發(fā)布時間：2018-06-11 19:54

  本文選題：辣椒素 + 內(nèi)臟高敏感��； 參考：《瀘州醫(yī)學(xué)院》2010年碩士論文

【摘要】： 目的：通過觀察不同劑量的辣椒素(capsaicin, CAP)對內(nèi)臟高敏感大鼠胃酸分泌、胃底腺區(qū)組織內(nèi)辣椒素受體(vanilloid receptor subtype 1, VR1)及降鈣素基因相關(guān)肽(calcitonin gene-related peptide, CGRP)和P物質(zhì)(substance P, SP)、血漿胃泌素(GAS)和胃粘膜屏障的影響,探討不同劑量的CAP對內(nèi)臟高敏感大鼠胃酸的影響及其機(jī)制。方法：(1)實(shí)驗(yàn)動物分組：60只SD大鼠,雌雄不拘,體重180g～200g之間,隨機(jī)分為A組(對照組)、B組(模型組)、C1組(CAP1組)、C2組(CAP2組)及C3組(CAP3組),每組大鼠12只。(2)內(nèi)臟高敏感大鼠模型的建立：實(shí)驗(yàn)大鼠適應(yīng)性飼養(yǎng)2w,自由進(jìn)食進(jìn)水。造模方法：A組腹腔注射生理鹽水,1ml/鼠；B組、C1組、C2組和C3組大鼠腹腔注射雞卵清白蛋白(chicken egg albumin), 1ml/鼠,制備內(nèi)臟高敏感大鼠模型。造模過程中觀察大鼠一般情況。2w后取A組全部大鼠12只組成正常對照組,隨機(jī)從B組、C1組、C2組和C3組中抽取共15只大鼠組成模型對照組采用肛門直腸擴(kuò)張(colorectal distention, CRD)實(shí)驗(yàn)以觀察大鼠腹部撤離反射(abdominal withdrawal reflex, AWR),并用AWR評分評估大鼠對直腸擴(kuò)張刺激的內(nèi)臟感覺改變。(3)CAP的使用方法及劑量：大鼠造模成功后進(jìn)行CAP治療實(shí)驗(yàn),具體方法如下：A組：自由進(jìn)食進(jìn)水,生理鹽水10mL/kg/d分2次灌胃,連續(xù)2w；B組：自由進(jìn)食進(jìn)水,生理鹽水10mL/kg/d分2次灌胃,連續(xù)2w；C1組：自由進(jìn)食進(jìn)水,CAP 1mg/kg/d分2次灌胃灌胃,容積為10mL/kg/d,連續(xù)2w；C2組：自由進(jìn)食進(jìn)水,CAP 5mg/kg/d分2次灌胃灌胃,灌胃容積為10mL/kg/d,連續(xù)2w；C3組：自由進(jìn)食進(jìn)水,CAP20mg/kg/d分2次灌胃,灌胃容積為10mL/kg/d,連續(xù)2w。(4)檢測指標(biāo)及方法：①造模及給藥過程中觀察大鼠的一般情況。②胃酸的測定：所有大鼠于實(shí)驗(yàn)前一夜開始禁食(不禁水),末次灌胃后1h,用質(zhì)量分?jǐn)?shù)為2%的戊巴比妥鈉(40mg/kg)腹腔注射麻醉,打開腹腔,結(jié)扎食管胃連接部和幽門部,將整個胃取出,剪開胃并用生理鹽水清洗胃內(nèi)容物,收集清洗液,用酸堿滴定法測定胃酸。③胃底腺區(qū)組織內(nèi)VR1和CGRP、SP的表達(dá)情況：取各組大鼠胃底腺區(qū)組織(約0.5cm×0.5cm)全層一塊,立刻置于4%的多聚甲醛中充分固定2h,石蠟包埋,連續(xù)5μm切片,用免疫組化方法分別進(jìn)行CGRP、SP、VR1測定。④血漿GAS：心臟采血2ml,4℃離心后提取上清液,用放射免疫法檢測血漿GAS水平的變化。⑤胃粘膜屏障改變：把清洗過胃內(nèi)容物之后的胃置于4%的多聚甲醛中充分固定后,肉眼觀察胃粘膜形態(tài),采用Guth標(biāo)準(zhǔn)改良評分評定胃粘膜損傷情況,并取胃粘膜組織送病理組織學(xué)檢查,光鏡下對HE染色后的胃粘膜進(jìn)行觀察。結(jié)果：(1)動物一般情況：造模過程中,A組大鼠精神狀態(tài)好,行動靈活,食欲佳,皮毛光澤,大小便正常,體重增加；B組、C1組、C2組、C3組大鼠出現(xiàn)好斗,易激惹,互相撕咬等情況,但無死亡。