Effect of Bifidobacterium trisporus on Intestinal Microbiota in the Mouse Model of Non-alcoholic Fatty Liver Disease
-
摘要:
目的 探究双歧杆菌三联活菌在小鼠NAFLD模型中的疗效和对肠道微生物组的影响。 方法 使用C57BL/6J小鼠分为3个实验组别，对照组（n = 10）：小鼠使用普通饮食饲养；NAFLD模型组（n = 10）：使用高脂饮食饲养；双歧杆菌组（n = 10）：使用NAFLD模型小鼠并给予双歧杆菌三联活菌治疗。在实验结束时间点对不同组小鼠肝脏石蜡切片进行HE染色观察病理变化。采用16S rRNA基因测序技术完成对各组小鼠肠道菌群的检测，并进行生物信息学分析。 结果 与对照组相比，NAFLD小鼠肝脏脂肪累积增加，脂滴增多，细胞肥大，并且血清谷丙转氨酶（ALT）水平增加（P < 0.0001）；经双歧杆菌治疗后，NAFLD小鼠肝脏脂肪积累减少并且血清ALT水平降低（P < 0.0001）。肠道微生物群分析在3个组别中鉴定出共有菌种348种。微生物功能分析显示，双歧杆菌组肠道微生物代谢功能水平介于对照组和NAFLD组之间；双歧杆菌治疗能改变NAFLD模型中特定肠道微生物的丰度变化，并且这些肠道微生物可能参与了谷胱甘肽代谢、嘌呤代谢和鞘脂代谢等途径调节机体的代谢功能。 结论 双歧杆菌三联活菌在小鼠模型中可能通过调节肠道微生物组的功能改善NAFLD，并且双歧杆菌三联活菌有改善机体脂代谢紊乱的潜力。 -
关键词:
- 非酒精性脂肪性肝（NAFLD） /
- 宏基因组 /
- 双歧杆菌三联活菌 /
- 高脂饮食
Abstract:Objective To investigate the therapeutic effects of probiotics containing Bifidobacterium on a mouse model of non-alcoholic fatty liver disease （NAFLD） and its impact on the gut microbiota. Methods C57BL/6J mice were divided into three experimental groups. The control group （n = 10） of mice was fed a normal diet, the NAFLD model group （n = 10） was fed a high-fat diet, and the probiotics group （n = 10） was given Bifidobacteriumtrisporus treatment while using NAFLD model mice. HE staining was performed on paraffin sections of mouse liver from different groups at the end of the experiment to observe pathological changes. The 16S rRNA gene sequencing technique was used to detect the gut microbiota in each group of mice, followed by bioinformatics analysis. Results Compared with the control group, NAFLD mice showed increased accumulation of liver fat, increased number of lipid droplets, enlarged cells, and increased levels of serum alanine aminotransferase （ALT）（P < 0.0001）. After treatment with Bifidobacteriumtrisporus, the accumulation of liver fat in NAFLD mice decreased and serum ALT levels decreased （P < 0.0001）. Microbiota analysis of the intestinal tract identified 348 common species of microorganisms in the three groups. Functional analysis of the microbiota showed that the metabolic function of the intestinal microbiota in the Bifidobacterium group was intermediate between the control group and the NAFLD group; Bifidobacterium treatment was able to alter the abundance of specific intestinal microorganisms in the NAFLD model, and these intestinal microorganisms may be involved in the regulation of metabolic function pathways such as glutathione metabolism, purine metabolism, and sphingolipid metabolism in the body. Conclusion Bifidobacterium may improve NAFLD by regulating the function of the intestinal microbiota in a mouse model, and it has the potential to improve disordered lipid metabolism in the body. -
