Abstract: Objective To observe the effect of d-limonene on intestinal microbes in nude mice bearing MCF-7 breast cancer. Method After 30 nude mice were inoculated with MCF-7, dextro-limonene was given at 80 mg / kg, 160 mg / kg, and 320 mg / kg for 6 weeks. Fecal genomic DNA was extracted, and PCR amplification was performed to construct after the library was quantified by Qubit and Q-PCR, the NovaSeq6000 was used for online sequencing, and each sample data was separated from the offline data. After strict filtering, high-quality Clean Tags were obtained, and the chimera sequence was removed to obtain the final after the effective data ( effective tags) , analyze the microbial diversity. Result The bacterial species and abundance decreased after tumor inoculation, and the intestinal flora species and abundance increased after d-limonene treatment. The tumor group was close to the phylum Firmicutes and Bacteroides at the phylum level, and the bisphenol group A and the right number of Bacteroides in the limonene group increased. At the genus level, compared with the tumor control group ( group H ) , Lactobacillus in the bisphenol A and dextro-limonene group decreased significantly, and the isovalerobacter genus increased significantly. Parabacteroides diundi was in the tumor group and bisphenol group A dropped significantly, but the bacteria increased after feeding d-limonene. Conclusion D-limonene significantly affects the number and abundance of intestinal flora in nude mice with breast cancer. Bacteroides and Isovaleri are the dominant flora.

Key words: d-limonene, MCF-7 cells, intestinal microorganisms, anti-tumor

摘要： 目的 观察右旋柠檬烯对荷 MCF-7 乳腺癌的裸鼠肠道微生物的影响。 方法 30 只裸鼠接种 MCF-7 后,随机分为 5 组,模型组(每天给予蒸馏水) ,双酚 A+右旋柠檬烯低、中、高剂量组( 80、160、320 mg / kg) ,双酚 A 组( 1 mL /kg) ,每组 6 只;每天灌胃给药 1 次,连续 6 周,接取新鲜粪便,提取粪便基因组 DNA,并进行 PCR 扩增,构建文库后上机测序,下机数据中拆分出各样本数据,经过严格的过滤处理得到高质量的 Clean Tags,删除嵌合体序列后得到有效数据( effective tags)用于微生物多样性的分析。 结果 接种肿瘤后细菌种类和丰度下降,右旋柠檬烯治疗后肠道菌群种类和丰度增加,肿瘤组在门水平上厚壁菌门与拟杆菌门比例接近,双酚 A 组和右旋柠檬烯组的拟杆菌门菌数量增加。 在属水平上,与肿瘤对照组( H 组)相比,双酚 A 及右旋柠檬烯组的乳杆菌显著下降,异戊酸杆菌属显著增加,狄氏副拟杆菌在肿瘤组和双酚 A 组显著下降,而喂食右旋柠檬烯后该菌增加。 结论 右旋柠檬烯显著影响了乳腺癌裸鼠肠道菌群的数量和丰度,拟杆菌门和异戊酸杆菌是优势菌群。

关键词: 右旋柠檬烯, MCF-7 细胞, 肠道微生物, 抗肿瘤