吉林大学学报(医学版) ›› 2022, Vol. 48 ›› Issue (5): 1131-1138.doi: 10.13481/j.1671-587X.20220505

• 基础研究 • 上一篇    

梅花草提取物对幽门螺杆菌小鼠胃黏膜定植的影响及其机制

金昭1,王儒帅1,王朝阳2,何占彪3,田复明3,王睿君3()   

  1. 1.内蒙古医科大学附属医院影像诊断科,内蒙古 呼和浩特 010050
    2.内蒙古医科大学附属医院胃肠外科,内蒙古 呼和浩特 010050
    3.内蒙古医科大学附属医院神经外科 内蒙古自治区神经系统疾病临床医学研究中心,内蒙古 呼和浩特 010050
  • 收稿日期:2022-01-13 出版日期:2022-09-28 发布日期:2022-11-15
  • 通讯作者: 王睿君 E-mail:602507851@qq.com
  • 作者简介:金 昭(1978-),男,内蒙古自治区赤峰市人,副主任医师,医学硕士,主要从事胃肠消化道常见病和多发病影像诊断方面的研究。
  • 基金资助:
    国家自然科学基金项目(82060906);内蒙古自治区科技厅自然科学基金项目(2020MS08146);内蒙古自治区卫健委医疗卫生科技计划项目(202201307);内蒙古医科大学重点项目(YKD2021ZD002)

Effect of Parnassia palustris Linn extracts on Helicobacter pylori colonization in gastric mucosa and its mechanism

Zhao JIN1,Rushuai WANG1,Zhaoyang WANG2,zhanbiao HE3,Fuming TIAN3,Ruijun WANG3()   

  1. 1.Department of Imaging Diagnosis,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China
    2.Department of Gastroenterology Surgery,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China
    3.Clinical Medical Research Center for Neurological Disease,Department of Neurosurgery,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China
  • Received:2022-01-13 Online:2022-09-28 Published:2022-11-15
  • Contact: Ruijun WANG E-mail:602507851@qq.com

摘要:

目的 探讨梅花草提取物对幽门螺杆菌(Hp)根除和胃黏膜的影响,阐明其在体内外抑菌消炎的机制。 方法 6周龄雄性昆明小鼠随机分为空白对照组、生理盐水组、Hp组、三联组、四联组、梅花草组、三联+梅花草组和四联+梅花草组。琼脂稀释法检测梅花草对Hp SS1标准株的最小抑菌浓度(MIC),实时荧光定量PCR(RT-qPCR)法检测Hp定植黏附相关基因 mRNA表达水平。动物实验评估不同治疗方式对Hp的根除率,酶联免疫吸附测定(ELISA)法检测各组小鼠血清中白细胞介素2(IL-2)、白细胞介素8(IL-8)和肿瘤坏死因子α(TNF-α)水平,快速尿素酶试验(RUT)检测各组小鼠胃组织中Hp感染率,HE染色和透射电镜观察小鼠胃组织病理形态表现和超微结构。 结果 梅花草的MIC为10 g·L-1。RT-qPCR法检测,与Hp组比较,梅花草组Hp定植黏膜相关基因Bab A、Cag A和Vac A mRNA表达水平均降低(P<0.05或P<0.01)。免疫组织化学检测,Hp组小鼠胃黏膜部分糜烂和溃疡,出血和充血明显,Hp感染阳性;梅花草组小鼠胃黏膜出血和溃疡明显改善,胃黏膜Hp感染呈阴性。RUT实验检测,与Hp组比较,梅花草组小鼠Hp感染率降低(P<0.05),三联组和四联组小鼠Hp感染率降低(P<0.05);与梅花草组比较,三联组和四联组小鼠Hp根除率差异无统计学意义(P>0.05);与三联或四联组比较,三联+梅花草组和四联+梅花草组小鼠Hp根除率升高(P<0.01)。三联+梅花草组和四联+梅花草组小鼠胃黏膜组织得到修复,胃黏膜上皮细胞的线粒体亚细胞器和微绒毛结构完整。与Hp组比较,三联组、四联组、梅花草组、三联+梅花草组和四联+梅花草组小鼠血清中IL-2水平均升高(P<0.05),IL-8和TNF-α 水平均降低(P<0.05或P<0.01)。 结论 梅花草可以有效根除Hp和保护胃黏膜,其作用机制与降低Hp感染后IL-2、IL-8和TNF-α水平及保护胃黏膜亚细胞结构完整性有关。