隨著CAP治療時間延長,C1組、C2組、C3組大鼠好斗、激惹、互相撕咬情況減少,而B組大鼠無明顯減少。在灌胃過程中C1組有5只大鼠、C2組有5只大鼠、C3組有6只大鼠曾出現(xiàn)嗆咳,但均無窒息和死亡。(2)AWR評分結(jié)果：正常對照組注水1ml、3ml和5ml的AWR評分分別為：0.67±0.50分、1.44±0.53分、2.56±0.53分,模型對照組注水1ml、3ml和5ml的AWR評分分別為：0.53±0.52分、2.53±0.52分、3.40±0.63分。隨著氣囊體積的增加,正常對照組和模型對照組AWR評分值逐漸遞增。模型對照組注水3ml和5ml氣囊擴(kuò)張時AWR評分均顯著高于正常對照組(P0.05)。(3)胃液總酸度的變化：各組大鼠的胃液總酸度分別為：A組48.17±5.75mmol/L,B組45.67±3.05 mmol/L,C1組45.25±2.93 mmol/L,C2組34.42±2.28 mmol/L,C3組30.50±4.25 mmol/L.其中B組大鼠胃液總酸度低于A組大鼠,但沒有統(tǒng)計(jì)學(xué)意義(P0.05)；C1組大鼠胃液總酸度和B組大鼠胃液總酸度比較沒有統(tǒng)計(jì)學(xué)意義(P0.05)；隨著CAP劑量增加。,C2組大鼠胃液總酸度顯著低于A組(P0.05),C2組大鼠胃液總酸度顯著低于B組(P0.05),C2組大鼠胃液總酸度顯著低于C1組(P0.05)；并且隨著CAP劑量繼續(xù)增加,C3組大鼠胃液總酸度顯著低于A組(P0.05),C3組大鼠胃液總酸度顯著低于B組(P0.05),C3組大鼠胃液總酸度顯著低于C1組(P0.05),C3組大鼠胃液總酸度顯著低于C2組(P0.05)。(4)胃底腺區(qū)組織內(nèi)VR1的表達(dá)情況：大鼠胃底腺區(qū)組織內(nèi)VRl表達(dá)水平分別為：A組為0.58±0.51分,B組為1.25±0.62分,C1組為2.00±0.60分,C2組為2.58±0.52分,C3組為3.25±0.75分,其中A組VR1表達(dá)水平顯著低于B組(P0.05),A組VR1表達(dá)水平顯著低于C1組(P0.05),A組VR1表達(dá)水平顯著低于C2組(P0.05),A組VR1表達(dá)水平顯著低于C3組(P0.05)；B組VR1表達(dá)水平也顯著低于C1組(P0.05),B組VR1表達(dá)水平也顯著低于C2組(P0.05),B組VR1表達(dá)水平也顯著低于C3組(P0.05)；隨著CAP劑量的增加,C2組VR1表達(dá)水平顯著高于C1組(P0.05)；C3組VR1水平也顯著高于C2組(P0.05)。(5)胃底腺區(qū)組織內(nèi)CGRP.SP的表達(dá)情況：大鼠胃底腺區(qū)組織內(nèi)CGRP表達(dá)水平分別為：A組為1.00±0.43分,B組為1.08±0.52分,C1組為1.58±0.52分,C2組為2.17±0.58分,C3組為2.67±0.49分,其中A組CGRP表達(dá)水平與B組相比無顯著性差異(P0.05),A組CGRP表達(dá)水平顯著低于C1組(P0.05),A組CGRP表達(dá)水平顯著低于C2組(P0.05),A組CGRP表達(dá)水平顯著低于C3組(P0.05)；B組CGRP表達(dá)水平也顯著低于C1組(P0.05),B組CGRP表達(dá)水平也顯著低于C2組(P0.05),B組CGRP表達(dá)水平也顯著低于C3組(P0.05)；隨著CAP劑量的增加,C2組CGRP表達(dá)水平顯著高于C1組(P0.05),C3組CGRP水平也顯著高于C2組(P0.05)。大鼠胃底腺區(qū)組織內(nèi)SP表達(dá)水平分別為：A組為0.50±0.52分, B組為0.83±0.58分,C1組為0.75±0.45分,C2組為0.75±0.45分,C3組為0.58±0.52分,其中B組SP表達(dá)水平較A組略有升高,但沒有統(tǒng)計(jì)學(xué)意義(P0.05)；其余各組間SP表達(dá)水平均沒有統(tǒng)計(jì)學(xué)意義。