CN113599396B - 天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用 - Google Patents
天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用 Download PDFInfo
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Abstract
本发明公开了天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用,天然化合物为双去甲氧基姜黄素;应用中将人间充质干细胞调配成人间充质干细胞制剂,将双去甲氧基姜黄素制成双去甲氧基姜黄素制剂,构建骨质疏松症哺乳动物模型;连续12周,每天用双去甲氧基姜黄素制剂对哺乳动物模型经口灌胃给药1次,同时每两周1次对哺乳动物模型尾静脉输注人间充质干细胞制剂。本发明通过双去甲氧基姜黄素与人间充质干细胞联用,来改善骨质疏松症机体内环境,进而营造一种有利于人间充质干细胞向成骨细胞分化的微环境,使得所移植的人间充质干细胞趋高效地向成骨细胞定向分化,并抑制破骨细胞形成,限制人间充质干细胞向脂肪细胞分化,从而提高治疗效果。
Description
技术领域
本发明属于医药技术领域,特别涉及天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用,具体涉及双去甲氧基姜黄素联合人间充质干细胞在制备治疗骨质疏松症药物中的应用。
背景技术
骨质疏松症是一种与增龄或雌激素分泌减少相关的退行性骨代谢疾病,表现为骨量减少、骨微结构破坏,导致骨强度降低,骨折风险增加为特征的全身性慢性骨骼疾病。它是世界范围内最常见的骨代谢性疾病,并且,随着全球人口老龄化程度加剧,患病率呈高发态势。 积极防治骨质疏松症已成为世界医学领域亟待解决的重大课题。
骨质疏松症的病因病机极其复杂,是一类涉及多因素的常见难治性慢性重大疾病。人体正常情况下,骨通过持续的破骨细胞的骨吸收和成骨细胞的骨形成来维持矿化平衡及自身结构完整,实现骨重塑过程的动态偶联平衡,而骨重建动态失衡将导致骨质疏松的发生。治疗OP的理想策略是抑制破骨细胞的骨吸收或增加成骨细胞的骨形成(Black DM,Rosen CJ. Postmenopausal osteoporosis. New England Journal of Medicine 2016,374 (3), 254-262; Cauley JA. Osteoporosis: Fracture epidemiology update 2016.Current Opinion in Rheumatology, 2017, 29(2): 150-156.)。但是,目前两类抗骨质疏松症临床一线药物主要有两大类化学药物,即促进骨形成剂(特立帕肽、罗莫珠单抗)和抑制骨吸收剂(双膦酸盐类药物、雌激素)(Goode SC, Beshears JL, Goode RD, et al.Putting the brakes on breaks: osteoporosis screening and fracture prevention.Geriatric Orthopaedic Surgery & Rehabilitation, 2017, 8 (4): 238-243.)。实际上,这些药物均不能促进骨形成,增加骨量和骨密度,达到根治OP目的,仅起到有限的缓解作用,而且,存在诸多安全性问题,轻者导致血压升高、头晕恶心及胃肠不适等不良反应,重者出现闭经、阴道不规则出血、高钙血症、骨坏死,乃至子宫内膜癌、乳腺癌等(Ding Y,Jiang H, Meng B, et al. Sweroside-mediated mTORC1 hyperactivation in bonemarrow mesenchymal stem cells promotes osteogenic differentiation. Journal ofCellular Biochemistry, 2019, 120 (9): 16025-16036; Li H, Xiao Z, Quarles LD,et al. Osteoporosis: mechanism, molecular target, and current status on drugdevelopment. Current Medicinal Chemistry, 2021, 28: 1489-507)。因此,寻找治疗骨质疏松症新药和新型治疗技术成为当前生物医药领域关注的热点。
