基于调控破骨-成骨细胞分化探究骨痿灵治疗骨质疏松症作用机制  

Study on the mechanisms of Guweiling in the treatment of osteoporosis based on the regulation of osteoclast-osteoblast differentiation

作　　者：靳雯棋 王晶 王宇 杨兰 张予心 付宝玉 JIN Wenqi;WANG Jing;WANG Yu;YANG Lan;ZHANG Yuxin;FU Baoyu(Research Center of Traditional Chinese Medicine,Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130021,China;College of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学附属医院中医药研究中心,长春130021 [2]长春中医药大学中医学院,长春130117

出　　处：《吉林中医药》2025年第1期80-86,共7页Jilin Journal of Chinese Medicine

基　　金：国家自然科学基金资助项目(82104902);吉林省科技发展计划项目(YDZJ202301ZYTS337);吉林省教育厅科学技术研究规划项目(JJKH20230955KJ)。

摘　　要：目的探讨骨痿灵对破骨-成骨细胞和骨丢失小鼠作用机制。方法骨痿灵对RANKL诱导的破骨细胞进行预处理,分别检测破骨细胞TRAP染色、活性,分化特异性标志因子及转录因子的表达变化。通过ALP染色和茜素红S染色检测骨痿灵对成骨细胞早期和晚期分化水平,通过RT-PCR明确成骨细胞分化标志因子及转录因子的表达变化。在体内水平通过HE染色、骨微结构和骨代谢因子显示骨痿灵对LPS诱导的小鼠骨丢失的影响。结果与RANKL阳性对照组比较,骨痿灵组破骨细胞数量显著降低(P<0.05);骨痿灵组内未出现细胞毒性作用(P>0.05);骨痿灵干预后,破骨细胞生成过程中特异性标志物基因TRAP、CtsK、MMP-9、OCSAR和转录因子NFATc1和c-Fos表达降低(P<0.05)。骨痿灵干预对成骨细胞存活率无明显影响(P>0.05);与对照组比较,骨痿灵组ALP染色阳性细胞群增加,深红色矿化结节数量和密度增加(P<0.05);与对照组比较,成骨细胞标志物的基因Runx2、Osx、Col I、OCN表达提高,RANKL表达降低(P<0.05)。与模型组比较,骨痿灵组HE染色发现骨小梁结构恢复和骨基质连接良好;与模型组比较,骨痿灵组骨体积分数和骨小梁数量提高、骨小梁分离度降低(P<0.05);与模型组比较,小鼠血钙和磷上调、β-CTX下降(P<0.05)。结论骨痿灵可通过抑制破骨细胞生成、促进成骨细胞分化,改善骨组织结构,减轻或延缓骨质疏松症。Objective To investigate the effects of Guweiling(GWL)on osteoclast-osteoblasts and bone loss in mice and its possible mechanisms.Methods GWL was used to pretreat RANKL-induced osteoclasts,and their TRAP staining,activity,and expression changes of differentiation-specific marker factors and transcription factors were detected.In addition,the effects of GWL on early differentiation of ALP and late mineralization of calcium nodules in osteoblasts were detected,and its effects on the osteogenic differentiation marker factors and corresponding transcription factors were determined through RT-PCR.Hematoxylin and eosin(H&E),bone microstructure,and bone metabolism factors were used to reveal the in vivo GWL effects on LPS-induced bone loss in mice.Results Compared with the RANKL positive control group,the number of osteoclasts in the GWL group decreased significantly(P<0.05),and there was no cytotoxic effect in the GWL group(P>0.05).After a GWL intervention,the expression of specific marker genes TRAP,CtsK,MMP-9,OCSAR and transcription factors NFATc1 and c-Fos were decreased(P<0.05),while the GWL intervention had no significant effect on the survival rate of osteoblasts(P>0.05).Compared with the control group,the number of ALP stained positive cells and the number and density of dark red mineralized nodules increased in the GWL group(P<0.05).Compared with the control group,the expression of osteoblast marker genes Runx2,Osx,Col I and OCN was increased,while the RANKL expression was decreased(P<0.05).Compared with the model group,HE staining in the GWL group showed that the bone trabecular structure was recovered and the bone matrix connection was good.Compared with the model group,the bone volume fraction and the trabecular number were increased while the trabecular separation was decreased in the GWL group(P<0.05).Compared with the model group,the blood calcium and phosphorus were up-regulated whileβ-CTX was down-regulated(P<0.05).Conclusion Guweiling may improve bone tissue structure,alleviate or delay osteoporosis b

关 键 词：骨质疏松症 骨痿灵 破骨细胞 成骨细胞 分化 

分 类 号：R285[医药卫生—中药学]