Effects and mechanism of human umbilical cord mesenchymal stem cells on osteoporosis rats by regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-κB ligand

Ying Wei; Shiwei Liu; Ruixue Duan; Yuxiang Zhao; Nan Li; Yaru Wu

doi:10.13406/j.cnki.cyxb.002972

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Effects and mechanism of human umbilical cord mesenchymal stem cells on osteoporosis rats by regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-κB ligand

Orthopedics | 更新时间：2022-06-24

- Effects and mechanism of human umbilical cord mesenchymal stem cells on osteoporosis rats by regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-κB ligand
- Journal of Shanxi Agricultural Sciences Vol. 47, Issue 5, Pages: 503-510(2022)
- 作者机构：
  
  1.山西医科大学基础医学院生物化学与分子生物学教研室，太原　030001
  2.山西白求恩医院内分泌科，太原　030032
  3.太原市中心医院内分泌科，太原　030009
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.002972
  CLC： R580
- Published：28 May 2022，
  
  Received：15 May 2021，
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Ying Wei, Shiwei Liu, Ruixue Duan, et al. Effects and mechanism of human umbilical cord mesenchymal stem cells on osteoporosis rats by regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-κB ligand. [J]. Journal of Shanxi Agricultural Sciences 47(5):503-510(2022)
DOI：

Ying Wei, Shiwei Liu, Ruixue Duan, et al. Effects and mechanism of human umbilical cord mesenchymal stem cells on osteoporosis rats by regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-κB ligand. [J]. Journal of Shanxi Agricultural Sciences 47(5):503-510(2022) DOI： 10.13406/j.cnki.cyxb.002972.

摘要

目的：

探讨人脐带间充质干细胞（human umbilical cord mesenchymal stem cells，HUC-MSCs）对骨质疏松大鼠的影响及机制。

方法：

将30只SPF级6月龄雌性SD大鼠随机分为3组：假手术组（Sham组）、卵巢切除组（OVX组）、卵巢切除+HUC-MSCs干预组（4×10

/3 d，HUC-MSCs干预组），每组各10只，Sham组切除大鼠腹腔少量脂肪，其余各组均去除双侧卵巢建立骨质疏松大鼠模型。建模成功后，HUC-MSCs组用干细胞干预12周，其余2组用等剂量生理盐水干预。干预结束后，通过骨代谢血清指标血钙（calcium，Ca）、血磷（phosphorus，P）、碱性磷酸酶（alkaline phosphatase，ALP）、Ⅰ型胶原C端肽（c-terminal peptide of typeⅠcollagen，CTX）、骨组织形态计量学参数测量、骨密度检测、成骨细胞表面积/骨表面积比（osteoblast surface/bone surface，ObS/BS）和破骨细胞表面积/骨表面积比（osteoclast surface area/bone surface，OcS/BS），评估大鼠骨质疏松改善情况，同时通过Western blot探究各组大鼠骨保护素（osteoprotegerin，OPG）/核因子-κB受体活化因子配体（receptor activator of nuclear factor-κB ligand，RANKL）、β-连环蛋白（β-catenin）、Runt相关转录因子2（Runt-related transcription factor 2，Runx2）的表达情况。

结果：

HUC-MSCs组大鼠血清Ca、ALP含量明显高于OVX组（2.320±0.188 vs.1.822±0.139；79.584±8.349 vs.43.644±5.369；均

=0.000），P、CTX含量明显低于OVX组（1.651±0.068 vs.1.864±0.076，

=0.000；119.832±18.180 vs.144.272±15.629，

=0.039）；HUC-MSCs组骨小梁结构得到明显改善，骨密度、骨小梁数目、骨小梁面积比、ObS/BS均明显高于OVX组（161.114±16.726 vs.126.558±7.100，

=0.009；1.702±0.220 vs.1.051±0.323，

=0.020；33.111±6.101 vs.19.575±5.629，

=0.022；45.279±3.947 vs.33.948±7.339，

=0.041），骨小梁分离度、OcS/BS明显低于OVX组（0.592±0.066 vs.0.845±0.165，

=0.018；37.532±3.910 vs.44.891±3.615，

=0.026）；Western blot结果显示HUC-MSCs组OPG/RANKL比值、β-catenin、Runx2的蛋白表达量明显高于OVX组（0.776±0.045 vs.0.097±0.063，

=0.000；0.590±0.057 vs.0.202±0.088，

=0.000；0.595±0.078 vs.0.239±0.071，

=0.001）。

结论：

HUC-MSCs可以通过Wnt/β-catenin/Runx2通路调节OPG/RANKL比值，促进骨形成，抑制骨吸收，改善骨质疏松。

Abstract

Objective

To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on osteoporosis rats and its mechanism.

Methods

Thirty SPF grade 6-month-old female rats were randomly divided into three groups: sham operation group (Sham group)

ovariectomy group (OVX group) and HUC-MSCs intervention group (HUC-MSCs group) (4×10

/3 day)

with 10 rats in each group. In Sham group

a small amount of abdominal fat was removed

and the other groups were removed bilateral ovaries to establish osteoporosis rat model. After successful modeling

HUC-MSCs group was intervened with stem cells for 12 w

while the other two groups were intervened with equal dose of normal saline. After the intervention

calcium (Ca)

phosphorus (P)

alkaline phosphatase (ALP)

c-terminal peptide of type I collagen (CTX)

histomorphometric parameters of femur

bone mineral density (BMD)

osteoblast surface/bone surface ratio (ObS/BS) and osteoclast surface area/bone surface (OcS/BS) were used to evaluate the improvement of osteoporosis in rats. Meanwhile

the expression of osteoprotegerin (OPG)

receptor activator of nuclear factor-κB ligand (RANKL)

β-catenin and Runt-related transcription factor 2 (Runx2) were detected by Western blot.

Results

The contents of serum Ca and ALP in HUC-MSCs group were significantly higher than those in OVX group (2.320±0.188 vs.1.822±0.139; 79.584±8.349 vs.43.644±5.369; all

=0.000) ; P and CTX contents were significantly lower than those in OVX group (1.651±0.068 vs.1.864±0.076

=0.000; 119.832±18.180 vs.144.272±15.629

=0.039) ; BMD

the number of trabecular bone

the ratio of trabecular area and ObS/BS in HUC-MSCs group were significantly higher than those in OVX group (161.114±16.726 vs.126.558±7.100

=0.009; 1.702±0.220 vs.1.051±0.323

=0.020; 33.111±6.101 vs.19.575±5.629

=0.022; 45.279±3.947 vs.33.948±7.339

=0.041) ; the bone trabecular separation and OcS/BS in HUC-MSCs group were significantly lower than those in OVX group (0.592±0.066 vs.0.845±0.165

=0.018; 37.532±3.910 vs.44.891±3.615

=0.026) ; the results of Western blot showed that the ratio of OPG/RANKL and the protein expression of β-catenin and Runx2 in HUC-MSCs group were significantly higher than those in OVX group (0.776±0.045 vs.0.097±0.063

=0.000; 0.590±0.057 vs.0.202±0.088

=0.000; 0.595±0.078 vs.0.239±0.071

=0.001) .

Conclusion

HUC-MSCs can regulate OPG/RANKL ratio through Wnt/β-catenin/Runx2 pathway

thus promoting bone formation

inhibiting bone resorption and improving osteoporosis.

关键词

骨质疏松人脐带间充质干细胞骨代谢骨保护素/核因子-κB受体活化因子配体

Keywords

osteoporosishuman umbilical cord mesenchymal stem cellbone metabolismosteoprotegerin/receptor activator of nuclear factor-κB ligand

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