近些年来,干细胞技术得到空前发展。间充质干细胞(mesenchymal stem cells,MSCs)是最有可能广泛应用于临床一类多能干细胞,其在移植治疗心血管疾病、骨损伤、神经系统及恶性肿瘤等疾病的研究取得了明显进展(Williams AR, Trachtenberg B,Velazquez DL, et al. Intramyocardial stem cell injection in patients withischemic cardiomyopathy: Functional recovery and reverse remodeling.Circulation Research, 2011, 108(7): 792-796; Shi J, Zhang X, Zhu J, et al.Nanoparticle delivery of the bone morphogenetic protein 4 gene to adipose-derived stem cells promotes articular cartilage Arthroscopy:The Journal ofArthroscopic repair in vitro and in vivo. & Related Surgery, 2013, 29(12):2001-2011.),有望成为治疗难治性疾病的新型技术。因此,MSCs移植技术也为骨质疏松的治疗提供了新思路。研究表明,MSCs对骨质疏松有一定的治疗效果(Kiernan J, Hu S,Grynpas MD, et al. Systemic mesenchymal stromal cell transplantation preventsfunctional bone loss in a mouse model of age-related osteoporosis. Stem CellsTranslational Medicine, 2016, 5:683-93; Sui B, Hu C, Zhang X, et al.Allogeneic mesenchymal stem cell therapy promotes osteoblastogenesis andprevents glucocorticoid-induced osteoporosis. Stem Cells TranslationalMedicine, 2016, 5:1238-46.)。但是,MSCs作为成骨细胞和脂肪细胞的祖细胞,机体内的微环境将决定MSCs的命运,即成骨分化还是成脂分化(Chen Q, Shou P, Zheng C, et al.Fate decision of mesenchymal stem cells: adipocytes or osteoblasts Cell Death& Differentiation, 2016, 23 (7):1128-39)。而且,诸多研究发现MSCs移植到骨质疏松症患者体内,其往脂肪细胞分化而不是成骨细胞,从而加重骨质疏松症患者的病情(Hu L,Yin C, Zhao F, et al. Mesenchymal stem cells: cell fate decision toosteoblast or adipocyte and application in osteoporosis treatment.International Journal of Molecular Sciences, 2018,19:360; Jiang Y, Zhang P,Zhang X, et al. Advances in mesenchymal stem cell transplantation for thetreatment of osteoporosis.Cell Proliferation, 2021, 54:e12956)。因此,MSCs用于治疗骨质疏松症仍然存在诸多不确定性,如何规避这些不利因素,找出内在原因,提出新型MSCs治疗策略有重要的临床意义。
发明内容
针对上述问题,本发明的目的在于,提供天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用。
