中国骨质疏松杂志

16S rDNA研究杜仲提取物5-HMF对去卵巢小鼠肠道菌群的影响

Study on the effect of 5-HMF extracted from Eucommia ulmoides on gut microbiota in ovariectomized mice using 16S rDNA

DOI：10.3969/j.issn.1006-7108.2024.12.004

中文关键词: 5-HMF 骨质疏松肠道菌群卵巢切除小鼠

英文关键词:5-HMF osteoporosis intestinal flora ovariectomized mice

基金项目:江苏省自然科学基金项目（BK20231147）；江苏省中医药管理局项目（MS2021044）；无锡市卫生健康委科研项目（Z202020）

作者	单位
刘小峰1 邵家豪2 何丹丹3 李超3 廖翼涛2 张贤3*	1.南通大学杏林学院附属惠山医院，无锡市惠山区人民医院，江苏无锡 214187 2.南京中医药大学，江苏南京 210023 3.南京中医药大学无锡附属医院，江苏无锡 214071

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中文摘要:

目的探讨从杜仲皮中提取出的生物活性化合物5-羟甲基糠醛（5-HMF）通过肠道菌群调控骨质疏松的作用机制。方法将24只8周龄雌性SD小鼠随机分为4组，其中18只小鼠建立双侧去卵巢骨质疏松模型后，将其随机分为去卵巢组（OVX组）、5-HMF给药组（5-HMF组）和戊酸雌二醇给药组（Positive组）；6只小鼠建立假手术组（Sham）组。将上述小鼠连续灌胃给药12周后，收集小鼠股骨和新鲜粪便。Micro-CT分析小鼠股骨骨量的变化；ELISA分析小鼠血清P1NP、E2、TNF-α、IL-6和IL-17水平；16S rDNA高通量测序检测小鼠粪便中肠道菌群变化情况。结果与Sham组比较，OVX组小鼠的股骨骨密度显著下降（P＜0.01），血清P1NP和E2水平显著下降（P＜0.001），血清TNF-α、IL-6和IL-17水平显著上升（P＜0.01），肠道菌群多样性显著增加（P＜0.01），菌群结构也发生了显著变化；与OVX组相比，5-HMF组小鼠股骨骨密度显著升高（P＜0.05），血清P1NP和E2水平显著上升（P＜0.01），血清TNF-α、IL-6和IL-17水平显著下降（P＜0.01），门水平上Bacteroidota菌群相对丰度升高，Firmicutes和Desulfobacterota菌群相对丰度显著下降，Firmicutes/Bacteroidota的相对丰度显著下降（P＜0.05）；属水平上Muribaculaceae和Alloprevotella的相对丰度升高，Lachnospiraceae_NK4A136_group的相对丰度降低。结论 5-HMF显著改善了去卵巢骨质疏松模型小鼠的骨密度、血清骨转换标志物和相关炎症因子水平，这可能与其调节小鼠肠道菌群结构相关。

英文摘要:

Objective To investigate the mechanism of 5-hydroxymethylfurfural (5-HMF), a bioactive compound extracted from the skin of Eucommia ulmoides, in regulating osteoporosis through intestinal flora. Methods 24 8-week-old female SD mice were randomly divided into 4 groups. 18 of them were randomly divided into OVX group, 5-HMF group and estradiol valerate group (Positive group) after bilateral ovariectomized osteoporosis model was established. Sham group was established in 6 mice. After 12 weeks of continuous intragastric administration of the above mice, the femur and fresh feces of the mice were collected. The changes of femoral bone mass in mice were analyzed by Micro-CT. Serum levels of P1NP, E2, TNF-α, IL-6 and IL-17 were analyzed by ELISA. 16S rDNA high-throughput sequencing was used to detect the changes of intestinal flora in mice faecal species. Results Compared with Sham group, bone mineral density of femur in OVX group was significantly decreased (P < 0.01), serum P1NP and E2 levels were significantly decreased (P < 0.001), serum TNF-α, IL-6 and IL-17 levels were significantly increased (P < 0.01), and intestinal flora diversity was significantly increased (P < 0.01). The bacterial community structure also changed significantly. Compared with OVX group, bone mineral density of femur in 5-HMF group was significantly increased (P < 0.05), serum levels of P1NP and E2 were significantly increased (P < 0.01), serum levels of TNF-α, IL-6 and IL-17 were significantly decreased (P < 0.01), and the relative abundance of Bacteroidota flora at the door level was increased. The relative abundance of Firmicutes and Desulfobacterota has decreased significantly, and the relative abundance of Firmicutes/Bacteroidota has decreased significantly (P < 0.05). At the genus level, the relative abundance of Muribaculaceae and Alloprevotella increased, while the relative abundance of Lachnospiraceae_NK4A136_group decreased. Conclusion 5-HMF can significantly improve the levels of bone mineral density, serum bone turnover markers and related inflammatory factors in ovariectomized osteoporosis mice, which may be related to its regulation of intestinal flora structure.

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