重组人乳铁蛋白对幽门螺旋杆菌胃炎小鼠空泡毒素蛋白A、肿瘤坏死因子α含量的影响

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目的:本研究探讨重组人乳铁蛋白(recombinant human lactoferrin,rhLF)联合三联疗法是否能够提高小鼠胃幽门螺旋杆菌(Helicobacter pylori,H.pylori)根除率,能否降低小鼠胃黏膜炎症反应,初步揭示rhLF对H.pylori感染小鼠胃黏膜炎症的作用特点及相关机制.方法:192只Babl/c小鼠用H.pylori ATCC43504感染建立胃炎模型,随机分为A组(标准三联+rhLF)、B组(rhLF)、C组(标准三联)、D组(生理盐水),小鼠胃黏膜采用特殊银染评定H.pylori定植情况、HE染色镜检炎症积分值,采用ELISA试剂盒测定胃组织匀浆肿瘤坏死因子(tumor necrosis factor,TNF)-α含量.并通过RT-PCR方法检测各组药物对H.pyloriATCC43504的主要致病因子空泡毒素蛋白(vacuolating cytotoxin,VacA)mRNA表达水平.数据均有SPSS17.0软件分析.结果:与D组比较,A、B、C组小鼠胃黏膜特殊银染色镜检H.pylori定植率、HE染色镜检胃黏膜炎症积分明显降低(P<0.05);与B、C组比较,A组小鼠胃黏膜特殊银染色镜检H.pylori定植、HE染色镜检胃黏膜炎症积分明显降低(P<0.05);A、B、C、D组胃组织匀浆中体内TNF-α含量分别为28.64pg/mL±12.07 pg/mL、300.16 pg/mL±59.10pg/mL、54.96 pg/mL±15.02 pg/mL、503.25pg/mL±1.35 pg/mL;rhLF联合三联疗法可降低胃组织匀浆液中TNF-α含量(P<0.01),与生理盐水组、标准三联组、rhLF组比较差异有统计意义.rhLF联合三联疗法可降低胃组织匀浆液中VacA mRNA表达水平(P<0.01),与生理盐水组、标准三联组、rhLF组比较差异有统计意义.结论:RhLF联合三联疗法能够提高H.pylori根除率;降低胃黏膜炎症反应,此作用可能与rhLF联合三联疗法能够降低促炎症因子释放有关.rhLF可有效抑制VacA mRNA表达,提示rhLF具有下调H.pylori毒性的潜在作用. Objective: This study was designed to investigate whether recombinant human lactoferrin (rhLF) combined with triple therapy can improve the eradication rate of Helicobacter pylori (H. pylori) in mice and reduce the gastric mucosal inflammation in mice, To investigate the effect and mechanism of rhLF on gastric mucosal inflammation in mice infected with H.pylori.Methods: 192 Babl / c mice were infected with H.pylori ATCC43504 to establish gastritis model and were randomly divided into group A (standard triple combined with rhLF) , Group B (rhLF), group C (standard triple) and group D (normal saline). The gastric mucosa of mice were used to evaluate the H.pylori colonization by special silver staining. The inflammation score was examined by HE staining. The content of tumor necrosis factor (TNF) -α was detected by enzyme-linked immunosorbent assay (ELISA). The expression of vacuolating cytotoxin (VacA) mRNA in H.pylori ATCC 43504 was detected by RT-PCR. SPSS17.0 software was used to analyze the data.Results: Compared with group D, H.pylori colonization rate of gastric mucosa of mice in group A, B and C was significantly higher than that in group D. Gastric mucosal inflammation score was significantly decreased by HE staining (P < 0.05); Compared with B, C group, A group Gastric mucosa of mice with special silver staining H.pylori colonization, HE staining of gastric mucosal inflammation score was significantly lower (P <0.05); A, B, C, D group of gastric tissue homogenate TNF- 28.64 pg / mL ± 12.07 pg / mL, 300.16 pg / mL ± 59.10 pg / mL, 54.96 pg / mL ± 15.02 pg / mL and 503.25 pg / mL ± 1.35 pg / mL respectively; rhLF combined with triple therapy reduced gastric homogenate (P <0.01), and there was significant difference compared with normal saline group, standard triple group and rhLF group.rhLF combined with triple therapy could reduce the expression level of VacA mRNA in gastric homogenate (P <0.01) and Normal saline group, standard triple group and rhLF group were statistically significant.Conclusion: RhLF combined with triple therapy can improve H.pylori eradication rate and reduce gastric mucosal inflammatory response, this effect may be associated with rhLF triple therapy can reduce the release of pro-inflammatory cytokines About rhLF can effectively inhibit the expression of VacA mRNA, suggesting that rhLF has the potential to down-regulate H. pylori toxicity.
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