Effect of Zuoguiwan on mRNA Expression of Orexin and Its Receptors in Hypothalamus of Ovariectomized Rats

Su-min YE; Feng TAN; Fei-xiang LIU; Dao-ming XU; Yi CHAI; Ming-yue ZHANG; Qiao-ling FAN

doi:10.13422/j.cnki.syfjx.20191021

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Effect of Zuoguiwan on mRNA Expression of Orexin and Its Receptors in Hypothalamus of Ovariectomized Rats

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- Effect of Zuoguiwan on mRNA Expression of Orexin and Its Receptors in Hypothalamus of Ovariectomized Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 18, Pages: 9-15(2019)
- 作者机构：
  
  1.南京中医药大学　基础医学院，南京　 210029；
  2.江苏省中医院，南京　 210029
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191021
  CLC： R22; R242; R2-031; R285.5
- Published：20 September 2019，
  
  Published Online：13 February 2019，
  
  Received：09 September 2018，
- 稿件说明：
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Su-min YE, Feng TAN, Fei-xiang LIU, et al. Effect of Zuoguiwan on mRNA Expression of Orexin and Its Receptors in Hypothalamus of Ovariectomized Rats. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(18):9-15(2019)
DOI：

Su-min YE, Feng TAN, Fei-xiang LIU, et al. Effect of Zuoguiwan on mRNA Expression of Orexin and Its Receptors in Hypothalamus of Ovariectomized Rats. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(18):9-15(2019) DOI： 10.13422/j.cnki.syfjx.20191021.

摘要

目的：

观察去卵巢骨质疏松症大鼠下丘脑食欲素(Orexin)及其受体mRNA水平，探讨绝经后骨质疏松症(PMOP)的发病机制以及左归丸的作用机制。

方法：

去势法建立骨质疏松症模型，32只雌性SD大鼠随机分为假手术组、模型组、雌二醇组和左归丸组，每组8只。灌胃给药12周后，通过显微CT(

-CT)检测股骨骨密度(BMD)和骨小梁微观三维形态结构，苏木精-伊红(HE)染色观察大鼠骨组织形态学变化，酶联免疫吸附测定(ELISA)检测大鼠血清骨转换标志物骨钙素(OCN)，I型前胶原氨基端前肽(PINP)，抗酒石酸酸性磷酸酶(TRAP)含量，实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠下丘脑Orexin，食欲素受体1(OX1R)和食欲素受体2(OX2R)mRNA表达水平。

结果：

与假手术组比较，

-CT示模型组大鼠BMD，骨体积分数(BV/TV)，骨小梁厚度(Tb.Th)和骨小梁数量(Tb.N)显著下降(

0.01)而骨小梁间隙(Tb.Sp)升高(

0.01)，骨小梁明显稀疏，间距加宽，结构出现较大的空白区域；HE染色示骨小梁明显稀少，结构不完整，排列疏松，间距变宽；ELISA示血清OCN，PINP含量降低，TRAP含量升高(

0.01)；下丘脑Orexin，OX1R，OX2R mRNA表达下调(

0.01)。经左归丸治疗后，大鼠BMD，BV/TV，Tb.Th，Tb.N升高(

0.05，

0.01)，Tb.Sp降低(

0.01)，骨小梁变密，空隙率下降，分布均匀，排列尚可，微观结构明显改善；血清OCN，PINP含量增加(

0.05，

0.01)而TRAP含量降低(

0.01)；下丘脑Orexin，OX1R，OX2R mRNA表达明显上调(

0.05，

0.01)。

结论：

下丘脑Orexin及其受体mRNA水平降低可能是PMOP发病的机制之一。左归丸可通过上调下丘脑Orexin及其受体mRNA表达，从而纠正骨代谢失衡，改善骨小梁微结构，提高骨密度，发挥治疗PMOP的作用。

Abstract

Objective:

To observe the mRNA levels of Orexin and its receptors in the hypothalamus of ovariectomized osteoporosis rats

in order to explore the pathogenesis of postmenopausal osteoporosis(PMOP) and the mechanism Zuoguiwan.

Method:

An osteoporosis model induced by ovariectomy was established in rats. Totally 32 female Sprague-Dawley (SD) rats were randomly divided into sham-operated group

ovariectomized model group

-estradiol treated positive group

and Zuoguiwan group

with 8 rates in each group. After 12 weeks of intragastric administration

the bone mineral density (BMD) and trabecular microstructural changes of femur were detected by micro-CT (

-CT)

and the morphological changes of bone tissue were observed by hematoxylin-eosin staining (HE) staining. The markers of bone turnover in serum osteocalcin (OCN)

-terminal propeptide of type Ⅰ procollagen (PINP)

tartrate-resistant acid phosphatase (TRAP) were measured using enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of orexin

orexin receptor 1 (OX1R) and orexin receptor 2 (OX2R) were measured by Real-time PCR.

Result:

Compared with sham-operated group

the

-CT showed that BMD

bone volume fraction(BV/TV)

trabecular thickness(Tb. Th)and trabecular number(Tb. N)in ovariectomized model group were significantly decreased (

0.01)

while Trabecular Separation(Tb. Sp) was increased (

0.01)

and the bone trabeculae were obviously sparse

and the spacing was widened. HE staining showed that the trabecular bone was significantly rare

the structure was incomplete

the arrangement was loose

and the spacing was widened. ELISA showed that serum osteocalcin (OCN)

and

-terminal propeptide of type Ⅰ procollagen (PINP) levels decreased

whereas tartrate-resistant acid phosphatase (TRAP) content increased (

0.01)

and hypothalamic Orexin

OX1R

OX2R mRNA expressions were down-regulated (

0.01). After treatment

BMD

BV/TV

Tb. Th

Tb. N in Zuoguiwan group increased (

0.05

0.01)

while Tb. Sp decreased (

0.01)

bone trabeculae became dense

void fraction decreased

distribution was uniform

arrangement was acceptable

and the microstructure was significantly improved.serum OCN and PINP increased (

0.05

0.01)

while TRAP content decreased (

0.01)

the mRNA expressions of Orexin

OX1R and OX2R in the hypothalamus were significantly up-regulated (

0.05

0.01).

Conclusion:

Decreased mRNA levels of Orexin and its receptors in the hypothalamus may be one of the mechanisms of PMOP. Zuoguiwan may correct the imbalance of bone metabolism

improve the trabecular microstructure and improve bone by up-regulating the mRNA expressions of Orexin and its receptors in the hypothalamus

density

so as to show a therapeutic effect on PMOP.

关键词

左归丸绝经后骨质疏松症食欲素食欲素受体1食欲素受体2

Keywords

Zuoguiwanpostmenopausal osteoporosis(PMOP)orexinorexin receptor 1 (OX1R)orexin receptor 2 (OX2R)

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