本发明的目的是通过以下技术方案实现的:
天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用。所述天然化合物为双去甲氧基姜黄素,分子式为C19H16O4, 化学结构式如通式(I)所示。
通式(I)
在所述的应用中,将人间充质干细胞用PBS缓冲液调配成人间充质干细胞制剂,将双去甲氧基姜黄素溶解到0.5%羧甲基纤维素钠溶液中制成双去甲氧基姜黄素制剂(0.5%羧甲基纤维素钠溶液的量能保证双去甲氧基姜黄素完全溶解在其中即可),构建骨质疏松症哺乳动物模型;连续12周,每天用双去甲氧基姜黄素制剂对哺乳动物模型经口灌胃给药1次,同时每两周1次对哺乳动物模型尾静脉输注人间充质干细胞制剂200ul(即0.2ml)。本发明通过每天对骨质疏松症哺乳动物模型经口灌胃双去甲氧基姜黄素制剂来调节骨质疏松症动物模型体内微环境,在骨质疏松症体内营造利于人间充质干细胞向成骨细胞分化并限制向脂肪细胞分化的骨形成环境。连续治疗12周后,可极大改善单独使用人间充质干细胞治疗效果,显著增加骨质疏松症哺乳动物模型血清中骨形成相关标志物水平,降低骨吸收标志物水平,明显增加骨密度,改善骨微结构,阻止骨质流失。
进一步的,所述人间充质干细胞制剂中人间充质干细胞的浓度为2×106个/200μl;所述双去甲氧基姜黄素制剂中双去甲氧基姜黄素的使用剂量为100mg/kg/天。所述人间充质干细胞制剂选用对数生长期为2-5代的人间充质干细胞制成。
进一步的,所述骨质疏松症哺乳动物模型为骨质疏松症小鼠动物模型。
进一步的,所述天然化合物可与医学上可接受的药用辅料制成片剂、胶囊剂、丸剂、脂肪乳剂或软膏剂的药物。在制成的药物中所述天然化合物的使用剂量50-200mg/kg/天。
申请人认为改善骨质疏松症机体内在微环境,特别是营造一种有利于向成骨细胞分化的微环境,使得所移植的MSCs趋高效地向成骨细胞定向分化,是确保其对骨质疏松症疗效的有效途径。成骨细胞分化诱导剂可能是改善骨质疏松症机体微环境最有潜力的候选药物。但是,现有的成骨细胞定向诱导剂多为化学合成药物、激素、细胞因子等,这些诱导剂存在诱导效率低、成本高以及毒副作用等问题。譬如,常用的成骨细胞诱导剂地塞米松在体外能诱导干细胞向成骨细胞分化,但在体内可诱发骨质疏松甚至病理性骨折(Zhao M, LiP, Xu H, et al. Dexamethasone-activated MSCs release MVs for stimulatingosteogenic response. Stem Cells International,2018, 2018: 1-12.),从而限制了其临床应用。因此,发掘高效、安全的新型成骨诱导剂对间充质干细胞移植治疗骨质疏松症等骨病有重要的意义。天然化合物具有化学结构新颖、生物活性多样、作用机制独特及成药性好等特点,是新药发现的重要源泉。发明人通过大量筛选工作,从数百种来自滋补强壮、固本扶正作用传统中药来源的天然化合物中,在国际上首次发现双去甲氧基姜黄素(Bisdemethoxycurcumin,BDMC)具有明显的定向诱导人间充质干细胞(human mesenchymalstem cells, hMSCs)向成骨细胞分化的潜力。BDMC是一种多酚类化合物姜黄素的衍生物,以前研究报道BDMC具有抗氧化、抗肿瘤、抗炎、降脂、神经保护、免疫调节等活性(RamezaniM, Hatamipour M, Sahebkar A. Promising anti-tumor properties ofbisdemethoxycurcumin: a naturally occurring curcumin analogue. Journal ofCellular Physiology, 2018, 233(2): 880-887.Xu Y, Hu R, He D, Zhou G, Wu H, XuC, He B, Wu L, Wang Y, Chang Y, Ma R, Xie M, Xiao Z. Bisdemethoxycurcumininhibits oxidative stress and antagonizes Alzheimer's disease by up-regulating SIRT1. Brain and Behavior. 2020;10(7):e01655.Jin F, Chen X, Yan H,Xu Z, Yang B, Luo P, He Q. isdemethoxycurcumin attenuates cisplatin-inducedrenal injury through anti-apoptosis, anti-oxidant and anti-inflammatory.European Journal of Pharmacology. 2020; 874:173026)。而且,发明人在国际上首次建立了BDMC和hMSCs联合治疗骨质疏松症的策略,取得显著疗效,具有巨大临床应用潜力。