生物导向型预备技术（biologically oriented preparation technique，BOPT）由Loi在2008年首次提出[1]，通过临时修复体对牙龈塑形，获得满意的牙龈形态后再在永久修复体上复制穿龈部分的准确形态。BOPT理念倡导牙体预备中并不形成明确的肩台，根据牙周参数在颈部区域勾勒牙冠，以利于更加自由的调整牙龈形态。与其他牙体预备技术相比，BOPT 技术在准备印模、临时牙冠重衬和创建牙冠轮廓直至最终修复的过程更简单、更快捷[1]。
BOPT是传统水平精加工线的可靠替代方案。使用BOPT后，牙龈组织可根据修复体出现的形状进行建模，形成牙周保护组织，特别是形成新的牙周组织，在结构上等同于正常的牙周组织，以确保没有炎症的健康环境[2]。
现在BOPT已广泛应用于口腔各类修复方案中[3]，本文将从贴面、全冠、种植及总结4个方面综合论述BOPT的应用现状。
1. BOPT在贴面中的应用
BOPT技术代表了目前口腔修复领域牙体预备的新趋势。这种技术与陶瓷贴面一起使用，可将牙龈龈缘线的稳定性转移到陶瓷贴面上，从而稳定牙龈组织并避免不必要的牙龈退缩[4]。
研究发现[5]，进行正确的牙周分析后，在没有终点线的情况下进行牙体预备可纠正牙龈不对称，这有助于成功维持软组织稳定。作为常规治疗的替代方案，使用BOPT在前牙区应用陶瓷贴面有助于牙周组织的改建和改善牙龈对称性。在贴面中使用 BOPT，牙体制备不设置水平终点线，软组织能在正确的位置再生，进而达到良好的适应和稳定，甚至可以纠正软组织异常和不对称。与传统的制备技术相比，BOPT在牙体预备过程中产生更大程度的牙龈增厚，无论患者呈现薄牙龈生物型还是厚牙龈生物型，都会降低由于血管形成增加而导致牙龈移位的风险。
2. BOPT在全冠中的应用
有研究表明，固定义齿的留存率与冠修复材料无密切相关性[6]。固定义齿的牙龈边缘位置一直是口腔修复领域争论的热点[7-8]。牙龈修复位置不当会导致牙周病及龋病的发生[9]。 Maniatopoulos等[10]的前瞻性研究表明，BOPT形成良好的修复体周围结构，提高了牙龈边缘的稳定性，增加修复体周围的牙龈厚度。
固定义齿修复最常见的并发症之一是牙龈边缘的顶端迁移，这可能与牙龈边缘位置和牙体预备类型等因素有关。Serra-Pastor等[11]对25名患者使用BOPT制备前牙后行固定桥修复，在进行长达6 a的追踪评估后得出结论：使用BOPT制备的牙齿后行固定桥修复，基牙具有良好的牙周健康和牙龈边缘稳定性，且牙龈无退缩，固定桥留存率为100%。
有学者[12]使用BOPT进行单个氧化锆冠修复前牙的牙周组织评估和患者满意度进行为期 6 a的前瞻性临床研究，研究表明在前牙区用BOPT制备的牙齿在修复体周围特别在牙龈边缘的稳定性和牙龈厚度增加方面，表现出良好的牙周形态。用BOPT制备的单冠获得了优异的临床留存率，并且在6 a后获得较高的患者满意度。
3. BOPT在种植修复中的应用
种植体的长期成功率部分取决于修复体和种植体颈部周围建立的软组织圈，这一结构可以提供有效的密封，防止细菌入侵后导致的种植体周围炎[13]。临床医生能够决定和调整种植体周围软组织的边缘水平，从而改变修复体冠颈缘，提供软组织的适应性形态，软组织以受控方式侵入龈沟，从而确保种植体周围组织的健康，提供种植体周围硬组织和软组织的稳定性[14-16]。
良好的种植体周围软组织可防止牙龈组织萎缩，并可作为防止食物嵌塞的机械屏障，这是植入物支撑修复体不可避免的常见问题[17]。BOPT通过胶原纤维增强上皮组织附着和种植体周围黏膜密封，胶原纤维随着时间的推移保持牙龈组织的稳定和厚实，更加有效保护种植体周围骨组织[18-20]。
在种植体上粘结修复体与固定修复体粘结到天然牙齿的设计和美学原则一致[21]。BOPT同样适用于粘结在种植体上的固定修复体[22-23]，确保种植体周围组织健康，预防牙周组织发生炎症[21]。Cabanes-Gumbau 等[24]的研究表明，使用BOPT行种植修复，10月后种植体周围软组织厚度显着增加[25]。种植体周围软组织的形态功能及骨质流失与使用的修复体类型有关，与传统的种植体相比，采用BOPT的种植体修复表现出更好的种植体周围软组织形态，种植体周围骨质流失也较少[26-27]。
4. 小结
综上所述，BOPT在贴面、全冠及种植修复后修复体周围都表现出良好的牙周组织形态，大大提高修复的成功率及患者满意度。使用BOPT预备的修复体其周围软组织表现出良好的形态，牙龈增厚，边缘组织稳定性增加，并发症的发生率较低。在需要更换旧修复体的情况下，BOPT 是一种很好的治疗选择，表现出良好的牙周组织形态、牙龈增厚和边缘稳定性增加[8]。许多研究已经证明使用该技术治疗的牙齿的牙周组织状况有所改善[2]。随着BOPT研究的深入，BOPT的应用范围越来越广阔[3]。BOPT还可与全数字工作流程相结合，矫正和重塑牙龈形态[28]。
尽管BOPT有一定的局限性，对于牙龈厚度不足难以确保生物宽度有足够的空间行种植修复的临床病例，BOPT并不适用。此外，该技术非常耗时，因为在第2阶段手术期间，临床医生必须调整临时基台，这需要丰富的临床经验，对新手医生是极大的挑战[29]。但BOPT在临床中应用广泛，促进牙周组织再生，恢复良好边缘形态，具有良好的应用前景，值得临床推广。
-
图 1 双歧杆菌改善小鼠NAFLD发生
A：对照组，NAFLD组和双歧杆菌组小鼠肝脏HE染色（200×）；B：对照组， NAFLD组和双歧杆菌组小鼠血清中谷丙转氨酶（ALT）活性检测。对照组与 NAFLD组ALT比较，****P < 0.0001；NAFLD组和双歧杆菌组ALT比较，****P < 0.0001。
Figure 1. Bifidobacterium improves the NAFLD in the mouse model
图 2 各组小鼠粪便样本鉴定微生物种类韦恩图
A：对照组；B：NAFLD组；C：双歧杆菌组。
Figure 2. Venn diagram of microbial species identified from fecal samples of mice in each group
图 3 各组小鼠粪便样本鉴定微生物种类丰度分布
A：对照组；B：NAFLD组；C：双歧杆菌组。
Figure 3. Abundance distribution of identified microbial species in fecal samples in each group
