基于流式细胞术研究叶绿素对肠道细菌增殖的影响

郑红莉; 王潇; 杜梦璇; 刘畅; 张燕

doi:10.13386/j.issn1002-0306.2021120296

基于流式细胞术研究叶绿素对肠道细菌增殖的影响

Effect of Chlorophyll on the Proliferation of Intestinal Bacteria Based on Flow Cytometry Method

摘要

摘要: 为在菌种水平上探究叶绿素对特定肠道细菌增殖的影响，本研究首先探究了流式细胞术（Flow cytometry，FCM）对肠道细菌计数的可行性并与稀释涂布平板法进行比较，随后利用FCM探究了叶绿素对珀氏解黄酮菌（Flavonifractor plautii）、普通拟杆菌（Bacteroides vulgatus）、假长双岐杆菌（Bifidobacterium pseudolongum）和鼠乳杆菌（Lactobacillus murinus）增殖的影响。结果表明，FCM可实现对活细菌和死细菌的同时检测，FCM活菌计数结果与稀释涂布平板法测定的活菌数之间具有很好的相关性（r>0.8，P<0.001），但FCM检测效率更高、检测结果更接近于真实值。100 µg/mL的叶绿素可高度显著促进Flavonifractor plautii的增殖（P<0.01），而300和500 µg/mL的叶绿素可极显著抑制Flavonifractor plautii的增殖（P<0.001）；100 µg/mL的叶绿素可极显著促进Bacteroides vulgatus的增殖（P<0.001），而300和500 µg/mL的叶绿素可极显著抑制Bacteroides vulgatus的增殖（P<0.001）；叶绿素可极显著抑制Bifidobacterium pseudolongum的增殖（P<0.001），且浓度越高抑制作用越强；叶绿素可极显著促进Lactobacillus murinus的增殖（P<0.001），且浓度越高促进作用越强。本研究为阐明膳食叶绿素对肠道菌群生长的影响规律提供了重要的理论数据。

Abstract: In order to explore the effect of chlorophyll on the proliferation of specific intestinal bacteria at the bacterial species level, this study firstly explored the feasibility of flow cytometry (FCM) to count intestinal bacteria and compared it with the dilution plating method. Then, the effects of chlorophyll on the proliferation of Flavonifractor plautii, Bacteroides vulgatus, Bifidobacterium pseudolongum and Lactobacillus murinus were investigated by FCM. The results showed that FCM could detect live bacteria and dead bacteria at the same time. There was a good correlation between the count of live bacteria by FCM and the number of live bacteria determined by the dilution coating method (r>0.8, P<0.001). However, detection efficiency of FCM was higher, and the detection results were closer to the real value. 100 µg/mL chlorophyll highly significantly promoted the proliferation of Flavonifractor plautii (P<0.01), while 300 and 500 µg/mL chlorophyll extremely significantly inhibited the proliferation of Flavonifractor plautii (P<0.001). 100 µg/mL chlorophyll extremely significantly promoted the proliferation of Bacteroides vulgatus (P<0.001), while 300 and 500 µg/mL chlorophyll extremely significantly inhibited the proliferation of Bacteroides vulgatus (P<0.001). Chlorophyll could extremely significantly inhibit the proliferation of Bifidobacterium pseudolongum (P<0.001), and the higher the concentration was, the stronger the inhibitory effect was. Chlorophyll could extremely significantly promote the proliferation of Lactobacillus murinus (P<0.001), and the higher the concentration was, the stronger the promoting effect was. This study would provide important theoretical data for elucidating the effect of dietary chlorophyll on the growth of gut microbiota.