标题：齐墩果酸经肠道菌群抑制去卵巢小鼠骨丢失的作用机制

      作者：马承红 1，王姜俨 1，苏丹丹 2，谢佑红 1，唐琳 1，周天宇 1，董群伟 3，4，孙平 1    马承红 1，王姜俨 1，苏丹丹 2，谢佑红 1，唐琳 1，周天宇 1，董群伟 3，4，孙平 11.广东药科大学附属第一医院内分泌科，广东 广州 510062；2.广东药科大学附属第一医院信息科，广东 广州 510062；3.广东药科大学附属第一医院骨科，广东 广州 510062；4.广东省云浮市中医院，广东 云浮 527300

      卷次： 2023年34卷22期

      【摘要】 目的 基于肠道菌群的变化探讨齐墩果酸(OA)防治去卵巢(OVX)小鼠骨丢失机制。方法 19只11周龄C57BL6/J雌性小鼠随机分为假手术组(SHAM组，n=6)、OVX组(n=6)和OA组(n=7)。OA组小鼠在OVX基础上腹腔注射OA (10 mg·kg-1·2 d-1)，SHAM组和OVX组小鼠则腹腔注射等剂量生理盐水，12周后收集小鼠左侧股骨、血清和粪便标本。Micro-CT检测小鼠左侧股骨骨密度(BMD)、骨体积分数(BV/TV)、骨小梁数量(Tb.N)及骨小梁分离度(Tb.Sp)等；ELISA法检测小鼠血清中1型前胶原氨基端前肽(P1NP)、β-胶原降解产物(β-CTX)、抗酒石酸酸性磷酸酶 5b (TRACP5b)、肿瘤坏死因子-α (TNF-α)、白细胞介素-10 (IL-10)、白细胞介素-6 (IL-6)及脂多糖(LPS)水平；16S rRNA高通量肠菌基因测试检测小鼠肠道菌群相关指标。结果 OVX组小鼠的BMD、BV/TV、Tb.N和 IL-10较SHAM组明显降低，Tb.Sp、P1NP、β-CTX、TRACP5b、TNF-α和 IL-6明显增加，差异均有统计学意义(P<0.05或<0.01)；OA组小鼠的BMD、BV/TV和 IL-10较OVX组明显增加，Tb.Sp、P1NP、β-CTX、TRACP5b、TNF-α和IL-6明显降低，差异均具有统计学意义(P<0.05或<0.01)，但Tb.N变化不明显，差异无统计学意义(P>0.05)。OVX组小鼠的放线菌丰度较SHAM组明显降低，OA组小鼠放线菌丰度较OVX组明显增高，差异均具有统计学意义(P<0.01)。在属水平，与SHAM组相比，葡萄球菌、腔肠杆菌科_UCG-002和肠杆菌丰度在OVX组中明显降低，臭杆菌丰度明显增高，差异均具有统计学意义(P<0.01或<0.05)；与OVX组相比，OA组小鼠的葡萄球菌、腔肠杆菌科_UCG-002和肠杆菌丰度明显增高，臭杆菌丰度明显降低，差异均具有统计学意义(P<0.01或<0.05)。结论 OA可延缓OVX小鼠骨丢失、增加骨量，其机制可能是通过降低炎性反应、改善肠道通透性及调节肠道菌群结构实现的。
      【关键词】 去卵巢小鼠；齐墩果酸；骨质疏松；肠道菌群；骨丢失；作用机制
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2023）22—3193—07

Role and mechanism of oleanolic acid in inhibiting bone loss of ovariectomized mice via gut microbiota.
MACheng-hong 1, WANG Jiang-yan 1, SU Dan-dan 2, XIE You-hong 1, TANG Lin 1, ZHOU Tian-yu 1, DONG Qun-wei 3,4,SUN Ping 1. 1. Department of Endocrinology, the First Affiliated Hospital of Guangdong Pharmaceutical University,Guangzhou 510062, Guangdong, CHINA; 2. Department of Information, the First Affiliated Hospital of GuangdongPharmaceutical University, Guangzhou 510062, Guangdong, CHINA; 3. Department of Orthopedics, the First AffiliatedHospital of Guangdong Pharmaceutical University, Guangzhou 510062, Guangdong, CHINA; 4. Yunfu Hospital ofTraditional Chinese Medicine, Yunfu 527300, Guangdong, CHINA
【Abstract】 Objective To explore the mechanism of oleanolic acid (OA) against bone loss in ovariectomized(OVX) mice based on the alteration of gut microbiota. Methods Nineteen 11-week-old C57BL6/J female mice weredivided into three groups randomly: SHAM group (n=6), OVX group (n=6), and OA group (n=7). The mice in OA groupwere injected intraperitoneally with OA (10 mg·kg-1·2 d-1) on the basis of OVX, and the mice in SHAM and OVXgroups were injected intraperitoneally an equal dose of 0.9% NaCl. After 12 weeks, the left femurs, serum, and fecalspecimens were collected. Micro-CT was used to detect bone mineral density (BMD), bone volume fraction (BV/TV),trabecular number (Tb.N), and trabecular spacing (Tb.Sp) of the left femurs of the mice. The levels of procollagen type 1N-terminal propeptide (P1NP), β-collagen degradation products ( β-CTX), tartrate resistant acid phosphatase 5b(TRACP5b), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), interleukin-6 (IL-6), and lipopolysaccharides(LPS) in serum of mice were detected by ELISA. 16S rRNA sequencing was used to detect the related indexes of gut mi-crobiota in mice. Results BMD, BV/TV, Tb.N, and IL-10 in the OVX group were significantly lower than those in theSHAM group, while Tb.Sp, P1NP, β-CTX, TRACP5b, TNF-α, and IL-6 were significantly higher, with statistically sig-nificant differences (P<0.05 or P<0.01). BMD, BV/TV, and IL-10 in the OA group were significantly higher than thosein the OVX group, while Tb.Sp, P1NP, β-CTX, TRACP5b, TNF-α, and IL-6 were significantly lower than those in theOVX group, with statistically significant differences (P<0.05 or <0.01). There was no significant change in Tb.N be-tween the OA group and OVX group (P>0.05). At the phylum level, the abundance of Actinobacteriota in the OVXgroup was significantly lower than that in the SHAM group, and the abundance of Actinobacteriota in the OA group wassignificantly higher than that in the OVX group, with statistically significant differences (P<0.01). At the genus level,compared with the SHAM group, the abundance of Staphylococcus, Coriobacteriaceae_UCG-002, and Enterorhabduswere significantly decreased in the OVX group, and the abundance of Odoribacter was significantly increased, with sta-tistically significant differences (P<0.01 or P<0.05). Compared with the OVX group, the abundance of Staphylococcus,Coriobacteriaceae_UCG-002, and Enterorhabdus in the OA group were significantly increased, and the abundance ofOdoribacter was significantly decreased, with statistically significant differences (P<0.01 or P<0.05). Conclusion OAcan reduce bone loss and increase bone mass in OVX mice. Its role may be achieved by relieving inflammation, improv-ing intestinal permeability, and regulating gut microbiota structure.
      【Key words】 Ovariectomized mice; Oleanolic acid; Osteoporosis; Gut microbiota; Bone loss; Mechanism