0.01).Conclusions AS patients’ serum could directly inhibit the expression of OPG in osteoblasts,promote the expression of RANKL,and down-regulate the OPG/RANKL ratio.BQR containing serum might promote the osteogenesis and inhibit the bone resorption possibly through directly up-regulating the OPG/RANKL ratio in osteoblast,thus inhibiting the differentiation and function of osteoclast.