补肾通络方调控RANKL-RANK-OPG通路干预破骨细胞相关蛋白的机制研究  

Exploring the mechanism of Bushen Tongluo Formula in regulating osteoclast-related protein through the RANKL-RANK-OPG signaling pathway to intervene bone destruction in collagen-induced arthritis rats

作　　者：李平顺 田杰祥 王丽琴 周孟茹 马鹏程 王涛 魏勇 王钢 LI Ping-shun;TIAN Jie-xiang;WANG Li-qin;ZHOU Meng-ru;MA Peng-cheng;WANG Tao;WEI Yong;WANG Gang(Gansu University of Chinese Medicine,Lanzhou 73000,China;Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 73000,China)

机构地区：[1]甘肃中医药大学附属医院,甘肃兰州730000 [2]甘肃中医药大学,甘肃兰州730000

出　　处：《时珍国医国药》2025年第1期40-49,共10页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金资助项目(81860858,81960832);甘肃省科技厅青年科技基金项目(22JR11RA133);甘肃中医药大学第二附属医院青年项目(ZX-62000004-2022-092);甘肃省名中医传承工作室建设项目【国中医药规财函(2021)242号】。

摘　　要：目的观察补肾通络方对胶原诱导性关节炎(CIA)大鼠的疗效,并探讨可能的机制。方法选用SPF级SD雌雄大鼠,按0.3mL(300μg牛II型胶原乳剂)/只,分别在大鼠双侧后足跖皮内、尾跟部皮下致炎,造模成功后随机分为正常组(生理盐水)、模型组(生理盐水)、甲氨蝶呤组(每周1.04mg/kg)、补肾通络方低剂量组[0.31g/(kg·d)]、中剂量组[0.62g/(kg·d)]、高剂量组[1.24g/(kg·d)],连续灌胃8周,每7天检测1次体重、足跖肿胀度,观察各组大鼠一般状态、骨体积分数(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁间隙(Tb.Sp)、骨小梁数量(Tb.N)、膝关节滑膜病理切片等改变;ELISA法检测血清中RANKL、OPG、TNF-α、IL-6、IL-17表达量;Western blot法测定踝关节RANKL、RANK、OPG表达量;R-T PCR法测定踝关节RANKL、RANK、OPG mRNA表达。结果与正常组比较,模型组大鼠生活状态欠佳,体重下降显著,足跖明显肿胀,关节破坏,滑膜明显增厚,Tb.Sp、RANK、RANKL、TNF-α、IL-6、IL-17明显增加,BV/TV、Tb.Th、Tb.N、OPG明显降低(P<0.05);与模型组比较,甲氨蝶呤组与补肾通络方干预组均可明显改善大鼠生活状态欠佳、体重下降、足跖肿胀、滑膜增厚、关节破坏的情况,增加BV/TV、Tb.Th和Tb.N,减少Tb.Sp,下调RANKL/OPG比例及TNF-α、IL-6、IL-17的表达量,并且补肾通络方高剂量组改善效果更加明显(P<0.05)。结论补肾通络方可有效控制CIA大鼠体内炎性程度和骨破坏的发展,对RA有一定的治疗作用,其机制可能与调节RANKL-RANK-OPG信号通路有关。Objective To observe the therapeutic effect of Bushen Tongluo Formula(BTF)on collagen-induced arthritis(CIA)rats and explore the possible mechanism.Methods Each of the male and female SD rats of SPF grade were injected with 0.3mL(300μg bovine collagen type Ⅱ emulsion)respectively to trigger inflammation in the skin of bilateral hind feet and subcutaneously in the heel of the tail.After successful modeling,the rats were randomly divided into the normal group(normal saline),the model group(normal saline),the methotrexate group(1.04mg/kg·w),the BTF low-dose group(0.31g/kg·d),the BTF medium-dose group(0.62g/kg.d),and the BTF high-dose group(1.24g/kg·d),which were gavaged for 8 weeks.The body weight and foot extension swelling were detected every 7 days.The general conditions,bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular space(Tb.Sp),trabecular number(Tb.N)and pathological section of knee joint were observed.ELISA was used to detect the expressions of RANKL,OPG,TNF-α,IL-6 and IL-17 in serum.Western blot(WB)was used to measure the expressions of RANKL,RANK and OPG in ankle joint.R-T PCR was used to determine the mRNA expressions of RANKL,RANK and OPG in ankle joint.Results Compared with the normal group,the model group showed significant weight loss,marked swelling of foot extension,joint destruction,and synovium thickening.Tb.Sp,RANK,RANKL,TNF-α,IL-6,and IL-17 were significantly increased while BV/TV,Tb.Th,and Tb were significantly decreased(P<0.05).Compared with the model group,the poor living conditions,weight loss,foot extension swelling,synovial thickening and joint destruction of both the methotrexate group and BTF intervention groups were significantly relieved,BV/TV,Tb.Th and Tb.N were increased,while Tb.Sp reduced.The ratio of RANKL/OPG and the expression levels of TNF-a,IL-6 and IL-17 were down-regulated,and the BTF high-dose group showed more significant improvement(P<0.05).Conclusion BTF can effectively inhibit the degree of inflammation and the development of bone destruction i

关 键 词：类风湿关节炎 破骨细胞 骨破坏 RANKL-RANK-OPG信号通路 补肾通络方 分化 

分 类 号：R285.5[医药卫生—中药学]