本发明的有益效果在于:
本发明通过天然化合物BDMC与 hMSCs联用,来改善骨质疏松症机体内环境,进而营造一种有利于人间充质干细胞(hMSCs)向成骨细胞分化的微环境,使得所移植的人间充质干细胞(hMSCs)趋高效地向成骨细胞定向分化,并抑制破骨细胞形成,限制人间充质干细胞(hMSCs)向脂肪细胞分化,从而提高治疗效果,为骨质疏松治疗提供一种新的、安全、有效的策略。
附图说明
下面结合附图对本发明做进一步详细说明。
图1为BDMC的LC-MS分析与鉴定图:(A)为总离子流图;(B)为质谱图。
图2为不同浓度BDMC处理hMSCs 6天后,成骨细胞早期标志物碱性磷酸酶的表达情况。
图3为不同浓度BDMC对hMSCs的影响:(A)成骨细胞早期标志物碱性磷酸酶的酶比活力(6天);(B)细胞毒性分析(24-72小时)。
图4为BDMC诱导hMSCs向成骨细胞定向分化,成骨细胞早期(6天)及晚期(21天)标志基因的表达情况。
图5为BDMC诱导hMSCs向成骨细胞定向分化,成骨细胞早期(6天)及晚期(21天)标志蛋白的表达情况。
图6为BDMC诱导hMSCs向成骨细胞定向分化21天后成骨特异性标志物钙化结节形成情况。
图7为BDMC和hMSCs治疗骨质疏松症动物模型12周后,股骨骨量及骨微结构的变化情况;(A)股骨的Micro-CT影像图;(B)骨体积分数变化情况;(C)骨矿物质密度变化情况;(D)骨小梁数量变化情况;(E)骨小梁分离度变化情况;(F)骨小梁厚度变化情况。
图8为BDMC和hMSCs治疗骨质疏松症动物模型12周后,血清中骨吸收与骨形成标志物改变情况;(A)骨吸收标志物CTX-1变化情况;(B)骨吸收标志物TRAP的变化情况;(C)骨形成标志物OCN的变化情况;(D)骨形成标志物PINP的变化情况。
图9为BDMC和hMSCs治疗骨质疏松症动物模型12周后,肝组织病理及肝功能的变化情况;(A)肝组织的病理变化;(B)肝功能的变化(AST:谷草转氨酶;ALT:谷丙转氨酶)。
图10为BDMC和hMSCs治疗骨质疏松症动物模型12周后,肾脏组织病理及肝功能的变化情况;(A)肾组织的病理变化;(B)肾功能的变化(Cr:肌酐;BUN:尿素氮)。
图中所示:NG-对照组;MG-模型组;SG-溶剂对照组;ivBG:BDMC-药物治疗组;hMSCs-人间充质干细胞移植治疗组;ivCG:BDMC药物与人间充质干细胞联合治疗组。
具体实施方式
下面由特定的具体实施例说明本发明的实施方式,熟悉此技术的人士可由本说明书所揭露的内容轻易地了解本发明的其他优点及功效。所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1:BDMC的纯度分析与质谱鉴定
经过分析与鉴定,BDMC为黄色结晶体,溶于甲醇、乙醇和二甲基亚砜等有机溶剂。液质联用分析纯度达到98%以上(如图1A所示),质谱分析结果如图1B所示,ESI-MS:m/z309.11[M+H]+。化学分子式为C19H16O4,分子量308,其结构式如图1B所示。
实施例2:BDMC诱导hMSCs向成骨细胞定向分化的作用
1.实验方法:
将BDMC溶解到二甲基亚砜内制成6份浓度分别为5μM、10μM、15μM、20μM、25μM、30μM的BDMC制剂;将hMSCs分成7组并分别用成骨培养液基进行培养,其中一组作为对照组,另外六组作为实验组,对照组不加入BDMC制剂,六个实验组分别一对一的加入浓度为5μM、10μM、15μM、20μM、25μM、30μM的BDMC制剂;以不同浓度的BDMC诱导hMSCs 6天后,终止培养。
2.观察结果:
如图2所示:采用BCIP/NBT染色法检测,细胞均成蓝紫色阳性反应,以15-25μM浓度条件下高表达,提示BDMC能诱导hMSCs生成成骨细胞的早期标志物的碱性磷酸酶,而对照组几乎不表达。
