基于靶向代谢组学探讨葛根芩连汤对抗生素相关性腹泻大鼠粪便短链脂肪酸的影响

Effect of Gegen Qinliantang on Fecal Short-chain Fatty Acids in Rats with Antibiotic-associated Diarrhea Based on Targeted Metabonomics

目的:通过靶向代谢组学探究葛根芩连汤对抗生素相关性腹泻(AAD)粪便短链脂肪酸(SCFAs)代谢的影响。方法:将240只SD大鼠随机分为空白组、模型组、丽珠肠乐组(0.15 g·kg^(-1))、葛根芩连汤高、中、低剂量组(10.08、5.04、2.52 g·kg^(-1)),每组40只(雌雄各半),除空白组外,其余各组每日均给予克林霉素(250 mg·kg^(-1))灌胃造模,连续7 d。造模成功后,各给药组按相应剂量灌胃药液,空白组和模型组灌胃等体积生理盐水,1次/d,连续治疗14 d。分别在药物干预后0、3、7、14 d,从各组中随机抽取8只大鼠,气相色谱-飞行时间质谱法(GC-TOF-MS)对大鼠粪便中SCFAs进行靶向代谢组学分析,运用偏最小二乘法-判别分析(PLS-DA)比较不同治疗时间组间代谢轮廓差异,并对大鼠粪便中SCFAs的组间含量变化进行比较。结果:PLS-DA结果显示,空白组与模型组可明显区分,在葛根芩连汤治疗7 d后逐渐趋近于空白组。进一步分析SCFAs组成后发现,与空白组比较,模型组中乙酸的占比增加,丁酸、戊酸、己酸和异戊酸的占比降低;葛根芩连汤治疗后,丁酸、异丁酸、戊酸和异戊酸占比增加,乙酸、丙酸和己酸占比降低。经差异分析显示,葛根芩连汤可明显改善丁酸含量,对戊酸、己酸含量具有一定回调作用。结论:葛根芩连汤中剂量组治疗7 d后可改善AAD粪便短链脂肪酸含量,这可能与改善SCFAs的组成比例及丁酸、戊酸、己酸等含量有关。

Objective:To explore the impact of Gegen Qinliantang(GQT) on the fecal short-chain fatty acids(SCFAs) metabolism in antibiotic-associated diarrhea(AAD) through targeted metabolomics.Method:A total of 240 SD rats were randomly divided into six groups(n=40,half male and half female),including blank group,model group,bifidobiogen group(0.15 g·kg^(-1)),and GQT high-,medium-,and low-dose groups(10.08,5.04,2.52 g·kg^(-1)),except for the blank group,clindamycin(250 mg·kg^(-1)) was given to all groups by gavage for modeling every day for 7 d.After successful modeling,each administered group was gavaged with the corresponding dose of the drug,and the blank and model groups were gavaged with an equal volume of normal saline solution,1 time/d,for 14 d.At 0,3,7,14 d after the drug intervention,eight rats were randomly selected from each group,respectively.Gas chromatography-time-of-flight mass spectrometry(GC-TOF-MS) was used to perform targeted metabolomic analysis of SCFAs in the feces of rats,and partial least squaresdiscriminant analysis(PLS-DA) was applied to compare the differences in metabolic profiles between groups at different treatment times,and to compare the changes in the contents of SCFAs in rat feces between groups.Result:PLS-DA results showed that the blank group could be clearly distinguishable from the model group,with GQT exhibiting a closer proximity to the blank group after 7 d of treatment.After further analyzing the composition of SCFAs,it was found that the proportion of acetic acid increased and the proportions of butyric acid,valeric acid,hexanoic acid and isovaleric acid decreased in the model group compared with the blank group.After the treatment with GQT,the proportions of butyric acid,isobutyric acid,valeric acid,and isovaleric acid increased,and the proportions of acetic acid,propionic acid and caproic acid decreased.Subsequent differential analysis revealed that GQT could significantly improve the content of butyric acid,and had a certain retrogressive effect on the contents of valeric acid and hexanoic acid.Conclusion:The medium dose group of GQT can improve the contents of SCFAs in AAD feces after 7 days of treatment,which may be related to the improvement of the composition ratio of SCFAs and the contents of butyric acid,valeric acid and caproic acid.