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  本文關(guān)鍵詞：5味山姜屬中藥對胃潰瘍寒證大鼠溫胃散寒藥效及作用機制研究　出處：《廣州中醫(yī)藥大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 山姜屬中藥 胃潰瘍寒證 溫胃散寒 藥效研究 作用機理

【摘要】：目的：篩選冰乙酸灌胃制作胃潰瘍寒證大鼠模型的最佳造模方法,觀察5味山姜屬中藥對冰乙酸灌胃型胃潰瘍寒證的行為體征、胃液量、胃酸、胃蛋白酶、胃組織病理變化,闡釋此5味山姜屬中藥的溫胃散寒功效本質(zhì)；測定胃組織PGE2、EGF、TGF-a,血清N0、IL-8、TNF-α、NT,血漿ET,腦組織AchE、5-HT、NE的含量,探索5味山姜屬中藥治療胃潰瘍寒證的作用機理。方法：設(shè)置5%、10%、15%、20%、25%5個冰乙酸組,加4℃知母水煎液制作胃潰瘍寒證大鼠模型,比較潰瘍面積、潰瘍指數(shù)、致死性、病理組織變化以及自然修復(fù)情況,確定出最佳的冰乙酸灌胃濃度。最佳灌胃濃度確定后,設(shè)置空白對照組、病證模型對照組、2個陽性對照組、10個受試藥物組,每組10只。除空白對照組外,其余各組采用最終確定的冰乙酸濃度加4℃知母水煎液造模,造模成功后,空白對照組、模型對照組灌服蒸餾水,其余各組分別灌服各受試藥物。觀察胃潰瘍寒證大鼠行為表征的變化(進(jìn)食量、體重、毛色、大便、精神狀態(tài))；測定胃液量、胃酸、胃蛋白酶活力；HE染色觀察胃組織病理變化情況；采用酶聯(lián)免疫法測定PGE2、EGF、TGF-α、IL-8、 TNF-β、NT、血漿ET、5-HT、NE的含量；硝酸還原法測定N0含量；蛋白定量法測定AchE含量。采用SPSS 19.0進(jìn)行統(tǒng)計學(xué)處理,以P0.05為有統(tǒng)計學(xué)意義。結(jié)果：一、寒涼藥加冰乙酸灌胃建立胃潰瘍寒證大鼠模型①15%冰乙酸濃度效果最好,形成的潰瘍大,未見致死性,且自然恢復(fù)的慢。②冰乙酸灌胃復(fù)制胃潰瘍寒證大鼠模型的最佳造模方法為：給予大鼠灌服4℃冰知母水煎液和15%冰乙酸溶液,各1次/d,8：00灌服冰知母,18：00灌服冰乙酸,連續(xù)4 d。室內(nèi)環(huán)境設(shè)定溫度16℃,濕度60%。二、5味山姜屬中藥對胃潰瘍寒證大鼠溫胃散寒藥效研究(一)對胃潰瘍寒證大鼠行為體征的影響：①對胃潰瘍寒證大鼠體重的影響：與模型組比較,10個受試藥物組大鼠體重均增加(P0.05或P0.01)。②對胃潰瘍寒證大鼠進(jìn)食量的影響：與模型組比較,10個受試藥物組大鼠進(jìn)食量均增加,增加率升高明顯。③對胃潰瘍寒證大鼠毛色評分值的影響：與模型組比較,10個受試藥物組,除大高良姜低劑量組外,各組大鼠毛色評分值均降低(P0.05或P0.01)。④對胃潰瘍寒證大鼠大便、精神狀態(tài)的影響：受試藥物大鼠大便量均減少,較干爽,成形；精神狀態(tài)有所恢復(fù),行動較靈活。(二)對胃液量、胃酸、胃蛋白酶活力的影響與模型組比較,大高良姜高劑量、高良姜高劑量、高良姜低劑量、草豆蔻高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量、益智高劑量能減少胃液的分泌量(P0.05或P0.01)；大高良姜高劑量、高良姜高劑量、高良姜低劑量、草豆蔻高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量、益智高劑量、益智低劑量組pH升高分泌和胃蛋白酶活力降低(P0.05或P0.01)。(三)對胃組織病理組織變化的影響與模型組比較,大高良姜高劑量組炎細(xì)胞浸潤減少,粘膜表面滲出物減少,肉芽組織增多,粘膜表層出現(xiàn)新生上皮細(xì)胞排列；高良姜高劑量組表面壞死組織減少,肌層、粘膜下層粘膜肌層結(jié)構(gòu)清晰,肉芽組織增多,粘膜表面上皮組織整齊排列；草豆蔻高劑量組炎細(xì)胞浸潤減少,表面滲出物減少,肉芽組織增多,表面開始出現(xiàn)上皮修復(fù)；紅豆蔻高劑量組炎細(xì)胞浸潤減少,肉芽組織均勻分布,粘膜表面滲出物消失,新生上皮組織逐漸覆蓋粘膜表面；益智高劑量組炎細(xì)胞浸潤減少,部分粘膜表面出現(xiàn)上皮修復(fù)；大高良姜低劑量組粘膜表面滲出物減少,肉芽組織增多,粘膜表面有新生腺體覆蓋；高良姜低劑量組粘膜表面滲出物減少,粘膜層覆蓋新生上皮組織；草豆蔻低劑量組肌層、粘膜下層結(jié)構(gòu)清晰,斷裂的粘膜肌層開始修復(fù),粘膜表面覆蓋上皮組織；紅豆蔻低劑量組粘膜表面滲出物減少,炎細(xì)胞浸潤減少,肉芽組織增多,粘膜肌層開始修復(fù)；益智低劑量組肌層結(jié)構(gòu)完整,粘膜下層布滿肉芽組織。三、5味山姜屬中藥對胃潰瘍寒證大鼠溫胃散寒作用機制研究(一)對胃潰瘍寒證大鼠胃組織PGE2含量變化的影響與模型組比較,高良姜高劑量、草豆蔻高劑量組大鼠胃組織PGE2含量明顯升高,結(jié)果有顯著性差異(P0.01)；大高良姜高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量組大鼠胃組織PGE2含量明顯升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜低劑量、高良姜低劑量、益智高劑量、益智低劑量組大鼠胃組織PGE2含量變化不明顯(P0.05)。(二)對胃潰瘍寒證大鼠胃組織EGF含量的影響與模型對照組比較,大高良姜高劑量、草豆蔻高劑量、益智高劑量組大鼠胃組織EGF含量明顯升高,結(jié)果有顯著性差異(P0.01)；高良姜高劑量、草豆蔻低劑量、紅豆蔻高劑量組胃組織EGF含量升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜低劑量、紅豆蔻低劑量、益智低劑量組胃組織EGF變化無差異(P0.05)。(三)對胃潰瘍寒證大鼠胃組織TGF-a含量的影響與模型組比較,大高良姜高劑量、大高良姜低劑量、高良姜高劑量組、草豆蔻高劑量、紅豆蔻高劑量、益智高劑量組大鼠胃組織TGF-α含量明顯升高,結(jié)果有顯著性差異(P0.01)；高良姜低劑量、草豆蔻低劑量、紅豆蔻低劑量、益智低劑量組大鼠胃組織TGF-α含量變化不明顯(P0.05)。(四)對胃潰瘍寒證大鼠血清N0含量的影響與模型組比較,紅豆蔻高劑量以及附子理中丸、西咪替丁組大鼠血清N0含量明顯升高,結(jié)果有顯著性差異(P0.01)；大高良姜高劑量、高良姜高劑量、草豆蔻高劑量、紅豆蔻低劑量、益智高劑量組大鼠胃組織N0含量升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜低劑量組、高良姜低劑量、草豆蔻低劑量、益智低劑量組變化不明顯(P0.05)。