[1]章胜权,王煦程,叶福生,等.从脂代谢角度探讨补肾活血方治疗绝经后骨质疏松症的作用机制[J].中医正骨,2022,34(07):1-5.
　ZHANG Shengquan,WANG Xucheng,YE Fusheng,et al.Investigation on the mechanism of Bushen Huoxue Fang(补肾活血方)for treatment of postmenopausal osteoporosis from the view of lipid metabolism[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2022,34(07):1-5.
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从脂代谢角度探讨补肾活血方治疗绝经后骨质疏松症的作用机制()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第34卷
期数:
2022年07期
页码:
1-5
栏目:
基础研究
出版日期:
2022-07-20

文章信息/Info

Title:
Investigation on the mechanism of Bushen Huoxue Fang(补肾活血方)for treatment of postmenopausal osteoporosis from the view of lipid metabolism
作者:
章胜权1; 王煦程2; 叶福生3; 袁文华4
(1.杭州市富阳中医骨伤医院,浙江 杭州 311400; 2.浙江中医药大学,浙江 杭州 310053; 3.浙江萧山医院,浙江 杭州 311200; 4.浙江省中医院,浙江 杭州 310006)
Author(s):
ZHANG Shengquan1; WANG Xucheng2; YE Fusheng3; YUAN Wenhua4
1.Fuyang TCM Orthopedic-Traumatological Hospital,Hangzhou 311400,Zhejiang,China 2.Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China 3.Zhejiang Xiaoshan Hospital,Hangzhou 311200,Zhejiang,China 4.Zhejiang Provincial Hospital of Chinese Medicine,Hangzhou 310006,Zhejiang,China
关键词:
骨质疏松; 绝经后;  补肾活血方;  脂类代谢;  小鼠;  动物实验
Keywords:
osteoporosis; postmenopausal;  Bushen Huoxue Fang;  lipid metabolism;  mice;  animal experimentation
摘要:
目的:从脂代谢角度探讨补肾活血方治疗绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)的作用机制。方法:将15只9周龄SPF级健康雌性C57BL/6J小鼠随机分为3组,每组5只。模型组和补肾活血方组小鼠结扎两侧输卵管后,剪去卵巢进行PMOP造模; 假手术组小鼠结扎两侧输卵管后剪除卵巢旁部分脂肪组织。从造模术后第2天开始,补肾活血方组使用折合生药浓度为1.5 g·mL-1的补肾活血方浓缩剂灌胃,每次0.4 mL,每天1次; 假手术组和模型组小鼠以等量生理盐水灌胃。3组小鼠均每天灌胃1次,连续灌胃8周。药物干预结束后第2天,将所有小鼠脱颈处死,取股骨远端进行观察。以Micro-CT进行骨组织微结构观察,通过阿尔新蓝-苏木素染色观察髓腔中脂肪空泡数量,通过免疫组织化学染色观察骨组织中脂肪酸结合蛋白4(fatty acid-binding protein 4,FABP4)表达情况。结果:①一般情况。实验期间小鼠饮食、精神状况正常,活动规律,至处死前无小鼠死亡。②骨组织微结构观察结果。Micro-CT图像显示,与假手术组相比,模型组小鼠股骨骨密度明显降低,骨小梁减少; 补肾活血方组小鼠的股骨骨密度和骨小梁数量与假手术组相当。假手术组、模型组、补肾活血方组小鼠的股骨骨密度比较,差异有统计学意义[(0.033±0.009)g·cm-3,(0.003±0.001)g·cm-3,(0.022±0.009)g·cm-3,F=14.240,P=0.005]; 模型组的股骨骨密度低于假手术组和补肾活血方组(P=0.046,P=0.050); 补肾活血方组和假手术组的股骨骨密度比较,差异无统计学意义(P=0.203)。③髓腔中脂肪空泡数量观察结果。阿尔新蓝-苏木素染色结果显示,模型组髓腔内脂肪空泡数量较假手术组增加; 补肾活血方组髓腔内脂肪空泡数量少于模型组,与假手术组接近。④骨组织中FABP4表达情况检测结果。假手术组、模型组、补肾活血方组股骨组织中的FABP4含量比较,差异有统计学意义[(1.287±0.015)%,(11.897±0.703)%,(0.990±0.162)%,F=800.200,P=0.000]。模型组的FABP4含量高于假手术组和补肾活血方组(P=0.001; P=0.001),假手术组的FABP4含量高于补肾活血方组(P=0.034)。结论:通过调控FABP4表达,抑制骨髓间充质干细胞向脂肪细胞分化,可能是补肾活血方治疗PMOP的作用机制之一。
Abstract:
