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[牙周炎对雌激素缺乏小鼠肠道微生物群的骨代谢和色氨酸代谢的影响]

[Effects of periodontitis on bone and tryptophan metabolism of gut microbiota in estrogen-deficient mice].

作者信息

Wei Y, Wang N N, Wang M, Qian J, Nie H, Ge R Y, Liao W Z, Yan F H

机构信息

Department of Periodontology, Nanjing Stomatological Hospital, Affiliated Hospital of Medical School,Institute of Stomatology, Nanjing University, Nanjing 210008, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2024 Apr 9;59(4):354-363. doi: 10.3760/cma.j.cn112144-20231225-00307.

Abstract

To study the effects of periodontitis on bone and tryptophan metabolism of gut microbiota in the context of estrogen deficiency. Thirty-two female C57BL6/J mice were randomly divided into four groups based on table of random numbers (8 in each group): Sham group, in which mice were given sham surgery; Sham_Lig group, in which mice were given sham surgery and were induced to periodontitis by ligating the bilateral maxillary second molars with 5-0 silk threads at the fourth week; Ovx group, in which mice were given bilateral ovariectomy; Ovx_Lig group, in which mice were given bilateral ovariectomy and were induced to periodontitis at the fourth week. After 8 weeks of ligation, the mice of 4 groups were euthanized for collecting the samples of femur, tibia, mandible and skull. Those samples were scanned by micro-CT to measure the bone mineral density (BMD), bone volume versus total volume ratio (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th) and trabecular spacing (Tb.Sp). The cecum contents of 4 groups of mice were collected for gut microbiota 16S rRNA gene sequencing. The tryptophan and its metabolites in intestinal tracts were detected by liquid chromatography-mass spectrometry. Pearson correlation analysis was performed to analyze the correlation between the abundance of gut microbiota and the content of tryptophan and its metabolites. Femur BMD [(82.23±3.97) mg/cm], BV/TV [(9.25±1.37)%] and Tb.Th [(70.95±5.70) μm] in Ovx_Lig group were significantly lower than Ovx group [(96.30±3.76) mg/cm (0.004); (14.45±1.55)% (0.022) and (87.58±8.02) μm (0.001), respectively]. The β-diversity analysis of gut microbiota based on Bray-Curtis distance showed that samples of Ovx_Lig group and Ovx group were obviously grouped. Linear discriminant analysis effect size (LEfSe) showed that was the representative genus in Ovx_Lig group. The relative abundance of in Ovx_Lig group [(0.42±0.14)%] were significantly higher than that in Ovx group [(0.17±0.05)%] (4.45, 0.001). Tryptophan metabolism analysis showed that the content of kynurenic acid [(531.12±158.60) ng/g] in Ovx_Lig group were significantly higher than that in Ovx group [(400.42±57.96) ng/g] (2.19, 0.046). And the content of indole-3-carbaldehyde [(383.37±144.06) ng/g] in Ovx_Lig group were significantly lower than Ovx group [(701.72±141.93) ng/g] (4.45, 0.001). Correlation analysis showed that relative abundance of was positively correlated with kynurenic acid (0.32, 0.088), while negatively correlated with indole-3-carbaldehyde (-0.32, 0.088). Periodontitis can induce bone destruction of femur in estrogen-deficient mice, the mechanism of which may be related to in gut and the tryptophan metabolites kynurenic acid and indole-3-carbaldehyde.

摘要

研究雌激素缺乏背景下牙周炎对肠道微生物群骨代谢和色氨酸代谢的影响。将32只雌性C57BL6/J小鼠根据随机数字表随机分为四组(每组8只):假手术组,小鼠接受假手术;假手术结扎组,小鼠接受假手术,并在第4周用5-0丝线结扎双侧上颌第二磨牙诱导牙周炎;去卵巢组,小鼠接受双侧卵巢切除术;去卵巢结扎组,小鼠接受双侧卵巢切除术,并在第4周诱导牙周炎。结扎8周后,对4组小鼠实施安乐死以收集股骨、胫骨、下颌骨和颅骨样本。对这些样本进行显微CT扫描以测量骨密度(BMD)、骨体积与总体积比(BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)和骨小梁间距(Tb.Sp)。收集4组小鼠的盲肠内容物进行肠道微生物群16S rRNA基因测序。采用液相色谱-质谱法检测肠道中的色氨酸及其代谢产物。进行Pearson相关性分析以分析肠道微生物群丰度与色氨酸及其代谢产物含量之间的相关性。去卵巢结扎组的股骨BMD[(82.23±3.97)mg/cm]、BV/TV[(9.25±1.37)%]和Tb.Th[(70.95±5.70)μm]显著低于去卵巢组[(96.30±3.76)mg/cm(P = 0.004);(14.45±1.55)%(P = 0.022)和(87.58±8.02)μm(P = 0.001)]。基于Bray-Curtis距离的肠道微生物群β多样性分析表明,去卵巢结扎组和去卵巢组的样本明显聚类。线性判别分析效应大小(LEfSe)显示,[具体属名未给出]是去卵巢结扎组的代表性属。去卵巢结扎组中[该属名]的相对丰度[(0.42±0.14)%]显著高于去卵巢组[(0.17±0.05)%](P = 4.45,0.001)。色氨酸代谢分析表明,去卵巢结扎组中犬尿酸的含量[(531.12±158.60)ng/g]显著高于去卵巢组[(400.42±57.96)ng/g](P = 2.19,0.046)。而去卵巢结扎组中吲哚-3-甲醛的含量[(383.37±144.06)ng/g]显著低于去卵巢组[(701.72±141.93)ng/g](P = 4.45,0.001)。相关性分析表明,[该属名]的相对丰度与犬尿酸呈正相关(r = 0.32,P = 0.088),而与吲哚-3-甲醛呈负相关(r = -0.32,P = 0.088)。牙周炎可诱导雌激素缺乏小鼠股骨的骨质破坏,其机制可能与肠道中的[具体属名未给出]以及色氨酸代谢产物犬尿酸和吲哚-3-甲醛有关。

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