(6)血漿GAS的變化：大鼠血漿GAS水平分別為：A組49.50±7.51 pg/ml,B組63.25±5.99 pg/ml,C1組44.33±2.90 pg/ml,C2組37.42±3.55 pg/ml,C3組33.00±2.37pg/ml。其中A組GAS水平顯著低于B組(P0.05),A組GAS高于C1組(P0.05),A組GAS高于C2組(P0.05),A組GAS低于C3組(P0.05)；B組GAS水平也顯高于C1組(P0.05),B組GAS水平也顯高于C2組(P0.05),B組GAS水平也顯高于C3組(P0.05)；C2組GAS水平相比顯著低于C1組(P0.05),而C3組的血漿GAS水平顯著低于C2組(P0.05)。(7)胃粘膜屏障：A組、B組、C1組、C2組及C3組的胃粘膜損傷評分(Guth標(biāo)準(zhǔn)改良評分)比較各組間無顯著性差異(P0.05)；光鏡下觀察胃粘膜組織病理學(xué)改變：A組、B組、C1組、C2組大鼠胃粘膜基本光滑,上皮細(xì)胞形態(tài)正常,排列整齊,未見粘膜脫落、充血、水腫、炎性細(xì)胞浸潤及腺體結(jié)構(gòu)紊亂和壞死。C3組大鼠胃粘膜上皮細(xì)胞輕度脫落、充血、水腫,未見腺體結(jié)構(gòu)紊亂和壞死。對胃粘膜病理損傷程度進(jìn)行半定量統(tǒng)計(jì),病理損傷積分A組為0.25±0.45分,B組為0.17±0.39分,C1組為0.32±0.45分,C2組為0.33±0.47分,C3組為0.83±0.72分。其中C3組病理損傷積分顯著高于A組(P0.05),C3組病理損傷積分顯著高于B組(P0.05),C3組病理損傷積分顯著高于C1組(P0.05),C3組病理損傷積分顯著高于C2組(P0.05)。結(jié)論：1.腹腔注射雞卵清白蛋白可成功建立大鼠內(nèi)臟高敏感模型。2.CAP1mg/kgd、5mg /kg/d及20mg/kg/d連續(xù)灌胃2w可抑制內(nèi)臟高敏感大鼠胃酸分泌,并且隨著灌胃劑量的增加,抑酸效應(yīng)增強(qiáng)。3.CAP影響內(nèi)臟高敏感大鼠的胃酸分泌可能通過激活胃壁內(nèi)VRl而發(fā)揮作用。4.CAP影響內(nèi)臟高敏感大鼠的胃酸分泌可能與CGRP有關(guān),與SP無明顯相關(guān)。5.CAP影響內(nèi)臟高敏感大鼠的胃酸分泌可能與GAS有關(guān)6. CAP 1mg/kg/d和5mg/kg/d連續(xù)灌胃對正常和內(nèi)臟高敏感大鼠的胃粘膜屏障無損傷作用,而CAP 20mg/kg/d連續(xù)灌胃2w可造成胃粘膜輕微的損傷。
[Abstract]:Objective : To investigate the effects of different doses of capsaicin ( CAP ) on gastric acid secretion , gastrin ( GAS ) and gastric mucosal barrier in rats with visceral hypersensitivity . Methods : ( 1 ) Group B ( model group ) , group C1 ( CAP1 group ) , C2 group ( CAP2 group ) and C3 group ( CAP3 group ) were randomly divided into group A ( control group ) , group B ( model group ) , C1 group ( CAP1 group ) , C2 group ( CAP2 group ) and C3 group ( CAP3 group ) , and the rats were randomly divided into group A ( control group ) , group B ( model group ) , group C1 ( CAP1 group ) , C2 group ( CAP2 group ) and C3 group ( CAP3 group ) .
