高脂饲料诱导的肥胖大鼠肠道内硫酸盐还原菌的定量检测

Quantification of sulfate-reducing bacteria in the gut of high-fat diet-induced obese rats using real-time qPCR

目的检测高脂饲料诱导大鼠肥胖过程中,大鼠肠道内硫酸盐还原菌(sulfate-reducing bacteria,SRB)的数量变化,为研究SRB与肥胖的关系提供参考。方法 20只Wistar大鼠随机分为2组(每组10只),一组饲喂高脂饲料(HFD组)18周,另一组饲喂正常饲料(NCD组,即对照组)18周。以编码腺苷酰硫酸还原酶α亚基的基因(aprA)作为分子标记,通过荧光定量PCR的方法检测两组大鼠在0、8和18周,肠道内SRB的数量变化;同时,以16S rRNA基因作为标记基因定量大鼠肠道内总菌的数量,以计算肠道内SRB在总菌中的比例变化。结果分组饲喂8周后,高脂饲料饲喂组大鼠的体重与正常饲料组相比显著升高。对SRB的定量结果显示,饲喂8周和18周,高脂饲料组大鼠肠道内SRB的数量和含量与正常饲料组相比显著升高。结论大鼠肠道中的硫酸盐还原菌与饮食诱导的肥胖密切相关,为进一步研究SRB在肥胖及其相关代谢疾病的发生发展中的作用提供了依据。

Objective To quantify the numerical and proportional changes of sulfate-reducing bacteria（SRB） in the gut of rats with high-fat diet（HFD） intervention.Methods 20 Wistar rats were assigned to two groups（n=10 for each group） randomly.One group of rats was fed with HFD for 18 weeks and the other group（control） was fed with normal chow diet（NCD）.Real-time qPCR was employed to assess the SRB levels in both groups at 0,8 and 18 weeks respectively by determining the number of copies of the marker gene aprA.The total bacteria was also quantified targeted 16S rRNA gene,to evaluate the dynamic changes of the proportion of SRB in the rat gut microbial community.Results After 8 weeks feeding,the rats in HFD group gained significantly more body weight than NCD group.Meanwhile,SRB in the gut microbiota were significantly enriched in this group compared with the NCD group.Conclusion Our study results suggests a close relationship between SRB and diet-induced obesity,and provides the fundamental support to further study on the functional role of SRB in the development of obesity and related metabolic diseases.