小毛莨内酯对脊柱结核兔模型病灶组织中IL-6，TNF-a和TGF-β的影响

发布时间：2018-04-02 14:00

本文选题：脊柱结核　切入点：白介素-6　出处：《南华大学》2014年硕士论文

【摘要】：结核病为一慢性可治愈但死亡率最高之传染病，特别是脊柱结核的发病率与日俱增。近年来耐药及多重耐药菌株(multiple drugs resistant-bacilustuberculosis,MDR-TB)的出现,更是对现有的抗结核药物提出了严重挑战，寻找新的抗结核药，尤其是抗结核中药已成为研究方向。有研究显示IL-6、TNF-a和TGF-β在结核病变转归过程中起到重要作用，发现猫爪草与抗结核药物结合可以提高疗效，缩短疗程，且没有产生毒副作用，还能提高患者的免疫力，但目前其机制不明。目的：观察小毛莨内酯对脊柱结核兔病灶组织中白细胞介素-6（IL-6），肿瘤坏死因子（TNF-a），转化生长因子-β（TGF-β）的表达水平的影响，探讨其治疗脊柱结核的作用机制。方法：将未成年兔腰4椎体钻孔植入结核标准菌株H37Rv，建立兔脊柱结核模型。将20只新西兰未成年兔随机分成4组，每组5只。A组，脊柱结核造模组，不予以任何干预治疗；B组，小毛莨内酯治疗组，即脊柱结核造模后给予小毛莨内酯灌胃治疗；C组：常规抗痨药治疗组，脊柱结核造模后予以常规抗痨药灌胃治疗；D组：常规抗痨药联合小毛莨内酯治疗组，脊柱结核造模后分别予以常规抗痨药及小毛莨内酯灌胃治疗；8周后，取病灶组织，进行HE染色和肉眼组织学观察，实时荧光定量PCR反应法及免疫印迹法分别检测小毛莨内酯及常规三联抗痨药治疗前后脊柱结核兔病灶组织中IL-6、TNF-α、TGF-β的mRNA和蛋白表达水平的变化。结果应用SPSS16.0统计软件，进行方差分析（on wayANOVA），分析各组间差异， P0.05表示差异性显著。结果：新西兰白兔腰椎进行H37Rv结核杆菌感染后，结核病灶组织HE染色显示组织间有脓细胞、类上皮样细胞或坏死灶等形成，经小毛莨内酯、常规抗捞药以及联合用药治疗8周后，结核病灶区脓细胞、类上皮样细胞数量显著减少；实时荧光定量PCR反应监测腰椎结核病灶组织中脊柱结核造模组的IL-6、TNF-α、TGF-βmRNA相对表达量与小毛莨内酯治疗组、常规抗捞药治疗组及常规抗捞药联合小毛莨内酯治疗组相比较有显著差异（P0.05），脊柱结核兔模型经过治疗后病灶组织中IL-6、TNF-α、TNF-αmRNA表达明显降低。常规抗捞药联合小毛莨内酯治疗组的IL-6、TNF-α、TGF-βmRNA相对表达量与小毛莨内酯治疗组及常规抗捞药治疗组相比较有显著差异（P0.05）,且常规抗捞药联合小毛莨内酯治疗组中IL-6、TNF-α、TGF-βmRNA的相对表达量最低，提示采用常规抗捞药联合小毛莨内酯治疗后，更能降低组织中IL-6、TNF-α、TGF-βmRNA的表达水平；免疫印迹法检测各组椎旁软组织中IL-6、TNF-α、TGF-β的蛋白表达水平与荧光定量PCR检测结果类似。结论： 1.小毛莨内酯可以有效降低脊柱结核兔模型病灶组织中炎症相关因子IL-6，，TNF-a的表达水平，减轻炎性反应，降低TGF-β的表达水平，抑制病灶中肉芽组织形成。 2．小毛莨内酯对常规抗结核药治疗结核病有协同作用。
[Abstract]:Tuberculosis is a chronic curable infectious disease with the highest mortality, especially spinal tuberculosis.Some studies have shown that IL-6, TNF-a and TGF- 尾 play an important role in the process of tuberculosis progression.But at present its mechanism is not clear.Twenty New Zealand minor rabbits were randomly divided into 4 groups: group A (n = 5), group B (n = 5) with spinal tuberculosis, group B without any intervention, group B (n = 5) treated with rhinolactone, group A (n = 5), group B (n = 5) with spinal tuberculosis.The spinal tuberculosis model was treated with routine antituberculous drugs and rhinolactone respectively for 8 weeks. The lesion tissues were taken for HE staining and gross histological observation.The changes of mRNA and protein expression of IL-6 TNF- 伪 TGF- 尾 were detected by real-time quantitative PCR reaction and Western blotting before and after treatment with rhinolactone and conventional anti-tuberculosis drugs in rabbits with spinal tuberculosis.Results using SPSS16.0 statistical software, ANOVA was used to analyze the differences among the groups, which showed significant difference (P0.05).Results: after H37Rv infection in the lumbar vertebrae of New Zealand white rabbits, HE staining showed that there were pyogenic cells, epithelioid cells or necrotic foci between the tissues.After 8 weeks of treatment, the number of pyogenic cells and epithelioid cells in tuberculosis foci decreased significantly.Real-time fluorescence quantitative PCR reaction was used to detect the relative expression of IL-6 TNF- 伪 TGF- 尾 mRNA in the spinal tuberculosis model group and the treatment group.The expression of IL-6 TNF- 伪 TNF- 伪 mRNA in spinal tuberculosis rabbit model was significantly lower than that in the conventional anti-salvage therapy group and the conventional anti-salvage drug combined with rhinolactone treatment group (P0.05), and the expression of IL-6 TNF- 伪 TNF- 伪 was significantly decreased in the spinal tuberculosis model after treatment.The relative expression of IL-6 TNF- 伪 TGF- 尾 mRNA in the conventional anti-salvage drugs combined with Ranunculus milunculatus group was significantly different from that in the Ranunculus milunculatus treatment group and the routine anti-salvage drug treatment group, and the relative expression of IL-6TNF- 伪 TGF- 尾 mRNA was the lowest in the conventional anti-salvage drug combined with Ranunculum milunculate treatment group, and the relative expression of IL-6 TNF- 伪 TGF- 尾 mRNA was the lowest in the conventional anti-salvage drug combined with Ranunculum milunculus lactone treatment group.Conclusion:1.Rhinolactone can effectively reduce the expression of IL-6 and TNF-a in the focal tissue of spinal tuberculosis rabbit model, alleviate the inflammatory reaction, reduce the expression of TGF- 尾, and inhibit the granulation tissue formation in the focus.2.Rhinolactone has a synergistic effect on the treatment of tuberculosis with routine antituberculosis drugs.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R529.2

【参考文献】