关键词: 梅花草, 幽门螺杆菌, 定植, 根除率, 透射电镜, 炎症因子

Abstract:

Objective To explore the effects of Parnassia palustris Linn extracts on Helicobacter pylori (Hp) eradication and gastric mucosa, and to clarify its antibacterial and anti-inflammatory mechanisms in vitro and in vivo. Methods The six-week old male Kunming mice were randomly divided into blank control group, normal saline group, Hp group, triple group, quadruple group, Parnassia palustris Linn group, triple + Parnassia palustris Linn group, and quadruple + Parnassia palustris Linn group. The minimal inhibitory concentration (MIC) of Parnassia palustris Linn extracts on Hp SS1 standard strains in vitro and the expression levels of Hp colonization- and adhesion-related gene mRNA were detected by real-time fluorescence quantitative (RT-qPCR) method. Animal experiments were used to evaluate the eradication rate of Hp by different methods, and enzyme linked immunosorbent assay(ELISA) method was used to detect the serum levels of inflammatory factors such as interleukin-2 (IL-2), interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-α) of the mice in various groups. Rapid urease test (RUT) was used to detect the infection rate of Hp in stomach tissue, HE staining and transmission electron microscope were used to detect the pathomorphology and ultrastructures of stomach tissue. Results The MIC of Parnassia palustris Linn was 10 g·L-1.The RT-qPCR results showed that compared with Hp group,the expression levels of Bab A, Cag A and Vac A mRNA in Parnassia palustris Linn group were decreased (P<0.05 or P<0.01).The immunohistochemistry results showed that the part of gastric mucosa had erosion and ulcers,obvious bleeding and conggestion,and positive Hp infection of the mice in Hp group; the symptoms of gastric mucosal bleeding and ulcers in Parnassia palustris Linn group were improved, and Hp infection was mostly negative.The Rut results showed that compared with Hp group,the infection rate of Hp of the mice in Parnassia palustris Linn group was decreased(P<0.05),and the infection rates of the mice in triple group and quadruple group were decreased(P<0.05);compared with Parnassia palustris Linn group,the eradication rates of Hp of the mice in triple group and quadruple group had no significant differences(P>0.05); compared with triple group or quadruple group, the eradication rates of Hp of the mice in triple+Parnassia palustris Linn group and quadruple+Parnassia palustric Linn group were increased(P<0.01). The gastric mucosa was apparently repaired, and the structures of mitochondria and microvilli of gastric mucosal epithelial cells were intact of the mice in triple+Parnassia palustris Linn group and quadruple+Parnassia palustris Linn group.Compared with Hp group,the levels of IL-2 in serum of the mice in triple, quadruple,Parnassia palustris Linn, triple+Parnassia palustris Linn and quadruple+Parnassia palustris Linn groups were increased(P<0.05), and the levels of serum IL-8 and TNF-α were decreased(P<0.05 or P<0.01). Conclusion Parnassia palustris Linn extracts can effectively eradicate Hp and protect gastric mucosa, and its mechanism is related to decreasing the levels of IL-2,IL-8 and TNF-α and protecting the integrity of structures of gastric mucosa subcellular organelle after Hp infection.

Key words: Parnassia palustris Linn, Helicobacter pylori, Colonization, Eradication rate, Transmission electron microscope, Inflammatory factors

中图分类号: 

  • R856.5