發(fā)布時間：2018-02-26 08:52

  本文關(guān)鍵詞： 復(fù)發(fā)性口腔潰瘍 腫瘤壞死因子α 白介素6 血管內(nèi)皮生長因子 平瘡方　出處：《南京中醫(yī)藥大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：觀察平瘡方對口腔潰瘍模型大鼠INF-α、IL-6、VEGF的影響,探討該驗方對ROU的治療機(jī)制。方法:將48只SPF級SD大鼠隨機(jī)分成正常對照組(8只),模型鑒定組(8只),模型組(8只),中藥低劑量組(8只),中藥高劑量組(8只),陽性藥對照組(8只)。除正常對照組外,其余各組大鼠于口腔右側(cè)頰部黏膜下注射0.1ml10%醋酸的方法誘導(dǎo)口腔潰瘍發(fā)生。模型鑒定組于造模后第二天肉眼觀察潰瘍發(fā)生情況,脫頸處死后取潰瘍處口腔黏膜組織行HE染色進(jìn)行病理組織學(xué)觀察,驗證造模方法是否成功。其余5組,對正常對照組大鼠和模型組大鼠用蒸餾水進(jìn)行灌胃,中藥低、高劑量組分別以含生藥0.18g/ml和0.9g/ml濃度的水煎液灌胃,陽性對照組用維生素B2和維生素C水溶液灌胃。連續(xù)給藥2周,期間注意觀察各組大鼠一般情況及口腔黏膜變化情況,測量口腔潰瘍模型大鼠潰瘍大小。2周后,將所有大鼠股動脈取血,脫頸處死后取潰瘍處口腔黏膜組織,正常組取對應(yīng)部位正�？谇火つそM織,進(jìn)行HE染色后鏡下觀察潰瘍處口腔黏膜組織的病理變化,血液標(biāo)本離心后仔細(xì)吸取上層清液利用酶聯(lián)吸附法(ELISA)檢測血清INF-α、IL-6、VEGF含量。結(jié)果:1.一般情況觀察:正常對照組大鼠口腔內(nèi)無口腔潰瘍,黏膜呈粉紅色,無黏膜水腫,無假膜形成,其余各組大鼠于造模后第二天,右側(cè)頰部可見類圓形口腔潰瘍發(fā)生,表面凹陷性缺損,覆有黃色假膜,周圍充血水腫。持續(xù)給藥2周內(nèi),各組口腔潰瘍模型大鼠的潰瘍均有所緩解,充血水腫減輕,至給藥14天時,中藥高劑量組和陽性對照組潰瘍面幾乎愈合。各組大鼠進(jìn)食飲水量無明顯區(qū)別。2.HE染色結(jié)果:正常組口腔黏膜組織上皮層、固有層的結(jié)構(gòu)完整,未見炎性細(xì)胞浸潤。模型鑒定組大鼠口腔黏膜破損,上皮結(jié)構(gòu)被破壞,黏膜上皮細(xì)胞破潰脫落壞死,核碎裂、固縮、深染,可見大量空泡變性,潰瘍處有大量炎性細(xì)胞浸潤,以中性粒細(xì)胞和淋巴細(xì)胞為主,與人復(fù)發(fā)性口腔潰瘍的局部病理改變相似,可以證明成功復(fù)制了口腔潰瘍模型。模型組大鼠口腔黏膜仍有破潰,與鑒定組相似,潰瘍處可見大量炎性細(xì)胞浸潤和空泡變性,中藥高劑量組和陽性對照組黏膜上皮組織趨于完整,炎性細(xì)胞浸潤減少,中藥低劑量組仍可見炎性細(xì)胞浸潤,組織結(jié)構(gòu)不完整。3.體重變化:各組大鼠之間體重?zé)o明顯差異。4.潰瘍面積變化:各治療組口腔潰瘍面積明顯小于模型組(P均0.01),中藥高劑量組與陽性對照組治療后潰瘍面積無統(tǒng)計學(xué)差異。5.ELISA結(jié)果:模型鑒定組與模型組血清TNF-α含量明顯升高,與正常組有顯著差異(P0.01),模型組TNF-α含量小于模型鑒定組(P0.05),各治療組TNF-α含量較模型組均有所下降,中藥高劑量組和陽性對照組與模型組相比,TNF-α含量有顯著降低(P0.01),這兩組TNF-α含量雖然比正常對照組稍高,但差異不明顯(P0.05),中藥高劑量組與陽性對照組TNF-α含量差異無統(tǒng)計學(xué)意義(P0.05)。模型鑒定組與模型組血清IL-6含量較正常對照組升高,有顯著差異(P0.01),模型組相比模型鑒定組IL-6水平有所下降,差異有統(tǒng)計學(xué)意義(P0.05),中藥低劑量組IL-6含量較模型組有所減少,差異無統(tǒng)計學(xué)意義(P＞0.05),中藥高劑量組和陽性對照組IL-6含量較模型組也有所下降,差異有統(tǒng)計學(xué)意義(P0.05),中藥高劑量組與陽性對照組IL-6含量差異無統(tǒng)計學(xué)意義(P＞.05),這兩組的IL-6含量接近正常對照組水平,與正常對照組比較差異無統(tǒng)計學(xué)意義(P0.05)。模型組及模型鑒定組大鼠血清VEGF含量較正常對照組顯著升高(P0.01),差異顯著,中藥低劑量組、中藥高劑量組及陽性對照組大鼠血清VEGF含量較模型組有所減少,中藥低劑量組較模型組數(shù)值有所下降,但差異無統(tǒng)計學(xué)意義(P＞0.05),中藥高劑量組與陽性對照組VEGF含量下降顯著(P0.05),中藥高劑量組與陽性對照組大鼠血清VEGF含量差異無統(tǒng)計學(xué)意義(P＞0.05)。結(jié)論:平瘡方能有效減少TNF-α、IL-6的含量,調(diào)控VEGF的表達(dá),達(dá)到抑制炎癥反應(yīng),促進(jìn)口腔潰瘍愈合的作用。
[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.

【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5;R-332

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