右美托咪定对腹腔镜气腹大鼠肠屏障功能及肠道菌群的影响OA北大核心CSTPCD
Effect of Dexmedetomidine on Intestinal Barrier Function and Intestinal Microbiota in Laparoscopic Pneumoperitoneum Rats
[目的]探究右美托咪定(dexmedetomidine,DEX)对腹腔镜气腹大鼠肠屏障功能及肠道菌群的影响.[方法]试验将30只SD大鼠随机分为3组:假手术组(Sham)(n=6)、CO2气腹组(CO2)(n=12)、DEX组(n=12).CO2和DEX组大鼠在气腹前30 min分别腹腔注射1 mL/kg生理盐水和50 μg/kg右美托咪定,随后在15 mmHg气腹压力下维持90 min建立气腹模型.Sham组不通气腹,其余操作同CO2组.气腹结束后12、24 h采集大鼠回肠组织,观察其组织病理形态学变化,并通过试剂盒测定大鼠回肠组织中丙二醛(MDA)、谷胱甘肽(GSH)、白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子-α(TNF-α)含量以及髓过氧化物酶(MPO)活性;利用实时荧光定量PCR检测闭锁小带蛋白-1(ZO-1)、封闭蛋白1(Claudin-1)、闭合蛋白(Occludin)的mRNA相对表达量;采集回肠组织内容物,通过高通量双端测序检测大鼠回肠菌群的变化.[结果]与Sham组相比,CO2组大鼠回肠组织结构紊乱,肠绒毛变形,出现炎性细胞浸润等现象;结肠中MDA、IL-6、IL-1β、TNF-α含量及MPO活性均极显著升高(P<0.01),GSH含量以及ZO-1、Occludin、Claudin-1 基因mRNA表达量均极显著降低(P<0.01);肠道菌群多样性减少,门水平上,厚壁菌门(Firmicutes)相对丰度有降低趋势(P>0.05),放线菌门(Actinobacteria)、变形菌门(Proteobacteria)相对丰度有增加趋势(P>0.05).属水平上,乳杆菌属(Lactobacillus)和异杆菌属(Allobaculum)相对丰度有降低趋势(P>0.05),而双歧杆菌属(Bifidobacterium)相对丰度有增加趋势(P>0.05).与CO2组相比,DEX组大鼠回肠组织结构紊乱及肠绒毛变形情况缓解,炎性细胞浸润减少.结肠中GSH含量及ZO-1、Claudin-1、Occludin基因mRNA表达量均极显著或显著升高(P<0.01;P<0.05),MDA、IL-6、IL-1β、TNF-α含量及MPO活性均显著或极显著降低(P<0.05;P<0.01);肠道菌群多样性增加,门水平上,厚壁菌门相对丰度有增加趋势(P>0.05),放线菌门、变形菌门相对丰度有降低趋势(P>0.05).属水平上,乳杆菌属相对丰度有增加趋势(P>0.05),双歧杆菌属和异杆菌属相对丰度有降低趋势(P>0.05).[结论]DEX可以通过改善机体氧化应激、炎症因子的释放、肠道菌群结构等,增加肠道菌群丰度及多样性,从而缓解腹腔镜气腹所致的肠损伤.
[Objective]The purpose of this study was to investigate the effects of dexmedetomidine(DEX)on intestinal barrier function and intestinal microbiota in rats undergoing laparoscopic pneumoperitoneum.[Method]Thirty SD rats were randomly divided into three groups:Sham group(n=6),CO2 pneumoperitoneum group(CO2)(n=12)and DEX group(n=12).The rats in CO2 and DEX groups were intraperitoneally injected with 1 mL/kg normal saline and 50 μg/kg DEX 30 min before pneumoperitoneum,respectively,and then maintained at 15 mmHg pneumoperitoneum pressure for 90 min to establish an pneumoperitoneum model.Pneumoperitoneum was impassable in Sham group,and other operations were the same as those in CO2 group.The ileum tissues were collected 12 and 24 h after pneumoperitoneum,and the histopathologic changes were observed.The contents of malondialdehyde(MDA),glutathione(GSH),interleukin-6(IL-6),IL-1β,tumor necrosis factor-α(TNF-α)and myeloperoxidase(MPO)activity of ileum in rats were determined by the kit.The mRNA relative expression of occlusive zonal-1(ZO-1),occlusive protein-1(Claudin-1)and occlusive protein(Occludin)was detected by Real-time quantitative PCR.The contents of ileal tissue were collected and the changes of ileal microflora were detected by high-throughput paired-end sequencing.[Result]Compared with Sham group,the structure of ileum was disordered,intestinal villi deformed and inflammatory cell infiltrated in CO2 group.MDA,IL-6,IL-1β and TNF-α contents,and MPO activity were extremely significantly increased(P<0.01),GSH content and mRNA expression of ZO-1,Occludin,Claudin-1 genes were extremely significantly decreased(P<0.01).The diversity of intestinal flora decreased.At the phylum level,the relative abundance of Firmicutes showed a decreasing trend(P>0.05),while the relative abundance of Actinobacteria and Proteobacteria showed an increasing trend(P>0.05).At the genus level,the relative abundance of Lactobacillus and Allobaculum showed a decreasing trend(P>0.05),while the relative abundance of Bifidobacterium showed an increasing trend(P>0.05).Compared with CO2 group,the ileal tissue structure disorder and intestinal villi deformation of rats in DEX group were relieved,and inflammatory cell infiltration was reduced.GSH content and mRNA expression of ZO-1,Claudin-1 and Occludin genes were extremely significantly or significantly increased(P<0.01 or P<0.05),MDA,IL-6,IL-1β and TNF-α contents,and MPO activity were significantly or extremely significantly decreased(P<0.05 or P<0.01).The diversity of intestinal flora increased.At the phylum level,the relative abundance of Firmicutes showed an increasing trend(P>0.05),while the relative abundance of Actinobacteria and Proteobacteria showed a decreasing trend(P>0.05).At the genus level,the abundance of Lactobacillus showed an increasing trend(P>0.05),while the abundance of Bifidobacterium and Allobaculum showed a decreasing trend(P>0.05).[Conclusion]DEX could increase the abundance and diversity of intestinal microbiota by improving oxidative stress,the release of inflammatory factors,and the structure of intestinal flora,so as to alleviate intestinal damage caused by laparoscopic pneumoperitoneum.
宁夏青;王会英;张暖暖;王佳琦;李雪琴;蒋聪聪;翟帆;张士霞
河北农业大学动物医学院,保定 071000安阳市动物疫病预防控制中心,安阳 455000
畜牧业
右美托咪定CO2气腹氧化应激肠屏障肠道菌群
dexmetomidineCO2 pneumoperitoneumoxidative stressintestinal barrierintestinal microbiota
《中国畜牧兽医》 2024 (007)
3176-3184 / 9
国家自然科学基金(31802250);河北省教育厅科学研究项目(BJ2019053、QN2015184);河北省自然科学基金(C2023204085)
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