基于RANKL/RANK-NF-κB-NLRP3炎性小体通路探讨强骨饮治疗骨质疏松症模型大鼠的作用机制  

Mechanism of Qiangguyin in Inhibiting Osteoporosis Model Rats Based on the RANKL/RANK-NF-κB-NLRP3 Inflammasome Pathway

作　　者：邓祖跃 李志豪 徐俊峰[3] 黄俊俊 董艳蓉 蒋霞 任艳云[5] DENG Zuyue;LI Zhihao;XU Junfeng;HUANG Junjun;DONG Yanrong;JIANG Xia;REN Yanyun(Zhejiang Institute for Food and Drug Control·Key Laboratory of Drug Contacting Materials Quality Control of Zhejiang Province·NMPA Key Laboratory for Quality Evaluation of Traditional Chinese Medicine,Hangzhou,Zhejiang,China 310052;Zhejiang University of Technology,Hangzhou,Zhejiang,China 310014;Tongde Hospital of Zhejiang Province,Hangzhou,Zhejiang,China 310012;The Second Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine,Hangzhou,Zhejiang,China 310005;No.903 Hospital of PLA,Hangzhou,Zhejiang,China 310022)

机构地区：[1]浙江省食品药品检验研究院·浙江省药品接触材料质量控制研究重点实验室·国家药品监督管理局中成药质量评价重点实验室,浙江杭州310052 [2]浙江工业大学,浙江杭州310014 [3]浙江省立同德医院,浙江杭州310012 [4]浙江中医药大学附属第二医院,浙江杭州310005 [5]中国人民解放军联勤保障部队第903医院,浙江杭州310022

出　　处：《中国药业》2025年第3期42-49,共8页China Pharmaceuticals

基　　金：浙江省中医药科技计划项目[2021ZB178]。

摘　　要：目的探讨强骨饮调节核因子-κB受体激活因子配体/核因子-κB受体激活因子-核因子-κB受体-NOD样受体热蛋白结构域相关蛋白3(RANKL/RANK-NF-κB-NLRP3)炎性小体通路治疗骨质疏松症(OP)的作用机制。方法将100只SD大鼠随机分为正常对照组(A组,等量生理盐水,灌胃给药),OP组(B组,等量生理盐水,灌胃给药),强骨饮低、中、高剂量组[C_(1)组、C_(2)组、C_(3)组,0.83,1.66,3.32 g/(kg·d),灌胃给药],己烯雌酚(DES)组[D组,0.1 mg/(kg·d),灌胃给药],吡咯烷二硫代氨基甲酸铵(PDTC)组[E组,0.1 g/(kg·d),腹腔注射],C_(2)组+E组(F组),氟甲基酮(Z-YVAD)组[G组,0.15 mg/(kg·d),尾静脉给药],C_(2)组+G组(H组),各10只。除A组外,其余各组大鼠均行腹部切口双侧去卵巢术,复制OP大鼠模型。连续6周给药后处死大鼠,采用发色底物法测定血清丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力;采用酶联免疫吸附试验(ELISA)法检测血清白细胞介素(IL)-1β、IL-18、IL-6、肿瘤坏死因子-α(TNF-α)、骨形成发生蛋白-2(BMP-2)、Ⅰ型胶原C端肽(CTX-Ⅰ)水平;采用免疫印迹(Western blot)法检测骨组织中RANKL、RANK、NF-κB亚基p65(NF-κB p65)、NLRP3、胱天蛋白酶-1 p20(caspase-1 p20)、消皮素D-N端抗体(GSDMD-N)的蛋白表达水平;采用双能X线扫描仪测定大鼠股骨中段骨密度(BMD);采用Micro-CT仪观察大鼠股骨头的结构,记录骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.sp)、骨体积分数(BV/TV);采用苏木精-伊红(HE)染色法观察各组大鼠骨组织的病理变化。结果与B组比较,C_(1)组、C_(2)组、C_(3)组、D组、E组、F组、G组、H组大鼠的BMD,Tb.N,Tb.Th,BV/TV均显著升高(P<0.05),Tb.sp均显著降低(P<0.05),骨皮质更厚,骨小梁更粗、排列更有序,成骨细胞数量更多,血清CTX-Ⅰ,IL-1β,IL-18,IL-6,TNF-α水平均显著降低(P<0.05),其中F组较E组、H组较G组以上指标均显著更优(P<0.05);C_(1)组、C_(2)组、C_(3)组、D组、F组�Objective To investigate the mechanism of Qiangguyin in regulating RANKL/RANK-NF-κB-NLRP3 inflammasome pathway in the treatment of osteoporosis(OP).Methods A total of 100 SD rats were randomly divided into the normal control group(group A,equal volume of physiological saline,administered by gavage),the OP group(group B,equal volume of physiological saline,administered by gavage),the low-,medium-,and high-dose groups of Qiangguyin[groups C_(1),C_(2),C_(3),0.83,1.66,3.32 g/(kg·d),administered by gavage],the diethylstilbestrol(DES)group[group D,0.1 mg/(kg·d),administered by gavage],the pyrrolidine dithiocarbamate(PDTC)group[group E,0.1 g/(kg·d),intraperitoneal injection],the group C_(2)+group E(group F),the Z-YVAD group[group G,0.15 mg/(kg·d),administered via tail vein],and the group C_(2)+group G(group H),10 mice in each group.Except for rats in group A,the rats in the other groups underwent bilateral ovariectomy through the abdominal incision to replicate OP rat models.Rats were euthanized after six consecutive weeks of administration.The levels of serum malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by the chromogenic substrate method.The levels of serum interleukin(IL)-1β,IL-18,IL-6,tumor necrosis factor-α(TNF-α),bone morphogenetic protein-2(BMP-2),and C-terminal peptide of type Ⅰ collagen(CTX-Ⅰ)were detected by the enzyme-linked immunosorbent assay(ELISA).The expression levels of RANKL,RANK,NF-κB p65,NLRP3,caspase-1 p20,and GSDMD-N antibody protein were detected by the Western blot method.The bone mineral density(BMD)of the mid-femur was measured by dual energy X-ray absorptiometry.The structure of the femur was observed by the Micro-CT scanner.The trabecular number(Tb.N),trabecular number(Tb.N),trabecular separation(Tb.sp),and bone volume/tissue volume(BV/TV)were recorded.The pathological changes of bone tissue in each group were observed by the hematoxylin-eosin(HE)staining method.Results Compared with those in group B,the BMD,Tb.N,Tb.Th,BV/TV significantly increased(P<0.05),while

关 键 词：强骨饮 骨质疏松症 核因子-κB受体激活因子配体 核因子-κB受体激活因子-核因子-κB受体-NOD样受体热蛋白结构域相关蛋白3 

分 类 号：R932[医药卫生—生药学] R285.5[医药卫生—药学]