图 4 各组小鼠粪便样本宏基因组数据测序基因片段功能富集分析
A：对照组；B：NAFLD组；C：双歧杆菌组。
Figure 4. Functional enrichment analysis of gene fragments in the metagenomic data of fecal samples from mice in each group
-
[1] Wagner R,Eckstein S S,Yamazaki H,et al. Metabolic implications of pancreatic fat accumulation[J]. Nat Rev Endocrinol,2022,18(1):43-54. doi: 10.1038/s41574-021-00573-3 [2] Kwan S Y,Jiao J,Joon A,et al. Gut microbiome features associated with liver fibrosis in Hispanics,a population at high risk for fatty liver disease[J]. Hepatology,2022,5(4):955-967. [3] Kolodziejczyk A A,Zheng D,Shibolet O,et al. The role of the microbiome in NAFLD and NASH[J]. EMBO Mol Med,2019,11(2):e9302. doi: 10.15252/emmm.201809302 [4] Safari Z,Gérard P. The links between the gut microbiome and non-alcoholic fatty liver disease (NAFLD)[J]. Cell Mol Life Sci,2019,76(8):1541-1558. doi: 10.1007/s00018-019-03011-w [5] Hu H,Lin A,Kong M,et al. Intestinal microbiome and NAFLD: Molecular insights and therapeutic perspectives[J]. J Gastroenterol,2020,55(2):142-158. doi: 10.1007/s00535-019-01649-8 [6] Aron-Wisnewsky J,Vigliotti C,Witjes J,et al. Gut microbiota and human NAFLD: Disentangling microbial signatures from metabolic disorders[J]. Nat Rev Gastroenterol Hepatol,2020,17(5):279-297. doi: 10.1038/s41575-020-0269-9 [7] Lau L H S,Wong S H. Microbiota,obesity and NAFLD[J]. Adv Exp Med Biol,2018,1061:111-125. [8] Canfora E E,Meex R C R,Venema K,et al. Gut microbial metabolites in obesity,NAFLD and T2DM[J]. Nat Rev Endocrinol,2019,15(5):261-273. doi: 10.1038/s41574-019-0156-z [9] Fan Y,Pedersen O. Gut microbiota in human metabolic health and disease[J]. Nat Rev Microbiol,2021,19(1):55-71. doi: 10.1038/s41579-020-0433-9 [10] Bana B,Cabreiro F. The microbiome and aging[J]. Annu Rev Genet,2019,53:239-261. doi: 10.1146/annurev-genet-112618-043650 [11] Suez J,Zmora N,Segal E,et al. The pros,cons,and many unknowns of probiotics[J]. Nat Med,2019,25(5):716-729. doi: 10.1038/s41591-019-0439-x [12] Qian X,Si Q,Lin G,et al. Bifidobacterium adolescentis is effective in relieving type 2 diabetes and may be related to its dominant core genome and gut microbiota modulation capacity[J]. Nutrients,2022,14(12):2479. doi: 10.3390/nu14122479 [13] Fang Z,Pan T,Li L,et al. Bifidobacterium longum mediated tryptophan metabolism to improve atopic dermatitis via the gut-skin axis[J]. Gut Microbes,2022,14(1):2044723. doi: 10.1080/19490976.2022.2044723 [14] Eng J M,Estall J L. Diet-induced models of non-alcoholic fatty liver disease: Food for thought on sugar,fat,and cholesterol[J]. Cells,2021,10(7):1805.P. doi: 10.3390/cells10071805 [15] Zeng H,Larson K J,Cheng WH,et al. Advanced liver steatosis accompanies an increase in hepatic inflammation,colonic,secondary