平疮方对口腔溃疡模型大鼠TNF-α、IL-6和VEGF的影响

发布时间：2018-02-26 08:52

本文关键词： 复发性口腔溃疡肿瘤坏死因子α 白介素6 血管内皮生长因子平疮方　出处：《南京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的：观察平疮方对口腔溃疡模型大鼠INF-α、IL-6、VEGF的影响,探讨该验方对ROU的治疗机制。方法:将48只SPF级SD大鼠随机分成正常对照组(8只),模型鉴定组(8只),模型组(8只),中药低剂量组(8只),中药高剂量组(8只),阳性药对照组(8只)。除正常对照组外,其余各组大鼠于口腔右侧颊部黏膜下注射0.1ml10%醋酸的方法诱导口腔溃疡发生。模型鉴定组于造模后第二天肉眼观察溃疡发生情况,脱颈处死后取溃疡处口腔黏膜组织行HE染色进行病理组织学观察,验证造模方法是否成功。其余5组,对正常对照组大鼠和模型组大鼠用蒸馏水进行灌胃,中药低、高剂量组分别以含生药0.18g/ml和0.9g/ml浓度的水煎液灌胃,阳性对照组用维生素B2和维生素C水溶液灌胃。连续给药2周,期间注意观察各组大鼠一般情况及口腔黏膜变化情况,测量口腔溃疡模型大鼠溃疡大小。2周后,将所有大鼠股动脉取血,脱颈处死后取溃疡处口腔黏膜组织,正常组取对应部位正常口腔黏膜组织,进行HE染色后镜下观察溃疡处口腔黏膜组织的病理变化,血液标本离心后仔细吸取上层清液利用酶联吸附法(ELISA)检测血清INF-α、IL-6、VEGF含量。结果:1.一般情况观察:正常对照组大鼠口腔内无口腔溃疡,黏膜呈粉红色,无黏膜水肿,无假膜形成,其余各组大鼠于造模后第二天,右侧颊部可见类圆形口腔溃疡发生,表面凹陷性缺损,覆有黄色假膜,周围充血水肿。持续给药2周内,各组口腔溃疡模型大鼠的溃疡均有所缓解,充血水肿减轻,至给药14天时,中药高剂量组和阳性对照组溃疡面几乎愈合。各组大鼠进食饮水量无明显区别。2.HE染色结果:正常组口腔黏膜组织上皮层、固有层的结构完整,未见炎性细胞浸润。模型鉴定组大鼠口腔黏膜破损,上皮结构被破坏,黏膜上皮细胞破溃脱落坏死,核碎裂、固缩、深染,可见大量空泡变性,溃疡处有大量炎性细胞浸润,以中性粒细胞和淋巴细胞为主,与人复发性口腔溃疡的局部病理改变相似,可以证明成功复制了口腔溃疡模型。模型组大鼠口腔黏膜仍有破溃,与鉴定组相似,溃疡处可见大量炎性细胞浸润和空泡变性,中药高剂量组和阳性对照组黏膜上皮组织趋于完整,炎性细胞浸润减少,中药低剂量组仍可见炎性细胞浸润,组织结构不完整。3.体重变化:各组大鼠之间体重无明显差异。4.溃疡面积变化:各治疗组口腔溃疡面积明显小于模型组(P均0.01),中药高剂量组与阳性对照组治疗后溃疡面积无统计学差异。5.ELISA结果:模型鉴定组与模型组血清TNF-α含量明显升高,与正常组有显著差异(P0.01),模型组TNF-α含量小于模型鉴定组(P0.05),各治疗组TNF-α含量较模型组均有所下降,中药高剂量组和阳性对照组与模型组相比,TNF-α含量有显著降低(P0.01),这两组TNF-α含量虽然比正常对照组稍高,但差异不明显(P0.05),中药高剂量组与阳性对照组TNF-α含量差异无统计学意义(P0.05)。模型鉴定组与模型组血清IL-6含量较正常对照组升高,有显著差异(P0.01),模型组相比模型鉴定组IL-6水平有所下降,差异有统计学意义(P0.05),中药低剂量组IL-6含量较模型组有所减少,差异无统计学意义(P＞0.05),中药高剂量组和阳性对照组IL-6含量较模型组也有所下降,差异有统计学意义(P0.05),中药高剂量组与阳性对照组IL-6含量差异无统计学意义(P＞.05),这两组的IL-6含量接近正常对照组水平,与正常对照组比较差异无统计学意义(P0.05)。模型组及模型鉴定组大鼠血清VEGF含量较正常对照组显著升高(P0.01),差异显著,中药低剂量组、中药高剂量组及阳性对照组大鼠血清VEGF含量较模型组有所减少,中药低剂量组较模型组数值有所下降,但差异无统计学意义(P＞0.05),中药高剂量组与阳性对照组VEGF含量下降显著(P0.05),中药高剂量组与阳性对照组大鼠血清VEGF含量差异无统计学意义(P＞0.05)。结论:平疮方能有效减少TNF-α、IL-6的含量,调控VEGF的表达,达到抑制炎症反应,促进口腔溃疡愈合的作用。
[Abstract]:Objective: To observe the level of INF- alpha sores, oral ulcer rat model of IL-6, the influence of VEGF, to investigate the treatment mechanism of the prescription of ROU. Methods: 48 SPF SD rats were randomly divided into normal control group (8 rats), model group (8 rats), identification of the model group (8 rats Chinese Medicine), the low dose group (8 rats), high dose of Chinese medicine group (8 rats), positive control group (8 rats). Except the normal control group, oral ulcer induced by other groups of rats in the oral cavity, the right buccal submucosal injection of 0.1ml10% acetate. The model identification group after modeling the second day the naked eye to observe the occurrence of ulcer, were sacrificed after the ulcer of oral mucosal tissue for histopathological observation HE staining, verify the modeling method is successful. The other 5 groups of rats in the normal control group and model group rats with distilled water gavage, traditional Chinese medicine low and high dose group respectively with drug 0.18g/ml and 0.9g/ml concentration in water decoction Liquid gavage, positive control group with vitamin B2 and vitamin C in aqueous solution by gavage for 2 weeks. During the period, to observe the general conditions of rats and changes of oral mucosa, oral ulcer rat model of measuring ulcer size after.2 weeks, all the rats blood from the femoral artery from the ulcer of oral mucosa groups were sacrificed after normal group to take corresponding parts of normal oral mucosa, pathological changes were observed under microscope after HE staining, the ulcer of oral mucosa, blood samples after centrifugation and the supernatant by carefully draw enzyme-linked immunosorbent assay (ELISA) detection of serum alpha INF-, IL-6, VEGF content. Results: 1. in general observation: the normal control group rats without oral oral ulcer, mucous membrane is pink, no mucosal edema, no pseudomembrane formation, the rest of the rats after modeling for second days, the right buccal visible circular oral ulcers, surface depression, cover Yellow surrounding hyperemia and edema. Pseudomembrane, lasting for 2 weeks, each group oral ulcer model rat ulcer were relieved to reduce congestion and edema, administered for 14 