车前子多糖对膜性肾病大鼠肾损伤和肠道菌群的影响  

Effect of Polysaccharides from Plantaginis Semen on Renal Injury and Gut Microbiota in Rats with Membranous Nephropathy

作　　者：赵宏[1] 陈晨 赵岩[1] 汤威威 高琪 孔令洲 于登君 张宇[1] ZHAO Hong;CHEN Chen;ZHAO Yan;TANG Wei-wei;GAO Qi;KONG Ling-zhou;YU Deng-jun;ZHANG Yu(College of Pharmacy,Jiamusi University,Jiamusi 154007,China)

机构地区：[1]佳木斯大学药学院,黑龙江佳木斯154007

出　　处：《中国实验方剂学杂志》2021年第22期92-99,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中央支持地方高校改革发展资金人才培养支持计划项目(2019zyzcdf-01);黑龙江省博士后专项经费项目(LBH-Q20185);黑龙江省北药与功能食品特色学科建设项目(2018-TSXK-02);佳木斯大学优秀学科团队项目(JDXKTD-2019005)。

摘　　要：目的:研究车前子多糖(PSP)对膜性肾病(MN)大鼠肾损伤的保护作用及对肠道菌群的影响,为PSP治疗MN的深入研究提供理论基础。方法:采用尾静脉注射阳离子化牛血清白蛋白(C-BSA,3.5 g·L^(-1))诱导MN大鼠模型,造模周期为7周。于造模第4周,将造模成功大鼠按照随机数字法分为模型组,阳性药物组(盐酸贝那普利,10 mg·kg^(-1)·d^(-1)),PSP高剂量组(PSP-H,800 mg·kg^(-1)·d^(-1)),PSP中剂量组(PSP-M,400 mg·kg^(-1)·d^(-1)),PSP低剂量组(PSP-L,200 mg·kg^(-1)·d^(-1)),每组10只;另取10只未造模的大鼠为正常组。正常组和模型组大鼠灌服等体积生理盐水,各给药组大鼠灌服相应药物,每日1次,连续4周。实验结束后,采用光镜法分别观察大鼠肾脏及结肠组织病理变化,酶联免疫吸附测定法(ELISA)检测大鼠血清及结肠组织中肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL^(-1)β)含量,免疫组织化学法检测大鼠肾组织中TNF-α,IL^(-1)β蛋白表达情况,运用16S r RNA测序法研究PSP对MN大鼠肠道菌群调节作用。结果:与正常组比较,模型组大鼠肾小球增大、基底膜增厚,结肠组织腺体萎缩,血清及结肠组织中TNF-α,IL^(-1)β含量明显上升(P<0.05),TNF-α,IL^(-1)β蛋白表达显著增加(P<0.01);与模型组比较,盐酸贝那普利组和PSP-H组大鼠肾小球囊腔减小、基膜增厚程度减轻、结肠组织中黏膜排列较整齐,但PSP-M,PSP-L组对肾组织及结肠组织改善效果不佳;PSP-H,PSP-M组大鼠血清及结肠组织中TNF-α,IL^(-1)β含量明显下降(P<0.05),肾组织中TNF-α,IL^(-1)β蛋白表达显著降低(P<0.01),PSP-L组差异无统计学意义;模型组大鼠肠道菌群中有害菌厚壁菌门丰度升高、有益菌拟杆菌门丰度降低;给予PSP后有害菌厚壁菌门丰度降低、有益菌拟杆菌门丰度升高,以PSP-H变化最为显著。结论:PSP可有效缓解MN大鼠的肾损伤、降低炎症因子表达,并可调节MN大鼠肠道菌群结构、改善受损的肠Objective:To study the protective effect of polysaccharides from Plantaginis Semen(PSP)against renal injury in rats with membranous nephropathy(MN)and its influence on the gut microbiota to provide a theoretical basis for the further investigation of PSP in the treatment of MN. Method: The MN model was induced by tail vein injection of cationic bovine serum albumin(C-BSA,3.5 g·L^(-1))in rats with a modeling period of seven weeks. At the 4 th week of modeling,the model rats were divided into a model group,a positive drug group(benazepril hydrochloride,10 mg·kg^(-1)·d^(-1)),a PSP high-dose group(PSP-H,800 mg·kg^(-1)·d^(-1)),a PSP medium-dose group(PSP-M,400 mg·kg^(-1)·d^(-1)),and a PSP low-dose group(PSP-L,200 mg·kg^(-1)·d^(-1))according to the random number table,with 10 in each group. Ten healthy rats were assigned to the normal control group. The rats in the normal control group and the control group received an equal amount of physiological saline by gavage, and those in the groups with drug intervention were administered correspondingly,once a day,for consecutive four weeks. The pathological changes of rat kidney and colon tissues were observed by optical microscopy. The enzyme-linked immunosorbent assay(ELISA)was used to detect the content of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL^(-1)β)in the serum and colon tissues.The immunohistochemistry(IHC)was used to detect the protein expression of TNF-α and IL^(-1)β in renal tissues.The 16 S rRNA sequencing method was used to investigate the effect of PSP on the gut microbiota in MN rats.Result: Compared with the normal control group,the model group showed enlarged glomeruli,thickened basement membrane,atrophied colonic gland,increased TNF-α and IL^(-1)β in the serum and colon tissues(P<0.05),and elevated protein expression of TNF-α and IL^(-1)β(P<0.01). Compared with the model group,the positive drug group and the PSP-H group displayed shrunk glomerular capsules,relieved basement membrane thickening,and neatly arranged colonic mucosa

关 键 词：车前子 多糖 膜性肾病 肾损伤 肠道菌群 

分 类 号：R2-0[医药卫生—中医学] R22