(五)對胃潰瘍寒證大鼠血清IL-8含量的影響與模型組比較,草豆蔻高劑量、紅豆蔻高劑量、益智高劑量組以及附子理中丸、西咪替丁組大鼠胃組織IL-8含量明顯降低,結(jié)果有顯著性差異(P0.01)；大高良姜高劑量、大高良姜低劑量、高良姜高劑量、高良姜低劑量、草豆蔻低劑量、紅豆蔻低劑量、益智低劑量組大鼠胃組織IL-8含量升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)。(六)對胃潰瘍寒證大鼠血清TNF-α含量的影響與模型組比較,大高良姜高劑量、大高良姜低劑量、高良姜高劑量、高良姜低劑量、草豆蔻高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量、益智高劑量、益智低劑量組大鼠血清TNF-α含量變化均無差異(P0.05)。(七)對胃潰瘍寒證大鼠血清NT含量的影響與模型組比較,大高良姜高劑量、大高良姜低劑量、高良姜高劑量、高良姜低劑量、草豆蔻高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量、益智高劑量組大鼠血清NT含量明顯升高(P0.01)。益智低劑量組大鼠血清NT含量升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)。(八)對胃潰瘍寒證大鼠血漿ET含量的影響與模型組比較,高良姜高劑量、草豆蔻高劑量、草豆蔻低劑量、紅豆蔻高劑量、紅豆蔻低劑量組大鼠血漿ET含量明顯降低(P0.01)。大高良姜高劑量、大高良姜低劑量、高良姜低劑量、益智高劑量、益智低劑量組大鼠血漿ET含量降低,結(jié)果有統(tǒng)計學(xué)意義(P0.05)。(九)對胃潰瘍寒證大鼠腦組織AchE含量的影響與模型對照組比較,高良姜高劑量、草豆蔻高劑量、紅豆蔻高劑量組大鼠腦組織AchE含量明顯升高(P0.01)；大高良姜高劑量、高良姜低劑量、紅豆蔻高劑量組大鼠腦組織AchE含量升高,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜低劑量、草豆蔻低劑量、紅豆蔻低劑量、益智低劑量組大鼠腦組織AchE含量變化無差異(P0.05)。(十)對胃潰瘍寒證大鼠腦組織5-HT含量的影響與模型對照組比較,大高良姜高劑量、高良姜高劑量、草豆蔻高劑量、紅豆蔻高劑量、紅豆蔻低劑量、益智高劑量、益智低劑量組大鼠腦組織5-HT含量明顯降低(P0.01)。草豆蔻低劑量組大鼠腦組織5-HT含量降低,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜低劑量、高良姜低劑量組大鼠腦組織5-HT無明顯變化(P0.05)。(十一)對胃潰瘍寒證大鼠腦組織NE含量的影響與模型對照組比較,草豆蔻高劑量、紅豆蔻高劑量、益智高劑量組大鼠腦組織NE含量明顯降低(P0.01)。紅豆蔻低劑量組大鼠腦組織NE含量降低,結(jié)果有統(tǒng)計學(xué)意義(P0.05)；大高良姜高劑量、大高良姜低劑量、高良姜高劑量、高良姜低劑量、草豆蔻低劑量、益智低劑量組大鼠腦組織NE無明顯變化(P0.05)。結(jié)論：一、冰乙酸灌胃型大鼠胃潰瘍寒證大鼠模型的最佳冰乙酸濃度為15%。二、冰乙酸灌胃型胃潰瘍寒證與PGE2、EGF、TGF-α、NT、NO、AchE含量降低有關(guān)；與 IL-8、TNF-α、ET、5-HT、NE升高含量有關(guān)。三、5味山姜屬中藥治療冰乙酸型胃潰瘍寒證的作用機理可能與減少胃液分泌,降低胃酸酸度、胃蛋白酶活力有關(guān)；與升高PGE2、EGF、TGF-α、NT、NO、AchE含量有關(guān)；與降低IL-8、血漿ET、5-HT、NE含量有關(guān)。四、存在親緣關(guān)系的大高良姜、高良姜、草豆蔻、紅豆蔻、益智這5味中藥,在溫胃散寒功效上存在相似的藥理效應(yīng)。
[Abstract]:Objective: the best method for screening acetic acid made mould made by filling the stomach ulcer with cold syndrome rat model, observation of 5 kinds of Alpinia herbs on acetic acid gastric ulcer gastric perfusion cold syndrome behavioral signs, volume of gastric juice, gastric acid, pepsin, pathological changes of gastric tissue, warm the stomach illustrates this 5 kinds of traditional Chinese medicine for dispelling cold Alpinia effect of nature; the gastric tissue was measured by PGE2, EGF, TGF-a, serum N0, IL-8, TNF- alpha, NT, ET in plasma, brain AchE, 5-HT, NE content, explore the mechanism of 5 kinds of traditional Chinese medicine in the treatment of gastric ulcer in Alpinia cold syndrome. Methods: 5%, 10%, 15%, 20%, 25%5 ice acetic acid group, plus 4 degrees Zhimu Decoction made cold syndrome rat model of gastric ulcer, ulcer area and ulcer index, comparison, lethality, histopathological changes and natural restoration, determined the optimal concentration of acetic acid by gavage. The best concentration was determined after intragastric administration, blank control group, disease model group. 