CAJ | 학술논문

目的：观察和寻找慢性温和不可预知应激程序( CUMS)抑郁模型药物干预后的菌群变化并鉴定具体菌种。方法SD大鼠随机分为空白对照组、模型组、盐酸氟西汀组和逍遥散组；采用慢性温和不可预知应激程序( CUMS)复制大鼠抑郁模型,在第1天时除空白组和模型组给予等体积生理盐水外,其他两组分别给予逍遥散和盐酸氟西汀,28 d后取盲肠；提取盲肠基因组DNA,并对16 S rRNA基因V3可变区进行PCR扩增,构建变性梯度凝胶电泳( DGGE)指纹图谱并制定碱基序列。将PCR扩增产物与大肠杆菌DH5α感受态细胞混合培养,选取形成的白色菌落,进行鉴定分析。结果体重和行为学指标显示CUMS复制成功；盲肠指数显示逍遥散可回调慢性应激对盲肠所产生的影响。采用BLAST程序对盲肠菌群DGGE指纹图谱及测得的序列进行相似性分析发现Lachnospiraceae bacterium、Lactobacillus animali、Burkholderiales bacterium、Lactobacillus reu-teri四种优势菌。与空白对照组相比,Lachnospiraceae bacterium、Lactobacillus animali、Burkholderiales bacterium三种菌种的丰度在模型组均增强,Lactobacillus reuteri在模型组显著减弱。给予逍遥散后,Lactobacillus animali的丰度增强,Lachnospiraceae bac-terium的丰度不变,而其他两种细菌未检测到。结论通过DGGE分析发现抑郁症大鼠的盲肠菌群发生明显改变,逍遥散通过调节抑郁症大鼠肠道菌群的益生菌来改善胃肠功能。
목적：관찰화심조만성온화불가예지응격정서( CUMS)억욱모형약물간예후적균군변화병감정구체균충。방법SD대서수궤분위공백대조조、모형조、염산불서정조화소요산조；채용만성온화불가예지응격정서( CUMS)복제대서억욱모형,재제1천시제공백조화모형조급여등체적생리염수외,기타량조분별급여소요산화염산불서정,28 d후취맹장；제취맹장기인조DNA,병대16 S rRNA기인V3가변구진행PCR확증,구건변성제도응효전영( DGGE)지문도보병제정감기서렬。장PCR확증산물여대장간균DH5α감수태세포혼합배양,선취형성적백색균락,진행감정분석。결과체중화행위학지표현시CUMS복제성공；맹장지수현시소요산가회조만성응격대맹장소산생적영향。채용BLAST정서대맹장균군DGGE지문도보급측득적서렬진행상사성분석발현Lachnospiraceae bacterium、Lactobacillus animali、Burkholderiales bacterium、Lactobacillus reu-teri사충우세균。여공백대조조상비,Lachnospiraceae bacterium、Lactobacillus animali、Burkholderiales bacterium삼충균충적봉도재모형조균증강,Lactobacillus reuteri재모형조현저감약。급여소요산후,Lactobacillus animali적봉도증강,Lachnospiraceae bac-terium적봉도불변,이기타량충세균미검측도。결론통과DGGE분석발현억욱증대서적맹장균군발생명현개변,소요산통과조절억욱증대서장도균군적익생균래개선위장공능。
Objective To observe and identify the flora changes in chronic unpredictable mild stress( CUMS) model after intervened by drugs. Methods All SD rats were randomized into four groups:healthy control group, model group,Xiaoyaosan group and fluoxer-tine hydrochloride group. The depressive rat model was coupled by chronic unpredictable mild stress( CUMS) . The rats in healthy con-trol group and model group were given volumetric saline,while the rats in the other groups were given Xiaoyaosan and fluoxetine hydro-chloride at 1 d,respectively. The cecum was taken at 28 d and the genomic DNAs were extracted from all of samples. Denaturing gradi-ent gel electrophoresis(DGGE)fingerprints were constructed by PCR amplification of 16S rRNA V3 variable region. The PCR amplifica-tion product was mixed with competent cells of Escherichia coli DH5α, and formed white colonies were selected and identified. Re-sults The results of body weight and behavior indicated the CUMS model was successfully established, and the index of cecum showed that Xiaoyaosan improved the impact of caecum stimulated by chronic unpredictable mild stress. Combined with DGGE fingerprint of the cecum flora and the measured sequence,the analysis of similarity by the BLAST program showed that four kinds of dominant bacteria were screened, including Lachnospiraceae bacterium, Lactobacillus animali, Burkholderiales bacterium and Lactobacillus reuteri. Com-pared with normal control group,the abundance of Lachnospiraceae bacterium,Lactobacillus animali and Burkholderiales bacterium was enhanced in CUMS group, and the abundance of Lactobacillus reuteri was weakened. After administration of Xiaoyaosan, the abun-dance of Lactobacillus animali was enhanced, the abundance of Lachnospiraceae bacterium showed no significant change and others were not found. Conclusion The cecum flora of depressive rats change markedly by the analysis of DGGE. Xiaoyaosan can regulate the probiotics in cecum flora of depressive rats to improve the gastrointestinal disorder.