bile acids and Lactobacillaceae/Lachnospiraceae bacteria in C57BL/6 mice fed a high-fat diet[J]. J Nutr Biochem,2020,78:108336. doi: 10.1016/j.jnutbio.2019.108336 [16] Yuan G,Tan M,Chen X. Punicic acid ameliorates obesity and liver steatosis by regulating gut microbiota composition in mice[J]. Food Funct,2021,12(17):7897-7908. doi: 10.1039/D1FO01152A [17] Wang H,Wang Q,Yang C,et al. Bacteroides acidifaciens in the gut plays a protective role against CD95-mediated liver injury[J]. Gut Microbes,2022,14(1):2027853. doi: 10.1080/19490976.2022.2027853 [18] Nishimura N,Kaji K,Kitagawa K,et al. Intestinal permeability is a mechanical rheostat in the pathogenesis of liver cirrhosis[J]. Int J Mol Sci,2021,22(13):6921. doi: 10.3390/ijms22136921 [19] Do M H,Oh M J,Lee H B,et al. Bifidobacterium animalis ssp. lactis MG741 reduces body weight and ameliorates nonalcoholic fatty liver disease via improving the gut permeability and amelioration of inflammatory cytokines[J]. Nutrients,2022,14(9):1965. doi: 10.3390/nu14091965 [20] Wang W,Xu A L,Li Z C,et al. Combination of probiotics and salvia miltiorrhiza polysaccharide alleviates hepatic steatosis via gut microbiota modulation and insulin resistance improvement in high fat-induced NAFLD mice[J]. Diabetes Metab J,2020,44(2):336-348. doi: 10.4093/dmj.2019.0042 [21] Yang X,Mo W,Zheng C,et al. Alleviating effects of noni fruit polysaccharide on hepatic oxidative stress and inflammation in rats under a high-fat diet and its possible mechanisms[J]. Food Funct,2020,11(4):2953-2968. doi: 10.1039/D0FO00178C [22] Wang X,Shi L,Wang X,et al. MDG-1,an Ophiopogon polysaccharide,restrains process of non-alcoholic fatty liver disease via modulating the gut-liver axis[J]. Int J Biol Macromol,2019,141(3):1013-1021. [23] Tong A J,Hu R K,Wu L X,et al. Ganoderma polysaccharide and chitosan synergistically ameliorate lipid metabolic disorders and modulate gut microbiota composition in high fat diet-fed golden hamsters[J]. J Food Biochem,2020,44(1):e13109. [24] Dehhaghi M, Kazemi Shariat Panahi H, Guillemin GJ. Microorganisms, tryptophan metabolism, and kynurenine pathway: A complex interconnected loop influencing human health status [J]. Int J Tryptophan Res, 2019, 12（2）: 1178646919852996. [25] Arnoriaga-Rodríguez M,Mayneris-Perxachs J,Burokas A,et al. Obesity impairs short-term and working memory through gut microbial metabolism of aromatic amino acids[J]. Cell Metab,2020,32(4):548-560,e7. doi: 10.1016/j.cmet.2020.09.002 [26] Zhao Z H,Xin F Z,Xue Y,et al. Indole-3-propionic acid inhibits gut dysbiosis and endotoxin leakage to attenuate steatohepatitis in rats[J]. Exp Mol Med,2019,51(9):1-14. -
下载:
点击查看大图
计量
- 文章访问数: 1771
- HTML全文浏览量: 1065
- PDF下载量: 31
- 被引次数: 0
