左氨氯地平对慢性肾衰竭大鼠肠道损伤的保护作用及肠道菌群失调的影响

Protective Effect of Levamlodipine on Intestinal Injury and Dysbiosis of Intestinal Flora in Rats with Chronic Renal Failure

目的:探索左氨氯地平对慢性肾衰竭大鼠模型的肠道菌群失调和肠道损伤的影响。方法:SDS大鼠19只,饲养于SPF级环境,对照组5只,5/6肾脏切除组6只,5/6肾脏切除+左氨氯地平干预8只。大鼠于造模后2周至10周予左氨氯地平3 mg·kg^(-1)·d^(-1)灌胃处理,10周处死大鼠。于造模后4周和10周测量血压。处死前取大鼠粪便,进行16S核糖体DNA(rDNA)高通量测序;4周和处死时留取血液,行肾功能等生化检测;处死后取肾脏组织、肠道组织行病理染色观察结构改变和免疫组化染色检查。结果:与对照组相比,5/6肾脏切除组大鼠10周的收缩压升高,而与5/6肾脏切除组相比,左氨氯地平干预后可降低收缩压。在4周和10周时,与对照组相比,5/6肾脏切除组血肌酐和血尿素氮明显升高;而与5/6肾脏切除组相比,在4周和10周时左氨氯地平干预组血肌酐和尿素氮水平无明显差别。在HE染色水平,与5/6肾脏切除组比较,左氨氯地平可减轻肾脏损伤程度和肠道炎症、水肿。在回肠组织中,与5/6肾脏切除组相比,左氨氯地平可改善模型引起的肠道连接蛋白occludin和ZO-1的表达下降。在肠道菌群方面,左氨氯地平干预改变5/6肾脏切除组大鼠的肠道微生物的α多样性、β多样性,且左氨氯地平干预可改善5/6肾脏切除组大鼠的肠道微生物的多样性。在属水平,左氨氯地平干预可改变5/6肾脏切除组大鼠肠道的拟杆菌属、毛螺菌属、普氏菌属、颤螺菌属、梭菌属_UCG-014、颤螺旋菌科、罗姆布茨菌、Allobaculum、Colidextribacter、副拟杆菌、罗氏菌属、普雷沃菌科_Ga6A1组、肠球菌、Fournierella的水平。结论:左氨氯地平一定程度改善5/6肾脏切除组大鼠肾脏损伤和肠道损伤以及改变肠道菌群,其对肠道和肠道菌群的作用与肾脏的保护作用的关联需进一步研究。

Objective:To explore the effect of levamlodipine on dysbiosis of intestinal flora and intestinal injury in a rat model of renal fibrosis.Methods:Nineteen SDS rats were housed in SPF class environment(five in the 5/6 nephrectomy group control group,six in the 5/6 nephrectomy group,and eight in the 5/6 nephrectomy+levamlodipine group).The rats were treated with levamlodipine 3 mg/kg/d by gavage from 2 weeks to 10 weeks after modeling,and the rats were executed at 10 weeks.Blood pressure was measured at 4-weeks and 10-weeks post-modeling.Blood was collected at 4 weeks and at the time of execution for biochemical tests including renal function.After execution,kidney and intestinal tissues were stained for structural changes and immunohistochemical staining.Results:Compared with the model group,systolic blood pressure was increased at 10 weeks after modeling in rats,while levamlodipine intervention reduced systolic blood pressure.At 4 and 10 weeks,blood creatinine and blood urea nitrogen were significantly elevated in the 5/6 nephrectomy group compared with the control group,while there was no significant difference in blood creatinine and urea nitrogen levels in the levamlodipine intervention group at 4 and 10 weeks compared with the 5/6 nephrectomy model group.Also,evamlodipine reduced the degree of kidney injury and intestinal inflammation and edema compared with the model group.In ileal tissue,levamlodipine increased the expression of intestinal linker proteins occludin and ZO-1 compared with the model group.In terms of intestinal flora,levamlodipine intervention altered the alpha diversity,beta diversity of intestinal microbes in 5/6 nephrectomized rats,and levamlodipine intervention improved the diversity of intestinal microbes in 5/6 nephrectomized rats.At the genus level,levamlodipine altered the intestinal microbial diversity of the 5/6 nephrectomized rats in Bacteroides,Lachnospiraceae,Prevotella,Oscillospiraceae,Clostridia_UCG-014,Oscillospiraceae,Romboutsia,Allobaculum,Colidextribacter,Parabacteroides,Roseburia,Prevotellaceae_Ga6A1_group,Enterococcus,and Fournierella levels.Conclusion:Levamlodipine ameliorated renal injury and intestinal injury as well as altered intestinal flora in the 5/6 nephrectomy model,and the association between its effects on the intestine and intestinal flora and the protective effect on the kidney needs further study.