Bone-Protective Role of Bupleurum in a Rat Model of Postmenopausal Osteoporosis

Summary

In this study, we used an ovariectomized rat model of postmenopausal osteoporosis to demonstrate that Bupleurum (Chai hu) oral liquid improves bone microstructure by suppressing pro-inflammatory factors and enhancing anti-inflammatory ones. This suggests that its anti-inflammatory mechanism partially mitigates estrogen deficiency-induced osteoporosis.

Abstract

The protocol aims to investigate the effects of Chai hu administration on postmenopausal osteoporosis in ovariectomized (OVX) rats. Eight-week-old female Sprague-Dawley rats underwent bilateral ovariectomy using the dorsal approach to establish a postmenopausal osteoporosis model. Different doses of Chai hu oral liquid were administered by oral gavage for 12 consecutive weeks, followed by evaluations of bone microstructure and measurements of inflammatory cytokine levels. Medium- and high-dose Chai hu treatment partially inhibited bone loss, reduced pro-inflammatory cytokine expression, and upregulated anti-inflammatory cytokine levels in OVX rats. There was a significant reduction in the severity of bone destruction from low to high doses, and dose-related improvements in bone microstructural reconstruction were observed. Chai hu mitigated estrogen deficiency-induced bone loss and microstructural deterioration, thereby suppressing OP progression in this OVX model. This was likely to have resulted from the anti-inflammatory effects of the pathways involved.

Introduction

Osteoporosis (OP) is a common metabolic bone disease characterized by decreased bone mass, reduced bone strength, and destruction of bone microarchitecture¹. OP can manifest as either primary or secondary, and primary OP is classified as postmenopausal OP (PMOP), senile OP, and idiopathic OP (which often occurs in adolescents). Among them, type I PMOP occurs primarily in PM women, where there is a marked decrease in bone density after menopause due to reduced circulating levels of estrogen². It is a systemic metabolic bone disease, which results in increased osteoclast activity and intense differentiation

Protocol

All experimental procedures involving animals were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals (8th edition) and were specifically approved by the Animal Ethics Committee of Youjiang Medical University for Nationalities (Approval No. 2024071001).

1. Analyzing network pharmacology

NOTE: Cyberpharmacology is an approach that combines systems biology, computational biology, and pharmacology and aims to understand the mechanism of action of drugs and drug-drug interactions by analyzing biomolecular networks²⁵.

1. Screen active....

Results

GO function and KEGG pathway enrichment analysis

We relied on GO and KEGG enrichment analysis in bioinformatics to predict the potential mechanism of action of Chai Hu on PMOP. The GO enrichment analysis of target genes intersecting with the active ingredients of Chai hu for diseases yielded 2138 significant entries, and the top ten significant results are listed for each group (Figure 2D). With respect to biologi.......

Discussion

PMOP is a systemic disorder of bone metabolism that often occurs in PM women, and it is caused by a lack of estrogen that results in a disruption of the osteoblast-osteoclast balance, leading to enhanced osteoclast bone resorption and diminished osteoblast bone production³¹. A growing number of studies have shown that the levels of inflammatory factors in the body also affect the differentiation between osteoclasts and osteoblasts, and that overexpression of pro-inflammatory factors contributes to.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.9% sodium chloride injection	Huarenpharmaceutical	h1712059
1×PBS	Thermofisher	10010023
75% alcohol	Nanjingreagent	C069156931
95% ethanol	Nanjingreagent	C0691520075
Anhydrous ethanol	Macklin	E809056
Bupleurum chinense	Henan Baiquan Pharmacy Co.	Z41021539
decalcification solution	solarbio	E1171
erythromycin ointment	Guangzhou Baiyunshan Pharmaceutical Group Company Limited Baiyunshan Hejigong Pharmaceutical Factory	H44023088
Isopropyl alcohol	Jinan Jinrihe Chemical Co.	23727-29-9
penicillin V potassium capsules	Sichuan Pharmaceutical	H20103156
polyformaldehyde (CH2O)n	Beyotime	P0099
povidone-iodine disinfectant	Guangdong Hengjian Pharmaceutical Co.	H44023924
Real-Time Fluorescence PCR Kit	Yeasen Biotechnology	11201ES03
RNA Extraction Kit	Monadbiotech	MI20201S
RNA Reverse Transcription Kit	YuGongBiotech	EG15123S
Sodium pentobarbital	Shanghai Jixiang Biotechnology Co.	JX082085
Trizol	Thermofisher	15596026CN