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陈一洲,叶勇健,李之斌,陈丹琦.金匮肾气丸调控β-catenin通路治疗去卵巢小鼠骨质疏松[J].浙江中西医结合杂志,2020,30(10):
金匮肾气丸调控β-catenin通路治疗去卵巢小鼠骨质疏松
Kidney-qi-tonifying pills Attenuate Osteoporosis via regulation of β-catenin Signaling Pathway in Ovariectomized Mice
投稿时间:2020-04-06  修订日期:2020-08-19
DOI:
中文关键词:  金匮肾气丸  骨质疏松症  β-catenin  去卵巢小鼠
英文关键词:kidney-qi-tonifying pills  osteoporosis  β-catenin  ovariectomized mice
基金项目:
作者单位E-mail
陈一洲 宁波市鄞州区第二医院 103486524@qq.com 
叶勇健 宁波市鄞州区第二医院  
李之斌 宁波市鄞州区第二医院  
陈丹琦 宁波市鄞州区第二医院 121451449@qq.com 
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中文摘要:
      目的:探究金匮肾气丸治疗去卵巢小鼠骨质疏松的疗效和分子机制。方法:24只10周龄雌性C57BL/6小鼠按照随机数字表法分为假手术组、模型组和金匮肾气丸组,每组8只。模型组和金匮肾气丸组小鼠摘除双侧卵巢造模,假手术组切除卵巢周围等量的脂肪组织。术后第4天开始灌胃干预,金匮肾气丸组给予金匮肾气丸溶液0.2 mL/天,模型组给予等量生理盐水。干预8周后,摘取小鼠双侧股骨进行Micro-CT、病理染色和qRT-PCR检测。结果:与假手术组比较,模型组骨密度[(0.90±0.07)mg/mm3比(1.21±0.10)mg/mm3;P<0.05]、骨体积分数[(26.31±2.90)%比(42.10±5.35)%;P<0.05]和骨小梁数目[(2.39±0.29)mm比(3.51±0.42)mm;P<0.05]均降低,骨小梁间距增加[(0.34±0.08)mm比(0.21±0.05)mm;P<0.05],骨小梁厚度无显著变化[(0.12±0.01)mm比(0.12±0.03)mm;P>0.05],骨小梁面积分数减少(12.89±4.36)%比(22.52±2.32)%;P<0.05],骨髓腔内脂肪空泡面积[(13.01±4.95)%比(3.03±0.66)%;P<0.05]和直径[(32.03±5.62)μm比(23.22±1.90)μm;P<0.05]均增加。β-catenin及其下游Osterix和ALP基因表达减少[β-catenin:(0.387±0.139)比(1.036±0.337);Osterix:(0.362±0.134)比(1.114±0.350);ALP:(0.272±0.077)比(1.004±0.118);P<0.05)]。经金匮肾气丸治疗后,小鼠股骨骨密度升高[(1.15±0.12)mg/mm3比(0.90±0.07)mg/mm3;P<0.05],骨体积分数[(34.24±4.01)%比(26.31±2.90)%;P<0.05]和骨小梁数目[(2.72±0.25)mm比(2.39±0.29)mm;P<0.05]均升高,骨小梁间距减少[(0.26±0.03)mm比(0.34±0.08)mm;P<0.05],骨小梁厚度无显著变化[(0.12±0.02)mm比(0.12±0.01)mm;P>0.05],骨小梁面积分数增加[(20.17±4.88)%比(12.89±4.36)%;P<0.05],髓腔内脂肪空泡面积[(6.96±3.58)%比(13.01±4.95)%;P<0.05]和直径[(25.43±3.91)μm比(32.03±5.62)μm;P<0.05]均减少。β-catenin、Osterix和ALP的mRNA表达升高[β-catenin:(0.877±0.224)比(0.387±0.139);Osterix:(0.881±0.187)比(0.362±0.134);ALP:(0.812±0.145)比(0.272±0.077);P<0.05]。结论:金匮肾气丸能够有效延缓去卵巢小鼠骨量丢失,其可能是促进β-catenin及下游Osterix、ALP成骨基因表达,进而影响干细胞向成骨细胞分化。
英文摘要:
      Objective: To investigate the effects and molecular mechanism of kidney-qi-tonifying pills on osteoporosis in ovariectomized mice. Methods: Twenty-four 10-weeks old female C57/BL6 mice were randomly divided into the sham-operation group, the model group and the kidney-qi-tonifying pills group by random number table method. Mice in the model and the kidney-qi-tonifying pills groups were received a resection of bilateral ovaries, while mice in the sham-operation group were only resected the equivalent para-ovarian adipose tissue. Since 4 days post-surgery, mice in the kidney-qi-tonifying pills group began to receive 0.2 ml/day drug lavage intervention, and mice in the model group receive normal saline in the same volume. At 8 weeks after the intervention, bilateral femurs were harvested from the experimental mice for Micro-CT analysis, pathological staining and qRT-PCR analysis. Results: Compared with the sham operation group, mice in the model group presented low bone mineral density [(0.90±0.07) mg/mm3 vs (1.21±0.10) mg/mm3; P<0.05], decreased bone volume fraction [(26.31±2.90) % vs (42.10±5.35) %; P<0.05], decreased trabecular number [(2.39±0.29) mm vs (3.51±0.42) mm; P<0.05], increased trabecular separation [(0.34±0.08) mm vs (0.21±0.05) mm; P<0.05], unchanged trabecular thickness [(0.12±0.01) mm vs (0.12±0.03) mm; P>0.05], decreased bone area fraction [(12.89±4.36) % vs (22.52±2.32) %; P<0.05], and increased lipid droplet area [(13.01±4.95) % vs (3.03±0.66) %; P<0.05] and diameter [(32.03±5.62) μm vs (23.22±1.90) μm; P<0.05] as well as decreased mRNA expression of β-catenin, Osterix and alkaline phosphatase (ALP) [β-catenin: (0.387±0.139) vs (1.036±0.337); Osterix: (0.362±0.134) vs (1.114±0.350); ALP: (0.272±0.077) vs (1.004±0.118); P<0.05]. After the treatment of kidney-qi-tonifying pills, mice showed improvement of bone mineral density [(1.15±0.12) mg/mm3 vs (0.90±0.07) mg/mm3; P<0.05], bone volume fraction [(34.24±4.01) % vs (26.31±2.90) %; P<0.05] and trabecular number [(2.72±0.25) mm vs (2.39±0.29) mm; P<0.05], decrease of trabecular separation [(0.26±0.03) mm vs (0.34±0.08) mm; P<0.05], unchanged trabecular thickness [(0.12±0.02) mm vs (0.12±0.01) mm; P>0.05], increased bone area fraction [(20.17±4.88) % vs (12.89±4.36) %; P<0.05], decreased lipid droplet area [(6.96±3.58) % vs (13.01±4.95) %; P<0.05] and diameter [(25.43±3.91) μm vs (32.03±5.62) μm; P<0.05], as well as increased mRNA levels of β-catenin, Osterix and ALP [β-catenin: (0.877±0.224) vs (0.387±0.139); Osterix: (0.881±0.187) vs (0.362±0.134); ALP: (0.812±0.145) vs (0.272±0.077); P<0.05]. Conclusion: Kidney-qi-tonifying pills could attenuate bone loss in ovariectomized mice possibly through activation of β-catenin and downstream osteogenic target gene (Osterix and ALP), which might promote osteogenic differentiation of mesenchyml stem cells.
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