白芨多糖对胃溃疡大鼠的干预作用及其机制研究

目的研究白芨多糖对胃溃疡(GU)模型大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及病灶组织的核因子-κB(NF-κB)p65基因及其蛋白表达影响。方法用乙酸直接烧灼法复制GU大鼠模型。按随机数字表法将大鼠分为6组:正常组、模型组,阳性对照组(0.3 g·kg^-1·d^-1雷尼替丁灌胃)和大、中、小3个剂量实验组(50.0,25.0,12.5 mg·kg^-1·d^-1白芨多糖灌胃),每组10只;正常组和模型组大鼠给予蒸馏水灌胃,各组均1次/天,连续15 d。用酶联免疫吸附法检测大鼠血清中TNF-α、IL-1β和IL-6含量,以实时荧光定量PCR法和蛋白免疫印迹法检测组织中NF-κB基因和蛋白表达情况。结果正常组、模型组、阳性对照组和大、中、小3个剂量实验组的血清TNF-α含量分别为(571.13±11.85),(759.25±21.11),(636.63±3.70),(611.75±12.46),(741.63±11.99)和(756.88±8.34)pg·mL^-1;这6组的IL-1β的含量分别为(400.50±30.39),(532.13±11.76),(458.88±9.48),(422.00±21.50),(508.63±15.68)和(523.38±10.18)pg·mL^-1;这6组的IL-6的含量分别为(295.63±11.15),(429.00±11.56),(340.63±9.68),(317.88±10.80),(377.50±14.15)和(417.75±9.36)pg·mL^-1;这6组的NF-κB p65基因表达量(2-ΔΔCt值)分别为1.00±0.00,1.37±0.02,1.29±0.02,1.23±0.03,1.33±0.02和1.35±0.03;这6组的NF-κB p65蛋白表达量(灰度值)分别为0.60±0.11,1.24±0.36,0.79±0.14,0.63±0.09,0.87±0.15和1.16±0.25。上述指标:模型组与正常组比较,差异均有统计学意义(均P<0.01),大、中2个剂量实验组与模型组比较,差异均有统计学意义(P<0.01,P<0.05)。结论白芨多糖可通过抑制促炎因子TNF-α、IL-1β和IL-6异常分泌,并下调NF-κB p65基因及其蛋白表达,从而发挥保护胃黏膜的作用。

Study on the intervention effect of Bletilla striata polysaccharide on gastric ulcer rats and its mechanism

Objective To study the effect of Bletilla striata polysaccharide(BSP) on serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6) and nuclear factor-kappa B(NF-κB) p65 gene and its protein expression in gastric ulcer(GU)rats. Methods The GU rat model was induced by acetic acid direct cauterization. According to the method of random number table,the experimental rats were divided into 6 groups: normal group,model group, positive control group(0.3 g·kg^-1·d^-1 ranitidine) and large,middle and small dose experimental groups( 50. 0,25. 0,12. 5 mg·kg^-1·d^-1 BSP) with 10 rats in each group;the rats in the normal group and the model group were given distilled water,once a day for 15 days. The contents of TNF-α,IL-1β and IL-6 in serum of rats were detected by enzyme linked immunosorbent assay. The expression of NF-κB p65 gene and protein in gastric ulcer lesions were detected by real time fluorescence quantitative PCR and Western blot,respectively. Results The levels of TNF-α in serum of normal group,model group,positive control group and large,medium and small dose experimental groups were( 571. 13 ± 11. 85),( 759. 25 ± 21. 11),( 636. 63 ± 3. 70),( 611. 75 ± 12. 46),( 741. 63 ± 11. 99) and( 756. 88 ± 8. 34) pg·mL^-1,respectively;the levels of IL-1β in the six groups were( 400. 50 ± 30. 39),( 532. 13 ± 11. 76),( 458. 88 ± 9. 48),( 422. 00 ± 21. 50),( 508. 63 ± 15. 68) and( 523. 38 ± 10. 18) pg · mL^-1,respectively;the levels of IL-6 in the six groups were( 295. 63 ± 11. 15),( 429. 00 ± 11. 56),( 340. 63 ± 9. 68),( 317. 88 ± 10. 80),( 377. 50 ± 14. 15) and( 417. 75 ± 9. 36) pg·mL^-1,respectively;the expression( 2-ΔΔCt value) of NF-κB p65 gene in the six groups were 1. 00 ± 0. 00,1. 37 ± 0. 02,1. 29 ± 0. 02,1. 23 ± 0. 03,1. 33 ± 0. 02 and 1. 35 ± 0. 03,respectively;the expression( grey value) of NF-κB p65 protein in the six groups were 0. 60 ± 0. 11,1. 24 ± 0. 36,0. 79 ± 0. 14,0. 63 ± 0. 09,0. 87 ± 0. 15 and 1. 16 ± 0. 25,respectively. Comparison between model group and normal group,the difference of the factors were significantly( all P < 0. 01). Comparison between large,middle dose experimental groups and model group,the difference of the factors were significantly( P < 0. 01,P < 0. 05). Conclusion BSP can protect gastric mucosa by inhibiting the abnormal secretion of pro-inflammatory factors TNF-α,IL-1β and IL-6,and down-regulating the expression of NF-κB p65 gene and its protein.