2 positive controls Group, 10 subjects in drug group, 10 rats in each group. Except control group, other groups were finally determined the concentration of acetic acid plus 4 DEG C Zhimu Decoction model, after the success of the model, the blank control group, model control group with distilled water, the other groups were fed with the drugs. To observe the change of gastric ulcer of rats with cold behavior (food intake, body weight, hair color, stool, mental state); Determination of the amount of gastric juice, gastric acid, pepsin activity; to observe the pathological changes of gastric tissue HE staining; the determination of PGE2 by enzyme-linked immunosorbent assay EGF, TGF- alpha, IL-8 beta, TNF-, NT. Plasma ET, 5-HT, NE content; N0 content was determined by nitrate reduction method; Determination of AchE protein quantitative method. SPSS 19 was used for statistical treatment, P0.05 had statistical significance. Results: a cold medicine, ice acetic acid gastric ulcer was established by intragastric infusion of cold syndrome rats model of 15% acetic acid concentration had the best effect, The formation of ulcer, no fatal, and natural recovery is slow. The acetic acid filling optimum method of gastric ulcer with cold copy rat model for rats were given orally 4 C water ice Zhimu Decoction and 15% acetic acid, 1 times /d, 8:00 fed with ice Anemarrhena, 18:00 perfusion of ice acetic acid, 4 consecutive D. indoor environment setting temperature of 16 degrees Celsius, humidity 60%. two, cold efficacy of powder on gastric ulcer of rats with cold warm stomach 5 taste medicine Alpinia (1) effect on gastric ulcer of rats with cold behavioral signs: effect on body weight of gastric ulcer: comparison with cold syndrome rats the model group, 10 subjects weight rats in drug group were increased (P0.05 or P0.01). The effect on food intake of gastric ulcer of cold syndrome rats: compared with model group, 10 subjects in drug group rats increased food intake, increase rate increased significantly. The score of gastric ulcer with cold rat hair color: with the model group Comparison of 10 subjects in the drug group, low dose group galangal, rats hair color scores were lower (P0.05 or P0.01). The stool of gastric ulcer of cold syndrome rats, effects of mental status: tested drugs rats stool volume decreased, relatively dry, forming; mental state has recovery action is flexible. (two) the volume of gastric juice, gastric acid, pepsin activity effect compared with the model group, high dose of galangal Alpinia galanga, high dose, low dose of katsumada galangal, high dose, low dose of katsumada galangal fruit, high dose, low dose galanga, high dose can reduce the secretion of puzzle the gastric juice (P0.05 or P0.01); Ag high dose, high dose and low dose of Alpinia katsumada galangal,, high dose, low dose katsumada galangal fruit, high dose, Galanga