摘要
目的探究小分子RNA干扰受体型蛋白酪氨酸磷酸酶样O(PTPRO)表达对克罗恩病(CD)大鼠的作用及机制。方法将40只大鼠随机分为4组:对照组(A组)、模型组(B组,TNBS+尾静脉注射生理盐水)、空载体组(C组,TNBS+尾静脉注射含空载质粒的慢病毒)和干扰组(D组,TNBS+尾静脉注射含si-PTPRO mRNA质粒的慢病毒),每组各10只。采用TNBS乙醇溶液灌肠构建CD大鼠模型。采用HE染色方法观察肠黏膜组织的病理学变化,实时荧光定量PCR法检测PTPRO mRNA水平,酶联免疫法检测肠黏膜组织中白细胞介素-1β(IL-1β)、IL-6、IL-10及肿瘤坏死因子-α(TNF-α)水平,蛋白质免疫印迹法检测PTPRO、Toll样受体4(TLR4)及核因子κB(NF-κB)蛋白水平。结果与A组比较,B组的CD疾病活动指数、PTPRO mRNA、IL-1β、IL-6、TNF-α、PTPRO、TLR4及NF-κB水平均升高,IL-10水平降低,差异均有统计学意义(P均<0.05)。与B组和C组比较,D组的CD疾病活动指数、PTPRO mRNA、IL-1β、IL-6、TNF-α、PTPRO、TLR4及NF-κB水平均降低,IL-10水平升高,差异均有统计学意义(P均<0.05)。结论抑制PTPRO表达对CD大鼠有一定保护作用,其机制可能与调控TLR4、NF-κB表达有关。
Objective This paper aims to investigate the effect and mechanism of small molecule RNA interference protein tyrosine phosphatase receptor-type O(PTPRO)expression on rats with Crohn′s disease(CD).Methods Forty rats were randomly divided into the control group(Group A),the model group(Group B,TNBS+tail vein injection of saline),the empty vector group(Group C,TNBS+tail vein injection of lentivirus containing empty plasmid),and the interference group(Group D,TNBS+tail vein injection of lentivirus containing si-PTPRO mRNA plasmid),with 10 rats in each group.A rat model of CD was constructed with TNBS ethanol solution enema.HE staining was used to observe the pathological changes of intestinal mucosa tissue;real-time fluorescent quantitative PCR was used to detect the level of PTPRO mRNA;enzyme-linked immunosorbent assay was used to detect the levels of interleukin-1β(IL-1β),IL-6,IL-10,and tumor necrosis factorα(TNF-α)in intestinal mucosa;Western blot was used to detect PTPRO,TLR4,and NF-κB protein levels.Results Compared with Group A,the levels of CD activity index,PTPRO mRNA,IL-1β,IL-6,TNF-α,PTPRO,TLR4,and NF-κB in Group B increased,while the level of IL-10 decreased(P<0.05).Compared with Groups B and C,the levels of CD activity index,PTPRO mRNA,IL-1β,IL-6,TNF-α,PTPRO,TLR4,and NF-κB in Group D decreased,while the level of IL-10 increased(P<0.05).Conclusion Inhibition of PTPRO expression has a protective effect on rats with CD,and the mechanism may be related to the regulation of TLR4 and NF-κB expression.
作者
张强
丛中笑
ZHANG Qiang;CONG Zhongxiao(Department of Gastroenterology,Shandong Provincial Third Hospital,Jinan 250000,China;Department of Operation room,Qilu Hospital of Shandong University,Jinan 250012,China)
出处
《国际消化病杂志》
CAS
2020年第3期171-174,共4页
International Journal of Digestive Diseases