Objective:To explore the mechanism of Bushen Huoxue Fang(补肾活血方,BSHXF)in treatment of postmenopausal osteoporosis(PMOP)from the view of lipid metabolism.Methods:Fifteen 9-week-old healthy SPF-grade female C57BL/6J mice were randomly divided into model group,BSHXF group and sham-operated group,5 cases in each group.The mice in model group and BSHXF group were subjected to bilateral tubal ligation,followed by removing ovaries for inducing PMOP; while the ones in sham-operated group underwent bilateral tubal ligation,followed by removal of peri-ovarian adipose tissues.From the 2nd day after the modeling operation,the mice in BSHXF group were intragastric administrated with BSHXF concentrate(equivalent to the crude drug with concentration of 1.5 g/mL),once a day,0.4 ml at a time; while the ones in sham-operated group and model group with the same dose of normal saline.All mice in the 3 groups were intragastric administrated once a day for consecutive 8 weeks.On day 2 after the end of drug intervention,all mice were executed by cervical dislocation and their distal femurs were harvested for observation.The bone tissue microstructure was observed by micro-CT scanning,and the number of fatty vacuoles in medullary cavity was observed via alcian blue-hematoxylin(ABH)staining,moreover,the expression of fatty acid-binding protein 4(FABP4)in bone tissues was detected by using immunohistochemical staining.Results:①During the experiment,all mice were in normal mental states with euphagia and regular activities,and no death occurred before the execution.②As demonstrated by the Micro-CT images,the femur bone mineral density(BMD)was obviously declined and the bone trabecula decreased in model group compared with those in sham-operated group; while the femur BMD and bone trabecula number in BSHXF group were similar to those of sham-operated group.There was statistical difference in femur BMD between the 3 groups(0.033±0.009,0.003±0.001,0.022±0.009 g/cm(3),F=14.240,P=0.005).The femur BMD was lower in model group compared to sham-operated group and BSHXF group(P=0.046,P=0.050),and there was no statistical difference in femur BMD between BSHXF group and sham-operated group(P=0.203).③The results of ABH staining showed that the number of fatty vacuoles in medullary cavity increased in model group compared to sham-operated group,while it was less in BSHXF group compared to model group,and was similar to that of sham-operated group.④There was statistical difference in the level of FABP4 in femur tissues between the 3 groups(1.287±0.015,11.897±0.703,0.990±0.162%,F=800.200,P=0.000).The level of FABP4 was higher in model group compared to sham-operated group and BSHXF group(P=0.001; P=0.001),and was lowest in BSHXF group(P=0.034).Conclusion:BSHXF can inhibit the differentiation of bone marrow mesenchymal stem cells into adipocytes through regulating the expression of FABP4,which may be one of its mechanisms for treatment of PMOP.