low dose, high dose puzzle, the puzzle low dose group pH l hypersecretion and pepsin activity decreased (P0.0 5 or P0.01). (three) compared with the model group effect on the pathological changes of gastric tissue, reduce the infiltration of inflammatory cells in high dose group from the mucosal surface, less exudation, granulation tissue increased, mucosal surface appears new epithelial cells arranged; necrotic tissue in high dose group decreased Gaoliangjiang, muscle layer, submucosa mucosa the muscle layer structure is clear, granulation tissue increased, the mucosal surface epithelium lined up; reduce caodoukou high dose group and inflammatory cell infiltration, reduce surface exudation, granulation tissue increased, began to appear on the surface epithelial repair; reduce red nutmeg high dose group and inflammatory cell infiltration, granulation tissue distribution, mucosal surface exudate disappeared, the epithelium the organization gradually covered the mucosal surface; reduce the puzzle of high dose group and inflammatory cell infiltration, epithelial repair part of the mucosal surface; low dose group Ag mucosal exudation reduced, The granulation tissue increased, the mucosal surface of new gland cover; galangal low dose group mucosal exudation reduced, mucous layer covering new epithelial tissue; caodoukou low dose group muscle layer, submucosa structure clear, muscularis mucosa rupture began to repair, covering the surface of mucosal epithelial tissue; galanga low dose group mucosal surface exudation to reduce, reduce the infiltration of inflammatory cells, granulation tissue increased, muscularis mucosa began to repair; puzzle low dose group muscle layer structure is complete, full of submucosa granulation tissue. Three, study cold mechanism of 5 kinds of Alpinia herbs on gastric ulcer of rats with stomach cold temperature (a) compared with the model group influence on the change of PGE2 content the gastric tissue of rats with cold gastric ulcer, galangal high dose of PGE2 in gastric tissue of rats in high dose group caodoukou increased significantly, the results have significant difference (P0.01); high dose AG, katsumada Low dose, high dose galanga, PGE2 in gastric tissue in low dose group rats galanga increased significantly, the results were statistically significant (P0.05); Ag low dose, low dose and high dose of galangal, puzzle, PGE2 in gastric tissue in low dose group rat puzzle did not change significantly (P0.05). (two) effect with the model of EGF in gastric tissue of rats with gastric ulcer in the control group, the high dose of Alpinia galanga, katsumada high dose of EGF in gastric tissue of rats in high dose group puzzle was significantly increased, results there was significant difference (P0.01); high dose katsumada galangal, low dose, increase the content of EGF in gastric tissue the high dose group of