days, Chinese medicine high dose group and positive control group surface ulcer healing. Almost the rats eating drinking no difference.2.HE staining results normal group: oral mucosa epithelium, lamina propria of the complete structure, no inflammatory cell infiltration in model rats. Identification of oral mucosa, epithelial structure was destroyed, epithelial cells rupture shedding necrosis, nuclear fragmentation, pyknosis, anachromasis, vacuoles degeneration, ulcers and a large number of inflammatory cells the infiltration of neutrophils and lymphocytes, and local pathological changes similar to recurrent oral ulcer, that successfully replicated oral ulcer models. The oral mucosa of rats in the model group is still a rupture, and identification were similar, the ulcer may be To see a large number of inflammatory cell infiltration and vacuolar degeneration, high dose of Chinese medicine group and positive control group of epithelial tissue integrity, inflammatory cell infiltration, low dose of Chinese medicine group is still visible inflammatory cell infiltration, the organizational structure is not complete.3. weight change: the weight of rats in each group did not change significantly.4.: the treatment of ulcer area oral ulcer area was significantly lower than that of the model group (P 0.01), Chinese medicine high dose group and positive control group after treatment, the ulcer area had no significant difference.5.ELISA identification results: model group and model group of serum TNF- was significantly increased, there was significant difference between normal group and model group (P0.01), TNF- content was smaller than that in the model group identification (P0.05), alpha TNF- in each treatment group compared with that in the model group were decreased, compared with the traditional Chinese medicine high dose group and positive control group and model group, TNF- content was decreased significantly (P0.01), the two group of TNF- alpha although the content is The control group is slightly higher, but the difference was not significant (P0.05), there was no significant difference between Chinese traditional medicine high dose group and positive control group TNF- (P0.05). The content of alpha model identification group and model group of serum IL-6 content increased compared to normal control group, there was significant difference (P0.01), model group compared to the IL-6 level of model group identification decreased, the difference was statistically significant (P0.05), low dose of Chinese medicine group IL-6 compared with that in the model group decreased, the difference was not statistically significant (P > 0.05), Chinese medicine high dose group and positive control group IL-6 compared with that in the model group decreased, the difference was statistically significant (P0.05), no statistically significant difference between Chinese high dose group and positive control group IL-6 content (P >.05), the content of IL-6 of the two groups close to the level of normal control group and normal control group, no significant difference (P0.05). The model group and model identification group rats serum VEGF content compared with the normal control group Significantly increased (P0.01), significant difference, low dosage of Chinese medicine group, Chinese medicine high dose group and positive control group rats serum VEGF content decreased compared with the model group, low dose of Chinese medicine group than the control group values decreased, but the difference was not statistically significant (P > 0.05), Chinese Medicine high dose group and positive control group the content of VEGF decreased significantly (P0.05), high dose of Chinese medicine group and positive control group rats serum VEGF content difference was not statistically significant (P > 0.05). Conclusion: flat wound side can effectively reduce the content of IL-6, TNF- alpha, regulating the expression of VEGF, inhibit inflammatory reaction, promote oral ulcer healing.

【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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