老年人大便性状与肠道菌群的相关性分析

doi:10.3877/cma.j.issn.1674-6880.2018.04.001

目的

比较老年人不同大便性状与肠道菌群间的相关性。

方法

将23例老年患者根据bristol大便分型分成便秘组（13例）及便溏组（10例），并同期选择9例健康人群为对照组。留取新鲜粪便样本并提取DNA，对细菌16S rDNA基因V4区进行粪便微生物DNA测序，对测序结果进行生物信息学分析。同时，采用酶联免疫吸附试验（ELISA）检测肿瘤坏死因子α（TNF-α）、白细胞介素10（IL-10）、IL-17、IL-22、白三烯B4（LTB-4）及前列腺素E2（PGE2）水平。采用Spearman法分析肠道菌群相对丰度与大便性状的相关性。

结果

三组患者间Shannon指数和Simpson指数比较，差异均有统计学意义（H= 8.449，P= 0.015；H= 6.884，P= 0.032），且便秘组患者Shannon指数[6.4（5.8，6.6）vs. 5.0（4.8，5.6）]和Simpson指数[0.95（0.93，0.97）vs. 0.88（0.83，0.94）]均显著高于便溏组患者（P均< 0.05）。但三组患者间Ace指数和Chao指数比较，差异均无统计学意义（H= 3.821，P= 0.148；H= 3.516，P= 0.172）。三组患者间柯林斯均属（H= 6.599，P= 0.037）、丁酸弧菌属（H = 8.707，P = 0.013）、嗜胆菌属（H = 7.354，P = 0.025）、厌氧棍状均属（H = 6.179，P = 0.046）、巨球形均属（H= 6.095，P= 0.047）的相对丰度比较，差异均有统计学意义。而三组患者间TNF-α、IL-10、IL-17、IL-22、LTB-4及PGE2的比较，差异均无统计学意义（F = 0.442、1.034、3.217、0.957、0.595、1.440，P均> 0.05）。同时，柯林斯菌属、丁酸弧菌属、嗜胆菌属、厌氧棍状菌属与大便稠度显著正相关（r= 0.455、0.415、0.449、0.411，P= 0.009、0.018、0.010、0.019）。

结论

在老年人中，大便性状的不同与肠道菌群的变化有关。

关键词: 粪便, 菌群, 肠道, 老年人

Objective

To investigate the relationship between fecal characteristics and intestinal microflora in the elderly.

Methods

Totally 23 elderly patients were divided into the constipation group (13 cases) and loose stool group (10 cases) according to bristol stool classification, and 9 healthy people were enrolled as the control group. Fresh fecal samples were taken and DNA was extracted. The bacterial microbial DNA was sequenced in the V4 region of 16S rDNA gene, and the sequencing results were analyzed by bioinformatics. The enzyme-linked immunosorbant assay (ELISA) was adopted to detect levels of tumor necrosis factor-alpha (TNF-α), interleukin 10 (IL-10), IL-17, IL-22, leukotriene B4 (LTB-4), and prostaglandin E2 (PGE2). The Spearman method was used to analyze the relationship between fecal characteristics and in testinal microflora.

Results

Shannon and Simpson indices were significantly different among three groups (H= 8.449, P= 0.015; H= 6.884, P= 0.032), and the Shannon index [6.4 (5.8, 6.6) vs. 5.0 (4.8, 5.6)] and Simpson index [0.95 (0.93, 0.97) vs. 0.88 (0.83, 0.94)] were significantly higher in the constipation group than in loose stool group (both P< 0.05). However, there were no significant differences in Ace and Chao indices among three groups (H= 3.821, P= 0.148; H= 3.516, P= 0.172). The relative abundance of Collinsella (H= 6.599, P= 0.037), Anaerostipes (H= 8.707, P= 0.013), Bilophila (H= 7.354, P= 0.025), Anaerotruncus (H= 6.179, P= 0.046) and Megasphaera (H= 6.095, P= 0.047) was significantly different among three groups. The levels of TNF-α, IL-10, IL-17, IL-22, LTB-4 and PGE2 showed no significant differences among them (F= 0.442, 1.034, 3.217, 0.957, 0.595, 1.440; all P> 0.05). Meanwhile, the abundance of Collinsella, Anaerostipes, Bilophila and Anaerotruncus was positively correlated with stool consistency (r= 0.455, 0.415, 0.449, 0.411; P= 0.009, 0.018, 0.010, 0.019) .

Conclusion

Fecal characteristics is associated with intestinal microflora in the elderly.

