冬虫夏草提取液抑制破骨细胞分化与缓解去卵巢小鼠骨量流失的研究  

Cordyceps sinensis extract protects against the ovariectomy-induced bone loss via the action on osteoclasts

作　　者：彭伟彪 王婷 冯旭[1] 汪家春 张阵阵 储智勇 PENG Weibiao;WANG Ting;FENG Xu;WANG Jiachun;ZHANG Zhenzhen;CHU Zhiyong(Naval Medical Center of the PLA,Shanghai 200433,China;Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]海军特色医学中心,上海200433 [2]江西中医药大学药学院,江西南昌330004

出　　处：《药学实践与服务》2023年第2期97-105,共9页Journal of Pharmaceutical Practice and Service

基　　金：上海市科学技术委员会技术标准专项项目(16DZ0501400);海军医学研究所基金(16A008)。

摘　　要：目的探讨冬虫夏草提取液(Cordyceps sinensis extract,CSE)对去卵巢小鼠骨量流失的保护作用以及对核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)诱导的破骨细胞分化及其功能的影响。方法从C57BL/6小鼠骨髓中提取巨噬细胞(bone marrow-derived macrophages,BMMs);在破骨细胞分化过程中,加入CSE干预处理,通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色分析破骨细胞数量;将接近成熟的破骨细胞种于羟基磷灰石板,观测骨陷窝面积;使用DAPI和鬼笔环肽对破骨细胞肌动蛋白环(F-actin)进行染色,观测环内细胞核数量和环形态;使用q-PCR检测DC-STAMP、ATP6V0d2、TRAP、CTSK、NFATc1的表达;使用Western blot检测MAPK通路蛋白的表达;构建去卵巢小鼠,每天灌胃给予CSE,治疗6周取小鼠股骨进行形态学分析以及ELISA检测外周血中骨碱性磷酸酶(bone alkaline phosphatase,ALP)、骨钙素(BGP)、TRAP含量。结果CSE显著性地抑制了破骨细胞的分化,且呈剂量依赖性;并且主要作用于破骨细胞分化的早期阶段;也抑制了F-actin环的形成,减少了骨陷窝面积;同时抑制DC-STAMP、ATP6V0d2、TRAP、CTSK、NFATc1的表达和MAPK通路JNK、ERK和P38的激活;CSE治疗也可减缓小鼠的骨量流失,提高血清中ALP、BGP含量,降低TRAP含量。结论CSE通过抑制MAPK通路激活,从而抑制破骨细胞分化及其功能,并对去卵巢小鼠的骨量流失具有良好的保护作用。Objective To explore the effects of Cordyceps sinensis extract(CSE)on osteoporosis and RANKL-mediated osteoclastogenesis.Methods Bone marrow-derived macrophages(BMMs)was isolated from the bone marrow of C57BL/6 mice.CSE was added in osteoclast differentiation.Osteoclasts were stained by tartrate-resistant acid phosphatase(TRAP).The nearly mature osteoclasts were planted on hydroxyapatite plates and the area of bone lacunae was observed by microscope.The F-actin belt was stained by DAPI and phylloeptide and the number of nuclei was observed by confocal microscopy.The expressions of DC-STAMP,ATP6V0D2,TRAP,CTSK,and NFATC1 were detected by q-PCR.The protein expression of the MAPK pathway was detected by Western Blot.The in vivo experiments were carried out by administering CSE to the ovariectomized mice daily through gavage.After 6 weeks of intervention,mouse femurs were taken for morphological analysis.Peripheral blood was taken for ELISA.Results CSE represses osteoclastogenesis,bone resorption,F-actin belts formation,osteoclast specific gene expressions and MAPK signaling pathways in vitro.In vivo study indicated that CSE prevents OVX-induced osteoporosis and preserves bone volume by repressing osteoclast activity and function.It also increases the serum ALP,BGP content,and reduces TRAP content.Conclusion CSE can attenuate osteoclast formation and OVX-induced osteoporosis,suggesting potential clinical therapeutic effects for osteoporosis.

关 键 词：冬虫夏草提取液 破骨细胞 分化 功能 机制 去卵巢小鼠 

分 类 号：R285.5[医药卫生—中药学]