基于肠道菌群-宿主代谢相关性的大枣降低十枣汤有毒药味肠道损伤作用研究  

Jujubae Fructus alleviates intestinal injury caused by toxic medicinals in Shizao Decoction based on correlation between intestinal flora and host metabolism

作　　者：高小琴 徐金娣[2] 周世康 张毅[2] 张丽 GAO Xiao-qin;XU Jin-di;ZHOU Shi-kang;ZHANG Yi;ZHANG Li(Hanlin College,Nanjing University of Chinese Medicine,Nanjing 225300,China;Jiangsu Provincial Collaborative Innovation Center for Industrialization Process of Traditional Chinese Medicine Resources/National-Local Joint Engineering Research Center for Traditional Chinese Medicine Resources Industrialization,Prescriptions and Innovative Drugs,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学翰林学院,江苏泰州225300 [2]南京中医药大学江苏省中药资源产业化过程协同创新中心/中药资源产业化与方剂与创新药物国家地方联合工程研究中心,江苏南京210023

出　　处：《中国中药杂志》2023年第10期2792-2802,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81673599);江苏省自然科学基金项目(BK20201232);2020年度省第五期“333高层次人才培养工程”项目(BRA20200071)。

摘　　要：十枣汤中有毒药味甘遂、大戟、芫花对肠道有明显毒性,组方时虽采用大枣降低上述有毒药味的毒性,但其机制尚欠清晰。为探索大枣对十枣汤组方有毒药味肠道毒性的减毒作用机制,选择正常SD大鼠40只,分为空白组,十枣汤高、低剂量组,十枣汤有毒药味高、低剂量组共5组,分别灌胃给予加枣汤和不加枣汤的方药,采用体质量变化、脾脏指数表征体质状态;苏木精-伊红(HE)染色法观察肠组织病理学变化,测定小肠组织中丙二醛(MDA)和谷胱甘肽(GSH)含量以及超氧化物歧化酶(SOD)活力以评价肠道损伤;收集新鲜粪便采用16S核糖体RNA基因(16S rDNA)测序技术检测肠道菌群结构变化;气相色谱质谱联用仪(GC-MS)定量分析粪便短链脂肪酸的含量;快速液相色谱串联四极杆飞行时间质谱(UFLC-Q-TOF-MS)技术测定粪便代谢物,并用Spearman关联网络分析差异性菌属和差异代谢产物。结果显示,与空白组相比,十枣汤有毒药味高、低2个剂量组大鼠小肠组织中MDA含量显著增加,GSH含量和SOD活力显著降低,各肠段肠绒毛显著缩短(P<0.05);肠道菌群多样性和丰富度降低,菌群结构改变;短链脂肪酸含量显著降低(P<0.05)。与十枣汤有毒药味各剂量组相比,十枣汤高、低剂量组大鼠小肠组织中MDA含量显著降低,GSH含量和SOD活力显著升高(P<0.05),肠绒毛长度回调;肠道菌群多样性和丰富度有所增加,肠道菌群失调的状态改善;短链脂肪酸含量显著回调(P<0.05)。结合大枣配伍后肠道菌群和粪便代谢产物变化,筛选出乳杆菌属、丁酸单胞菌属、Clostridia_UCG-014菌属、普雷沃菌属、大肠杆菌-志贺菌属和另枝杆菌属6种差异菌属,乙酸、丙酸、丁酸、戊酸4种短链脂肪酸和尿石素A、石胆酸、肌酸酐等18种差异代谢物。关联分析发现,有益菌属乳酸杆菌等与粪便代谢产物丁酸和尿石素A等显著正相关(P<0.05);致病菌属大肠杆菌-志贺�Genkwa Fols,Kansui Radix,and Euphorbiae Pekinensis Radix in Shizao Decoction(SZD)are toxic to intestinal tract.Jujubae Fructus in this prescription can alleviate the toxicity,but the mechanism is still unclear.Therefore,this s tudy aims to explore the mechanism.To be specific,40 normal Sprague-Dawley(SD)rats were classified into the normal group,high-dose and low-dose SZD groups,and high-dose and low-dose SZD without Jujubae Fructus(SZD-JF)groups.The SZD groups were given(ig)SZD,while SZD-JF groups received the decoction without Jujubae Fructus.The variation of body weight and spleen index were recorded.The pathological changes of intestinal tissue were observed based on hematoxylin and eosin(HE)staining.The content of malondialdehyde(MDA)and glutathione(GSH)and activity of superoxide dismutase(SOD)in intestinal tissue were measured to evaluate the intestinal injury.Fresh feces of rats were collected to detect inte stinal flora structure by 16S ribosomal RNA gene(16S rDNA)sequencing technology.The content of fecal short chain fatty acids and fecal metabolites was determined by gas chromatography-mass spectrometer(GC-MS)and liquid chromatography-mass spectrometer ultra-fast liquid chromatography-quadrupole-time-of-flight mass spectrometer(UFLC-Q-TOF-MS),separately.Spearman’s correlation analysis was employed to analyze the differential bacteria genera and differential metabolites.Result s showed that high-dose and low-dose SZD-JF groups had high content of MDA in intestinal tissue,low GSH content and SOD activity,short intestinal villi(P<0.05),low diversity and abundance of intestinal flora,variation in the intestinal flora structure,and low content of short chain fatty acids(P<0.05)compared with the normal group.Compared with high-dose and low-dose SZD-JF groups,high-dose and low-dose SZD groups displayed low content of MDA in intestinal tissue,high GSH content and SOD activity,recovery of the length of intestinal villi,increased abundance and diversity of intestinal flora,alleviation of dysbacteria,and recover

关 键 词：十枣汤 肠道损伤 肠道菌群 短链脂肪酸 代谢组学 

分 类 号：R285.5[医药卫生—中药学]