galanga, the results were statistically significant (P0.05); Ag galanga low dose, low dose, no difference in intelligence gastric tissue of low dose group EGF changes (P0.05). (three) on gastric ulcer of rats with cold stomach group affected TGF-a content compared with the model group, large From high dose, low dose of galangal Alpinia galanga, high dose group, high dose of katsumada galangal fruit, high dose, puzzle of gastric tissue of rats in high dose group TGF- was significantly increased. Results there was significant difference (P0.01); low dose katsumada galangal, low dose, low dose of red cardamom, puzzle of gastric tissue low dose group rat alpha TGF- content did not change significantly (P0.05). (four) compared with the model group of gastric ulcer cold syndrome rats serum N0 content, Galanga high dose and Fuzilizhong pills, the content of serum N0 of cimetidine group rats increased significantly. Results there was significant difference (P0.01); Alpinia galanga high dose, high dose of katsumada galangal, high dose, low dose galanga, increase the content of N0 in gastric tissue of rats in high dose group puzzle, the results were statistically significant (P0.05); Ag low dose group, low dose katsumada galangal, low dose, low intelligence agent The amount of group did not change significantly (P0.05). (five) compared with the model group of gastric ulcer cold syndrome rats serum IL-8 content, katsumada galanga high dose, high dose, high dose group and the puzzle of Fuzilizhong pills, IL-8 in gastric tissue in rats of cimetidine decreased obviously, the result was significant the difference (P0.01); high dose Ag AG, low dose, high dose and low dose of Alpinia katsumada galangal, low dose, low dose, Galanga, increase the content of IL-8 in gastric tissue in low dose group rat puzzle, the results were statistically significant (P0.05). (six), cold syndrome of gastric ulcer the serum level of TNF- alpha in model group and Odaka Yoshijiangta Ag dose, low dose, high dose and low dose of Alpinia katsumada galangal,, high dose, low dose katsumada galangal fruit, high dose, low dose galanga, puzzle puzzle of high dose low dose group rats serum T There were no differences in changes of NF- content of alpha (P0.05). (seven) comparison of gastric ulcer of cold syndrome rats serum NT model group and Odaka Yoshijiangta Ag dose, low dose, high dose and low dose of Alpinia katsumada galangal,, high dose, low dose of red cardamom cardamom grass, high dose, low galanga dose puzzle high dose group rats serum NT content increased significantly (P0.01). Increased intelligence low dose group rats serum NT content, the results were statistically significant (P0.05). (eight) effect on gastric ulcer of rats with cold plasma ET content compared with the model group, high dose of katsumada galangal, high dose katsumada, low dose, high dose galanga, Galanga