如图3A所示:通过酶活定量分析,呈一致结果,其中以20-25μM浓度条件下酶比活力最强。
如图3B所示:在5-30μM浓度范围内,BDMC对hMSCs无细胞毒性。
如图4所示,以20μM浓度的BDMC进一步分析其对成骨细胞标志物基因及蛋白的影响。BDMC诱导hMSCs 6天后,与对照组相比,诱导体系中的细胞高表达ALP、RUNX-2、OSX等成骨细胞分化早期相关基因(P< 0.05);诱导至21天时,与对照组相比,成骨细胞分化晚期相关基因COL1-α1、OCN、OPN的转录水平同样显著上调(P< 0.05)。
如图5所示:成骨细胞分化相关标志蛋白的检测结果呈一致的变化,即在诱导至第6天时,早期成骨细胞标志蛋白ALP、RUNX-2、OSX的表达水平明显高于对照组(P< 0.05);在诱导至第21天时,相比对照组,成骨细胞晚期标志蛋白COL1-α1、OCN、OPN的表达水平同样明显上调(P< 0.05)。而且,经BDMC诱导后的hMSCs不表达脂肪细胞相关标志物。
上述结果提示,hMSCs具有诱导hMSCs向成骨细胞定向分化的潜力。
如图6所示:进一步的细胞矿质化茜素红S染色检测分析表明,hMSCs经BDMC诱导至21天,能生成大量的成骨细胞特异性标志物红色钙化结节。
因此,BDMC(在体外)具有诱导hMSCs向成骨细胞定向分化的能力。
实施例3:BDMC联合hMSCs治疗骨质疏松症动物模型
1. 骨质疏松症小鼠动物模型构建:行卵巢切除术(OVX)构建骨质疏松症小鼠动物模型,术后恢复三周。
2. 实验分组设计与处理:将骨质疏松症小鼠动物模型分为6组,分别为:
对照组--简称NG,假性去除卵巢,不做任何处理;
模型组--简称MG,双侧去除卵巢,不做任何处理;
溶剂组--简称SG,双侧去除卵巢,每天用0.5%羧甲基纤维素钠经口灌胃给药处理;
BDMC药物治疗组--简称ivBG,双侧去除卵巢,按每天1次100 mg/kg剂量的BDMC经口灌胃给药;
hMSCs治疗组--简称hMSCs,双侧去除卵巢,每两周1次尾静脉输注移植2.0×106个hMSCs细胞;
BDMC与hMSCs联合治疗组--简称ivCG,双侧去除卵巢,按每天1次100 mg/kg剂量的BDMC经口灌胃给药,同时每两周1次尾静脉输注移植2.0×106 个hMSCs细胞。
hMSCs组与ivCG组中,hMSCs在尾静脉输注移植前用PBS缓冲液调配成hMSCs细胞浓度为2.0×106 个/200ul的人间充质干细胞制剂后再进行输注。
ivBG组与ivCG组中,灌胃给药前,先将BDMC充分溶解在0.5%羧甲基纤维素钠中制成BDMC制剂后才进行灌胃给药。
3. 结果分析
(1)骨量及骨微结构分析
骨质疏松症小鼠动物模型根据上述实验分组,连续治疗(处理)12周后,采用MicroCT分析不同治疗分组对骨质疏松症模型股骨骨量及骨微结构的影响。
结果如图7A所示,与对照组NG相比,模型组MG的骨量及骨微结构明显减少,具有典型的骨质疏松症状。经过不同治疗组治疗后,导致骨量及骨微结构有不同变化。溶剂对照组SG类似模型组MG,不能阻止骨量流失和骨微结构改变,表明药物助溶溶剂对实验结果无影响。而BDMC单独给药组ivBG和hMSCs细胞移植组均能一定程度阻止骨量流失和骨微结构改变,ivBG组甚至略好于hMSCs移植组。而BDMC与hMSCs联用组ivCG能更显著降低骨量丢失和减少骨微结构改变。进一步Micro CT影像数据定量分析呈一致结果,如图7B-7F所示,与对照组NG相比,MG组骨矿物质密度(BMD)、骨小梁数(Tb.N)、骨体积分数(BV/TV)均显著降低,而骨小梁分离度(Tb.Sp)显著增加(P< 0.01),溶剂对照SG组结果类似MG组。经过治疗后,相比MG组,3个不同治疗组的Tb.N明显增加,Tb.Sp明显减少(P< 0.05),但仅有ivBG组和ivCG组能明显增加BMD和BV/TV(P< 0.05),hMSCs组无明显逆转恢复效果。在这些组中,以联合治疗ivCG组效果最好,但各组间的骨小梁厚度(Tb.Th)无明显区别。因此,BDMC与hMSCs联合治疗具有防止骨质疏松症动物模型骨质流失和骨微结构改变的作用,而hMSCs单独移植治疗组并不能增加骨体积分数和骨矿物质密度,提示其单独使用不能提高骨质疏松症受体的成骨能力。
(2)骨吸收与骨形成相关标志物变化情况分析
