黏蛋白MUC2的糖基单一化对Escherichia侵袭力及代谢功能影响及其参与溃疡性结肠炎发病机制研究

Glycosylation of MUC2 on the invasions and metabolic functions of Escherichia and its involvement in the pathogenesis of ulcerative colitis

  • 摘要:
    目的 探讨黏蛋白MUC2的糖基单一化对Escherichia侵袭力及代谢功能的影响及其参与溃疡性结肠炎发病的机制。
    方法 选取溃疡性结肠炎(UC)75例患者纳入研究组,同期选取健康体检者75例纳入对照组。免疫组化检测正常黏膜组织及UC组织中MUC2蛋白水平,采用酶联免疫试剂盒(ELISA)检测C反应蛋白酶(CRP)、粪便钙卫蛋白(FC)水平,全自动生化分析仪检测降钙素原(PCT)水平。Escherichia处理HT29细胞后,分为A组(细胞+MUC2的多聚糖)、B组(细胞+单糖)、C组(细胞+磷酸盐缓冲液)。观察Escherichia生物膜,检测Escherichia黏附力及侵袭能力。聚合酶链反应(PCR)检测LasRpvdApcrVGUS水平。
    结果 正常黏膜组织及UC组织中MUC2蛋白表达水平比较,差异有统计学意义(t=13.770, P < 0.001)。轻度UC患者组织MUC2蛋白水平为(0.63±0.10), 中度UC患者为(0.55±0.09), 重度UC患者为(0.50±0.05), 差异有统计学意义(F=14.880, P < 0.001)。与对照组相比,研究组患者CRP、FC、PCT水平均升高,差异有统计学意义(P < 0.05);与轻度相比,中度患者CRP、FC、PCT水平升高,与中度患者相比,重度患者CRP、FC、PCT水平升高,差异有统计学意义(P < 0.05)。在UC中, MUC2与CRP、FC、PCT水平均呈负相关。A组、B组、C组Escherichia黏附率分别为(15.30±2.55)%、(63.25±8.08)%及(48.61±6.67)%, 差异有统计学意义(F=93.480, P < 0.001)。A组、B组、C组Escherichia侵袭率分别为(24.80±5.11)%、(70.21±10.60)%及(53.34±8.06)%, 差异有统计学意义(F=46.620, P < 0.001)。与A组相比, B组LasRpvdApcrVGUS基因水平升高,与B组相比, C组LasRpvdApcrVGUS基因水平降低,差异有统计学意义(P < 0.05)。
    结论 UC中MUC2表达降低,并随疾病程度的加重而降低,其与CRP、FC、PCT表达呈负相关。Escherichia与MUC2的糖基单一化培养后,可增加其侵袭及黏附能力,以及提高其毒性作用。

     

    Abstract:
    Objective To investigate the effect of glycosylation of MUC2 on the invasions and metabolic functions of Escherichia and its involvement in the pathogenesis of ulcerative colitis.
    Methods A total of 75 patients with ulcerative colitis (UC) were included in the study group, and 75 healthy subjects were included in control group. Immunohistochemistry was used to detect MUC2 protein levels in normal mucosal tissues and UC tissues. C-reactive protease (CRP) and fecal calprotectin (FC) were detected by enzyme-linked immunosorbent assay (ELISA), and procalcitonin (PCT) was detected by automatic biochemical analyzer. HT29 cells were treated with Escherichia, and then divided into group A (cell+MUC2 polysaccharide), group B (cell+monosaccharide) and group C (cell+phosphate buffer). Escherichia biofilm was observed, and the adhesion and invasion ability of Escherichia were detected by AGAR plate. LasR, pvdA, pcrV and GUS levels were detected by polymerase chain reaction (PCR).
    Results The expression level of MUC2 protein in normal mucosa tissues and UC tissues was statistically significant (t=13.770, P < 0.001). The MUC2 protein level was (0.63±0.10) in patients with mild UC, (0.55±0.09) in patients with moderate UC, and (0.50±0.05) in patients with severe UC, and the difference was statistically significant (F=14.880, P < 0.001). Compared with the control group, the levels of CRP, FC and PCT in the study group were statistically increased (P < 0.05). Compared with mild patients, the levels of CRP, FC and PCT in moderate patients were significantly increased, while those in severe patients were significantly increased (P < 0.05); in UC disease, MUC2 was negatively correlated with CRP, FC, and PCT levels. The adhesion rates of Escherichia in group A, group B and group C were (15.30±2.55)%, (63.25±8.08)% and (48.61±6.67)%, respectively (F=93.480, P < 0.001). The invasion rates of Escherichia in groups A, B, and C were (24.80±5.11)%, (70.21±10.60)% and (53.34±8.06)%, respectively, and the differences were statistically significant (F=46.620, P < 0.001). Compared with group A, the levels of LasR, pvdA, pcrV and GUS genes in group B were significantly increased, and the levels of LasR, pvdA, pcrV and GUS genes in group C were significantly decreased compared with group B (P < 0.05).
    Conclusion The expression of MUC2 in UC is decreased and decreased with the severity of the disease, which is negatively correlated with the expression of CRP, FC and PCT. The monosaccharide culture of Escherichia and MUC2 can increase its invasion and adhesion ability, and enhance its toxicity.

     

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