Galactooligosaccharides enhance intestinal barrier function and mitigate bone loss in ovariectomized mice

doi:10.3877/cma.j.issn.1674-0785.2025.01.008

Abstract

Abstract:

Objective

To investigate the effects of galactooligosaccharides （GOS） on bone metabolism in ovariectomized mice and the underlying mechanisms.

Methods

Eighteen C57BL/6J mice were randomly assigned to three groups： sham operation group （Sham）， ovariectomy-induced postmenopausal osteoporosis model group （OVX）， and treatment group （GOS）. Micro-CT was used to assess femoral microstructure， immunohistochemistry to evaluate osteogenesis and osteoclastogenesis， ELISA to measure serum levels of osteogenic and osteoclastic markers， 16S rRNA sequencing to analyze gut microbiota composition， H&E staining to examine intestinal barrier integrity， and Western blot and qPCR to detect expression levels of tight junction proteins and inflammatory factors.

Results

Compared with the Sham group， the bone mass in the OVX group was significantly reduced. In contrast， the GOS intervention group exhibited significant improvements in bone mass-related parameters （P＜0.01）. The OVX group showed reduced alkaline phosphatase （ALP） staining and increased tartrate-resistant acid phosphatase （TRAP）levels； GOS treatment significantly increased ALP and inhibited TRAP （P＜0.01）. Serum osteocalcin levels decreased while type I collagen C-terminal peptide levels increased in the OVX group； GOS treatment reversed these trends （P＜0.01）. 16S rRNA sequencing revealed that OVX reduced gut microbial diversity，whereas GOS reshaped the gut microbiota and increased the abundance of beneficial bacteria. H&E staining revealed that the OVX group exhibited impaired colonic intestinal barrier integrity， whereas GOS treatment significantly restored the intestinal structure. Western blot and qPCR analyses demonstrated that in the OVX group， the expression levels of tight junction proteins Claudin-1 and ZO-1 were downregulated， while the levels of the inflammatory cytokines TNF-α and IL-1β were upregulated. In contrast， GOS treatment increased the expression of Claudin-1 and ZO-1 and decreased the expression of TNF-α and IL-1β （P＜0.01）.

Conclusion

GOS mitigate bone loss in OVX mice by reshaping the intestinal microbiota and strengthening the intestinal barrier function.

Key words: Galactooligosaccharides, Postmenopausal osteoporosis, Gut microbiota, Intestinal barrier function

Peng Sheng, Lizhe1 Bai, Jing Zhang, Dan Li, Hong Cao. Galactooligosaccharides enhance intestinal barrier function and mitigate bone loss in ovariectomized mice[J]. Chinese Journal of Clinicians(Electronic Edition), 2025, 19(01): 48-57.

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References 43

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目的基因	上游引物	下游引物
β-actin	GTGACGTTGACATCCGTAAAGA	GCCGGACTCATCGTACTCC
Claudin-1	TCACTCCCAGGAGGATGC	GGCAGATCCAGTGCAAAGTC
ZO-1	CCCACCCCCATCTCAGAGTA	GGGACAGCTTTAGGCATGGT
RUNX2	GACTGTGGTTACCGTCATGGC	ACTTGGTTTTTCATAACAGCGGA
OCN	CTGACCTCACAGATCCCAAGC	TGGTCTGATAGCTCGTCACAAG
RANKL	AGCCGAGACTACGGCAAGTA	AAAGTACAGGAACAGAGCGATG
CTSK	CTCGGCGTTTAATTTGGGAGA	TCGAGAGGGAGGTATTCTGAGT
TNF-α	CAGGCGGTGCCTATGTCTC	CGATCACCCCGAAGTTCAGTAG
IL-1β	GAAATGCCACCTTTTGACAGTG	TGGATGCTCTCATCAGGACAG

目的基因	上游引物	下游引物
β-actin	GTGACGTTGACATCCGTAAAGA	GCCGGACTCATCGTACTCC
Claudin-1	TCACTCCCAGGAGGATGC	GGCAGATCCAGTGCAAAGTC
ZO-1	CCCACCCCCATCTCAGAGTA	GGGACAGCTTTAGGCATGGT
RUNX2	GACTGTGGTTACCGTCATGGC	ACTTGGTTTTTCATAACAGCGGA
OCN	CTGACCTCACAGATCCCAAGC	TGGTCTGATAGCTCGTCACAAG
RANKL	AGCCGAGACTACGGCAAGTA	AAAGTACAGGAACAGAGCGATG
CTSK	CTCGGCGTTTAATTTGGGAGA	TCGAGAGGGAGGTATTCTGAGT
TNF-α	CAGGCGGTGCCTATGTCTC	CGATCACCCCGAAGTTCAGTAG
IL-1β	GAAATGCCACCTTTTGACAGTG	TGGATGCTCTCATCAGGACAG

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