基于骨质疏松症是骨代谢失衡,即破骨细胞的骨吸收水平高于成骨细胞的骨形成水平。为此进一步测定了各组血清中骨吸收与骨形成相关标志物变化情况。结果如图8所示,与对组组NG相比,模型组MG的骨吸收标志物CTX-1、TRAP的表达水平显著上调,而骨形成标志物OCN、PINP的表达水平显著降低(P< 0.01),溶剂对照组SG呈现与MG组一致结果,提示药物助溶溶剂对结果无影响。经过治疗后,相比MG组,ivBG和ivCG组均能明显降低CTX-1水平(P< 0.05),ivCG组也能显著降低TRAP水平,ivBG组对TRAP无明显影响,而hMSCs对CTX-1、TRAP两个骨吸收标志物均无改变。这3个不同治疗组均能促进骨形成标志物OCN的表达水平,但ivBG组无统计学意义。ivBG和ivCG组均能显著促进另一个骨形成标志物PINP的表达(P< 0.05),而hMSCs对PINP表达水平无影响。总之,BDMC和hMSCs联合组对骨质疏松症模型具有更显著的治疗效果,能明显抑制其骨吸收,促进其骨形成。
(3)肝脏毒性分析
基于目前骨质疏松症药物治疗的毒副作用,存在诸多安全性问题,我们进一步考察了这3种治疗方式对骨质疏松症模型的肝脏、肾脏等主要代谢器官的安全性。肝脏毒性结果如图9A所示,类似对照组NG的肝组织,ivBG组、hMSCs组、ivCG组等3个治疗组的肝组织结构清晰,肝细胞排列整齐,无明显的结构破坏和炎症细胞浸润等情况。但是,与对照组NG相比,骨质疏松症模型组MG所致的肝功能异常,使得谷草转氨酶(AST)和谷丙转氨酶(ALT)明显增加(如图9 B所示),溶剂组SG也呈一致结果,提示药物助溶溶剂对结果无影响。这3种治疗方式均能出现不同程度改善肝功能,降低AST/ALT水平,尤其是联合组ivCG具有显著的效果(P< 0.05),使其接近NG正常水平。因此,BDMC和hMSCs联合移植治疗具有明显逆转作用。
(4)肾脏毒性分析
肾脏毒性分析结果如图10A所示,ivBG组、hMSCs组、ivCG组等3个治疗组的肾皮质与髓质区域细胞结构清晰可见,排列整齐完整,未见细胞坏死、变性及结构异常细胞、炎症浸润等情况。但是,相比正常组NG,MG组血清中肌酐(Cr)及尿素氮(BUN)水平明显上调(如图10 B所示),经治疗后,3个治疗组的BUN出现轻微的下调,但对Cr水平有更好的改善作用,尤其是ivCG能明显降低Cr水平(P< 0.05)。因此,BDMC和hMSCs联合移植治疗对骨质疏松症模型肾功能有一定改善和逆转作用。
本发明的保护范围不限于具体实施方式所公开的技术方案,凡是依据本发明的技术实质对以上实施例所作的任何修改、等同替换、改进等,均落入本发明的保护范围。
Claims (7)
1. 天然化合物联合人间充质干细胞在制备治疗骨质疏松症药物中的应用,其特征在于:所述天然化合物为双去甲氧基姜黄素,分子式为C19H16O4, 化学结构式如通式(I)所示:
;
应用步骤为:将人间充质干细胞用PBS缓冲液调配成人间充质干细胞制剂,将双去甲氧基姜黄素溶解到0.5%羧甲基纤维素钠溶液中制成双去甲氧基姜黄素制剂,构建骨质疏松症哺乳动物模型;连续12周,每天用双去甲氧基姜黄素制剂对哺乳动物模型经口灌胃给药1次,同时每两周1次对哺乳动物模型尾静脉输注人间充质干细胞制剂。
2.根据权利要求1所述的应用,其特征在于:每两周1次对骨质疏松症哺乳动物模型尾静脉输注人间充质干细胞制剂的量为200ul;所述人间充质干细胞制剂中人间充质干细胞的浓度为2×106个/200μl;所述双去甲氧基姜黄素制剂中双去甲氧基姜黄素的使用剂量为100mg/kg/天。
3.根据权利要求2所述的应用,其特征在于:所述人间充质干细胞制剂选用对数生长期为2-5代的人间充质干细胞制成。
4.根据权利要求1-3中任意一项所述的应用,其特征在于:通过每天对骨质疏松症哺乳动物模型经口灌胃双去甲氧基姜黄素制剂来调节骨质疏松症动物模型体内微环境,在骨质疏松症体内营造利于人间充质干细胞向成骨细胞分化并限制向脂肪细胞分化的骨形成环境。
5.根据权利要求4所述的应用,其特征在于:所述骨质疏松症哺乳动物模型为骨质疏松症小鼠动物模型。
6.根据权利要求1所述的应用,其特征在于:所述天然化合物可与医学上可接受的药用辅料制成片剂、胶囊剂、丸剂、脂肪乳剂或软膏剂的药物。
7.根据权利要求6所述的应用,其特征在于:在制成的药物中所述天然化合物的使用剂量50-200mg/kg/天。
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