low dose group rat plasma ET content decreased significantly (P0.01). From high dose, low dose of galangal Alpinia galanga, low dose, high dose puzzle, reduce the content of plasma ET in the low dose group rat puzzle, statistical results Significance (P0.05). (nine) effect and model of AchE in the brain tissue of rats with gastric ulcer in the control group, the high dose of katsumada galangal, high dose, AchE content of brain tissue in rats of high dose galanga increased significantly (P0.01); Ag high dose, low dose of ginger Gao Liang increased. The content of AchE in brain tissue of rats with high dose galanga, the results were statistically significant (P0.05); Ag low dose, low dose of katsumada galangal fruit, low dose, no difference in changes of AchE content in brain tissue in rats of low dose puzzle (P0.05). (ten) effect and model of 5-HT in the brain tissue of cold rats with gastric ulcer compared to the control group, Odaka Yoshijiangta dose, high dose of katsumada galangal, high dose, high dose of galanga galanga, low dose, high dose 5-HT puzzle, in the brain tissue of low dose group of rats decreased significantly (P0.01). Educational caodoukou low dose group rat brain The contents of 5-HT decreased, the results were statistically significant (P0.05); Ag low dose group rat brain tissue from low dose of 5-HT had no significant change (P0.05). (eleven) effect and model of NE in the brain tissue of rats with gastric ulcer in the control group, katsumada high dose, high dose of red nutmeg NE, the content of brain tissue in rats of high dose significantly reduced the puzzle (P0.01). To reduce the content of NE in brain tissue of rats with low dose galanga, the results were statistically significant (P0.05); high dose Ag AG, low dose, high dose and low dose of Alpinia katsumada galangal, and low dose group the rat puzzle low dose of NE had no significant change (P0.05). Conclusion: the optimum concentration of acetic acid, acetic acid gavage type gastric ulcer in rat model of cold syndrome rats for two 15%., intragastric administration of acetic acid gastric ulcer cold syndrome and PGE2, EGF, NT, TGF- alpha, NO, decreased the content of AchE TNF-; and IL-8, ET, 5-HT, alpha, NE content increased. In three, the mechanism of 5 kinds of traditional Chinese medicine in the treatment of Alpinia acetic acid gastric ulcer with cold syndrome may reduce gastric secretion, reduce gastric acidity, pepsin activity; with the increase of PGE2, EGF, NT, NO, TGF- alpha, AchE content related; with the decrease of IL-8, plasma ET, 5-HT, NE content. Four, kinship of galangal, galangal, caodoukou, Galanga, 5 herbs in this puzzle, warm the stomach for dispelling pharmacological effects similar to the existence of the cold effect.

【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R259

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