姜黄素干预小鼠肠道菌群并抑制结直肠癌变的实验研究

doi:10.3877/cma.j.issn.2095-3224.2018.06.007

目的

拟通过监测姜黄素干预小鼠肠癌发生发展过程中肠道菌群的变化，探究姜黄素对小鼠肠道菌群的影响，以及可能的抗癌作用机制及作用靶点。

方法

将25只6周龄C57BL/6小鼠随机分为基础饮食组（BD）5只，AOM/DSS造模组（MO）10只，姜黄素干预造模组（CU）10只，分别于实验前和处死前收集小鼠粪便进行DNA提取及高通量菌群测序。

结果

T1点时（干预前）各组小鼠肠道菌群的多样性指数差异无统计学意义，T2点时（干预后）MO组的多样性指数更高，高于其他2组，与BD组差异有统计学意义（t=2.73，P=0.02），而与CU组比较差异无统计学意义。T2点时各组的多样性指数均较T1点时明显升高，其中CU组与BD在T2点时多样性指数更接近，均小于造模组。造模前后菌群在属水平发生了较大变化，CU组较MO组肠道菌群变化要小，而且拟杆菌类和疣微菌门较MO组变化较大。MO小鼠体重增量平均要少于CU组，CU组小鼠结直肠肿瘤的成瘤数量、瘤体体积，明显少于MO组。

结论

在小鼠结直肠癌发生过程中，姜黄素能够维持小鼠肠道菌群的稳定性，降低了小鼠肠道肿瘤发生率，可能为抑制结直肠癌发生发展的机制之一。

关键词: 结直肠肿瘤, 姜黄素, 肠道菌群, 测序

Objective

To explore the effect of curcumin on intestinal flora in mice, as well as anti-cancer mechanisms and targets by monitoring the changes of intestinal flora during the development of colorectal cancer in mice.

Methods

Twenty-five 6-week-old C57BL/6 mice were randomly divided into basic diet group (BD) with 5, AOM/DSS model group (MO) with 10, and curcumin intervention model group (CU) with 10. DNA extraction and high-throughput microbiota sequencing were carried out before the experiment and before the execution.

Results

There was no significant difference in the diversity index of mice in each group at point T1 (before intervention), and the diversity index of MO group at point T2 (after intervention) was higher than that of the other 2 groups, with a significant difference from BD group (t=2.73, P=0.02) and no significant difference from CU group. The diversity index of each group at T2 was significantly higher than that at T1, in which the diversity index of CU group and BD group were more similar at T2, and were all smaller than the MO group. Before and after the molds were made, the flora of the genus changed greatly, and the intestinal flora of the CU group changed less than that of the MO group, especially the Bacteroidetes and Verrucomicrobia. The average weight increment of MO mice was lower than that of CU group, and the number of tumor formation and tumor volume of colorectal tumors in CU group were significantly lower than that of MO group.

Conclusion

Curcumin can maintain the stability of intestinal flora and reduce the incidence of intestinal tumor in mice during the occurrence of colorectal cancer, which indicating that it is possible one of the mechanisms to inhibit the occurrence and development of colorectal cancer.

Key words: Colorectal neoplasms, Curcumin, Gut microbiota, Sequencing

图1 小鼠体重增长图

图2 造模组及姜黄素组成瘤图片

图3 成瘤数目及载瘤负荷图

图4 序列长度分布图

图5 稀释曲线及丰度分布曲线图

表1 T1点（干预前）Alpha多样性指数表

组别	例数	?	辛普森指数	香农指数
BD组	5	?	0.17±0.12	2.43±0.81
MO组	8	?	0.37±0.15	1.52±0.75
CU组	8	?	0.33±0.16	1.88±0.86
整体比较	F，P	?	6.73，0.025	4.75，0.031
各组比较	LSD-t，P	BD vs. MO	2.51，0.05	2.07，0.06
?	?	BD vs. CU	1.91，0.09	1.15，0.26
?	?	MO vs. CU	0.51，0.33	0.89，0.38

表1 T1点（干预前）Alpha多样性指数表

组别	例数	?	辛普森指数	香农指数
BD组	5	?	0.17±0.12	2.43±0.81
MO组	8	?	0.37±0.15	1.52±0.75
CU组	8	?	0.33±0.16	1.88±0.86
整体比较	F，P	?	6.73，0.025	4.75，0.031
各组比较	LSD-t，P	BD vs. MO	2.51，0.05	2.07，0.06
?	?	BD vs. CU	1.91，0.09	1.15，0.26
?	?	MO vs. CU	0.51，0.33	0.89，0.38

表2 T2点（干预后）Alpha多样性指数表

组别	例数	?	辛普森指数	香农指数
BD组	5	?	0.09±0.03	3.16±0.31
MO组	8	?	0.06±0.02	3.48±0.30
CU组	8	?	0.10±0.06	3.16±0.42
整体比较	F，P	?	9.93，0.001	6.84，0.042
各组比较	LSD-t，P	BD vs. MO	2.73，0.02	1.84，0.08
?	?	BD vs. CU	1.79，0.83	0.00，0.98
?	?	MO vs. CU	0.34，0.12	1.75，0.09

表2 T2点（干预后）Alpha多样性指数表

组别	例数	?	辛普森指数	香农指数
BD组	5	?	0.09±0.03	3.16±0.31
MO组	8	?	0.06±0.02	3.48±0.30
CU组	8	?	0.10±0.06	3.16±0.42
整体比较	F，P	?	9.93，0.001	6.84，0.042
各组比较	LSD-t，P	BD vs. MO	2.73，0.02	1.84，0.08
?	?	BD vs. CU	1.79，0.83	0.00，0.98
?	?	MO vs. CU	0.34，0.12	1.75，0.09

图6 门水平小鼠肠道菌群柱状图

图7 各组门水平小鼠肠道菌群差异图。7A：BD组干预前后门水平菌群差异；7B：CU组干预前后门水平菌群差异；7C：MO组干预前后门水平菌群差异；7D：CU组与MO组干预后门水平菌群差异

图8 各组属水平小鼠肠道菌群差异图。8A：MO组干预前后属水平菌群差异；8B：CU组干预前后属水平菌群差异；8C：CU组与MO组干预后属水平菌群差异

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Structural shift of gut microbiota during chemo-preventive effects of curcumin on colorectal carcinogenesis in mice

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Structural shift of gut microbiota during chemo-preventive effects of curcumin on colorectal carcinogenesis in mice