Key words: Feces, Microflora, Intestinal tract, Aged

表1 三组老年患者间菌群丰富度和多样性的比较[M（P₂₅，P₇₅）]

组别	例数	Ace指数	Chao指数	Shannon指数	Simpson指数
便溏组	10	1 653(1 432,1 882)	1 670(1 540,1 888)	5.0(4.8,5.6)	0.88(0.83,0.94)
对照组	9	1 864(1 482,1 956)	1 772(1 512,1 900)	6.1(5.2,6.3)	0.95(0.92,0.96)
便秘组	13	1 936(1 841,2 209)	1 867(1 808,2 153)	6.4(5.8,6.6)^a	0.95(0.93,0.97)^a
H值	?	3.821	3.516	8.449	6.884
P值	?	0.148	0.172	0.015	0.032

表1 三组老年患者间菌群丰富度和多样性的比较[M（P₂₅，P₇₅）]

组别	例数	Ace指数	Chao指数	Shannon指数	Simpson指数
便溏组	10	1 653(1 432,1 882)	1 670(1 540,1 888)	5.0(4.8,5.6)	0.88(0.83,0.94)
对照组	9	1 864(1 482,1 956)	1 772(1 512,1 900)	6.1(5.2,6.3)	0.95(0.92,0.96)
便秘组	13	1 936(1 841,2 209)	1 867(1 808,2 153)	6.4(5.8,6.6)^a	0.95(0.93,0.97)^a
H值	?	3.821	3.516	8.449	6.884
P值	?	0.148	0.172	0.015	0.032

表2 三组老年患者菌群相对丰度的比较[M（P₂₅，P₇₅）]

组别	例数	柯林斯均属(%)	丁酸弧菌属(%)	嗜胆菌属(%)	厌氧棍状菌属（‰）	巨球形菌属(%)
便溏组	10	0.12(0.11,0.19)	0.016(0.093,0.035)	0.06(0.03,0.24)	0.029(0.019,0.081)	0.123(0.020,0.386)
对照组	9	0.17(0.06,0.90)	0.074(0.029,0.160)^a	0.19(0.17,0.34)	0.144(0.057,0.164)^a	0.350(0.096,0.762)
便秘组	13	0.28(0.21,0.38)^a	0.077(0.045,0.119)^a	0.30(0.14,0.45)^a	0.077(0.059,0.199)	0.048(0.020,0.170)^b
H值	?	6.599	8.707	7.354	6.179	6.095
P值	?	0.037	0.013	0.025	0.046	0.047

表2 三组老年患者菌群相对丰度的比较[M（P₂₅，P₇₅）]

组别	例数	柯林斯均属(%)	丁酸弧菌属(%)	嗜胆菌属(%)	厌氧棍状菌属（‰）	巨球形菌属(%)
便溏组	10	0.12(0.11,0.19)	0.016(0.093,0.035)	0.06(0.03,0.24)	0.029(0.019,0.081)	0.123(0.020,0.386)
对照组	9	0.17(0.06,0.90)	0.074(0.029,0.160)^a	0.19(0.17,0.34)	0.144(0.057,0.164)^a	0.350(0.096,0.762)
便秘组	13	0.28(0.21,0.38)^a	0.077(0.045,0.119)^a	0.30(0.14,0.45)^a	0.077(0.059,0.199)	0.048(0.020,0.170)^b
H值	?	6.599	8.707	7.354	6.179	6.095
P值	?	0.037	0.013	0.025	0.046	0.047

表3 三组老年患者间炎症因子表达水平的比较（±s）

组别	例数	TNF-α(μg/L)	IL-10(ng/L)	IL-17(ng/L)	IL-22(ng/L)	LTB-4(ng/L)	PGE2(ng/L)
便溏组	10	979 ± 919	15.8 ± 1.5	10.3 ± 1.5	47 ± 33	13.1 ± 2.2	42 ± 29
对照组	9	1 998 ± 3 516	16.4 ± 2.2	13.4 ± 1.9	42 ± 22	14.2 ± 2.1	31 ± 5
便秘组	13	1 496 ± 2 102	14.0 ± 3.9	13.0 ± 3.9	65 ± 54	13.0 ± 3.1	29 ± 6
H值	?	0.442	1.034	3.217	0.957	0.595	1.440
P值	?	0.647	0.378	0.057	0.396	0.559	0.256

表3 三组老年患者间炎症因子表达水平的比较（±s）

组别	例数	TNF-α(μg/L)	IL-10(ng/L)	IL-17(ng/L)	IL-22(ng/L)	LTB-4(ng/L)	PGE2(ng/L)
便溏组	10	979 ± 919	15.8 ± 1.5	10.3 ± 1.5	47 ± 33	13.1 ± 2.2	42 ± 29
对照组	9	1 998 ± 3 516	16.4 ± 2.2	13.4 ± 1.9	42 ± 22	14.2 ± 2.1	31 ± 5
便秘组	13	1 496 ± 2 102	14.0 ± 3.9	13.0 ± 3.9	65 ± 54	13.0 ± 3.1	29 ± 6
H值	?	0.442	1.034	3.217	0.957	0.595	1.440
P值	?	0.647	0.378	0.057	0.396	0.559	0.256

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Correlation between fecal characteristics and intestinal microflora in the elderly

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Correlation between fecal characteristics and intestinal microflora in the elderly