The rats were randomly divided into group B , C1 , C2 and C3 . The rats were randomly divided into group B , C1 , C2 and C3 . The rats were randomly divided into group B , C1 , C2 and C3 . The rats were randomly divided into group B , C1 , C2 and C3 .
Group B : Free food intake , normal saline 10 mL / kg / day divided into 2 times , continuous 2w ;
C1 group : free food intake , CAP 1mg / kg / day divided into two times , the volume was 10 mL / kg / d , continuous 2w ;
Group 2 : Free food intake , CAP 5mg / kg / day divided into two times , the volume of gastric lavage was 10 mL / kg / d , continuous 2w ;
The gastric mucosal barrier was determined by using the Guth standard modified score to assess the gastric mucosa injury .
The AWR scores of group B , C1 , C2 , C3 were significantly higher than those in normal control group ( P 0.05 ) .
There was no significant difference between the total acidity of gastric juice and the total acidity of gastric juice in group C ( P0.05 ) .
The total acidity of gastric juice in C2 group was significantly lower than that in group A ( P0.05 ) , and the total acidity of gastric juice in C2 group was significantly lower than that in group C ( P0.05 ) .
The total acidity of gastric juice in group C was significantly lower than that in group C ( P0.05 ) . The total acidity of gastric juice in group C was significantly lower than that in group C ( P0.05 ) .
The level of expression in group B was also significantly lower than that in group C ( P0.05 ) , and the level of expression in group B was significantly lower than that in group C ( P0.05 ) , and the level of expression in group B was significantly lower than that in group C ( P0.05 ) .
With the increase of CAP dose , the level of expression level in C2 group was significantly higher than that in C1 group ( P0.05 ) .
The level of CGRP in group C was significantly higher than that in group C ( P0.05 ) . The level of CGRP in group A was 1.00 鹵 0.43 , group B was 1.08 鹵 0.52 , group C was 1 . 58 鹵 0.52 , group C was 2 . 17 鹵 0.58 , C3 group was 2.67 鹵 0.49 , group A was significantly lower than that in group C ( P0.05 ) , CGRP expression level in group A was significantly lower than that in group C ( P0.05 ) .
The level of CGRP in group B was significantly lower than that in group C ( P0.05 ) , and the level of CGRP in group B was significantly lower than that in group C ( P0.05 ) .
The level of CGRP in group C was significantly higher than that in group C ( P0.05 ) . The levels of CGRP in group C were 0.50 鹵 0.52 , 0.83 鹵 0.58 in group A , 0.75 鹵 0.45 in group C , 0.75 鹵 0.45 in group C , 0.58 鹵 0.52 in group C , and 0.58 鹵 0.52 in group C , but not statistically significant ( P0.05 ) .
The levels of GAS in plasma GAS were 49.50 鹵 7.51 pg / ml , 63.25 鹵 5.99 pg / ml in group B , 44.33 鹵 2.90 pg / ml in group B , 37.42 鹵 3.55 pg / ml in group C , 30.00 鹵 2.37pg / ml in C3 group .
The GAS levels in group B were also significantly higher than those in group C ( P0.05 ) , and GAS levels in group B were significantly higher than those in group C ( P0.05 ) .
The level of GAS in group C was significantly lower than that in group C ( P0.05 ) , while the level of plasma GAS in C3 group was significantly lower than that in group C ( P0.05 ) . ( 7 ) There was no significant difference between group A , group B , C1 , C2 and C3 group ( P0.05 ) .
The pathological changes of gastric mucosa in C3 group were significantly higher than those in group A ( P0.05 ) . CAP 1mg / kg / day and 5mg / kg / d continuous gastric lavage had no damage to the gastric mucosa barrier of rats with normal and visceral hypersensitivity , while CAP 20mg / kg / day could cause slight injury to gastric mucosa .
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 蔣海清,侯奕,黃曉焰,謬作華,孫慶偉;辣椒治療胃粘膜損傷及潰瘍的實(shí)驗(yàn)與臨床研究[J];贛南醫(yī)學(xué)院學(xué)報;2003年04期

2 徐晶華;;辣椒辣素在胃損傷和治愈過程中的作用[J];黑龍江醫(yī)藥;2003年05期

3 王健平,朱文玉,洪昭雄,湯健,吳t，

本文編號：2006500