参考文献/References:

[1] 王柄棋,孙雨晴,陈翔,等.绝经后骨质疏松症药物治疗的现状与思考[J].中国骨质疏松杂志,2017,23(6):818-823.
[2] 中华中医药学会.绝经后骨质疏松症(骨痿)中医药诊疗指南(2019年版)[J].中医正骨,2020,32(2):1-13.
[3] 许兵,方剑利,刘慧,等.补肾活血方对去势大鼠骨质疏松的影响[J].中华骨质疏松和骨矿盐疾病杂志,2011,4(3):177-182.
[4] 王龙梅,刘康佳,刘子欣,等.脂代谢及相关疾病与骨质疏松症关系研究进展[J].中国骨质疏松杂志,2021,27(5):767-770.
[5] HU X,MA S,YANG C,et al.Relationship between senile osteoporosis and cardiovascular and cerebrovascular diseases[J].Exp Ther Med,2019,17(6):4417-4420.
[6] HU L,YIN C,ZHAO F,et al.Mesenchymal stem cells:cell fate decision to osteoblast or adipocyte and application in osteoporosis treatment[J].Int J Mol Sci,2018,19(2):360.
[7] PAGNOTTI G M,STYNER M,UZER G,et al.Combating osteoporosis and obesity with exercise:leveraging cell mecha-nosensitivity[J].Nat Rev Endocrinol,2019,15(6):339-355.
[8] MURUGANANDAN S,GOVINDARAJAN R,SINAL C J.Bone marrow adipose tissue and skeletal health[J].Curr Osteoporos Rep,2018,16(4):434-442.
[9] FAZELI P K,HOROWITZ M C,MACDOUGALD O A,et al.Marrow fat and bone—new perspectives[J].J Clin Endocrinol Metab,2013,98(3):935-945.
[10] MURUGANANDAN S,SINAL C J.The impact of bone marrow adipocytes on osteoblast and osteoclast differentiation[J].IUBMB Life,2014,66(3):147-155.
[11] MURUGANANDAN S,DRANSE H J,ROURKE J L,et al.Chemerin neutralization blocks hematopoietic stem cell os-teoclastogenesis[J].Stem Cells,2013,31(10):2172-2182.
[12] SHU L,HOO R L,WU X,et al.A-FABP mediates adaptive thermogenesis by promoting intracellular activation of thyroid hormones in brown adipocytes[J].Nat Commun,2017,8:14147.
[13] JUREK S,SANDHU M A,TRAPPE S,et al.Optimizing adipogenic transdifferentiation of bovine mesenchymal stem cells:a prominent role of ascorbic acid in FABP4 induction[J].Adipocyte,2020,9(1):35-50.
[14] FURUHASHI M,TUNCMAN G,GÖRGÜN C Z,et al.Treatment of diabetes and atherosclerosis by inhibiting fatty-acid-binding protein aP2[J].Nature,2007,447(7147):959-965.
[15] ZHAO W,RASHEED A,TIKKANEN E,et al.Identification of new susceptibility loci for type 2 diabetes and shared etiological pathways with coronary heart disease[J].Nat Genet,2017,49(10):1450-1457.
[16] HAO J,ZHANG Y,YAN X,et al.Circulating Adipose fatty acid binding protein is a new link underlying obesity-associated breast/mammary tumor development[J].Cell Metab,2018,28(5):689-705.
[17] LIU Z Z,HONG C G,HU W B,et al.Autophagy receptor OPTN(optineurin)regulates mesenchymal stem cell fate and bone-fat balance during aging by clearing FABP3[J].Autophagy,2021,17(10):2766-2782.
[18] SONG C,TAN P,ZHANG Z,et al.Rev-erb agonism suppresses osteoclastogenesis and prevents ovariectomy-induced bone loss partially via FABP4 upregulation[J].FASEB J,2018,32(6):3215-3228.
[19] 许兵,金红婷,王萧枫,等.补肾活血含药血清对成骨细胞经典Wnt/β-catenin通路的影响研究[J].中国骨伤,2015,28(6):553-558.
[20] XIA C,ZOU Z,FANG L,et al.Bushenhuoxue formula promotes osteogenic differentiation of growth plate chondrocytes through β-catenin-dependent manner during osteoporosis[J].Biomed Pharmacother,2020,127:110170.
[21] XU R,ZENG Q,XIA C,et al.Fractions of Shen-Sui-Tong-Zhi formula enhance osteogenesis via activation of β-catenin signaling in growth plate chondrocytes[J].Front Pharmacol,2021,12:711004.

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备注/Memo

备注/Memo:
基金项目:浙江省中医药科技计划项目(2020ZB208); 杭州市医药卫生科技项目(B20200552); 浙江中医药大学基本科研能力提升项目(2021JKJNTZ017A) 通讯作者:袁文华 E-mail:337979694@qq.com
更新日期/Last Update: 1900-01-01