發(fā)布時間：2018-06-09 12:04

  本文選題：辣椒素 + 胃動力　； 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：通過研究辣椒素（capsaicin，CAP）對水浸-束縛應(yīng)激（waterimmersion-restraint stress，WIRS）大鼠胃酚紅排空率的影響，檢測血漿胃動素（motilin，MTL）水平，胃竇組織辣椒素受體（transient receptor potentialvanilloid subtype1，TRPV1）、P物質(zhì)（substance P，SP）及酪氨酸激酶受體（c-kit）的表達，胃竇組織c-kit mRNA及干細胞生長因子（stem cell factor，SCF）mRNA的轉(zhuǎn)錄水平，探討CAP對大鼠胃動力的作用及機制。方法：（1）自制CAP飼料：精確稱取95％的CAP105.3mg（含CAP100mg），完全溶解于30ml的食用油中，再加入食用面粉100g，混勻，所得即為CAP含量為1mg/g的飼料。（2）WIRS大鼠模型的建立：SD大鼠20只，隨機分為正常組與模型組。模型組每日WIRS1小時，，連續(xù)4周，正常組自由進食，連續(xù)4周。（3）CAP干預(yù)實驗動物分組：SD大鼠40只，隨機分為4組：A組（正常對照組），B組（WIRS組），C組（WIRS+CAP組），D組（CAP組）。（4）具體方法：A組自由攝食進水，連續(xù)4周；B組每日WIRS1小時后自由攝食進水，連續(xù)4周；C組每日WIRS1小時后喂食CAP飼料（CAP含量為1mg／g）5g／kg／只，待CAP飼料食用完后，再給予普通飼料喂養(yǎng)，連續(xù)4周；D組每日喂食CAP飼料（CAP含量為1mg／g）5g／kg／只，待CAP飼料食用完后，再給予普通飼料喂養(yǎng)，連續(xù)4周；4周后所有大鼠禁食24小時，于次日早晨給予濃度為50mg／dL的酚紅溶液灌胃，2ml/鼠，灌胃30min后麻醉處死大鼠。（5）檢測指標：①收集胃內(nèi)容物，測定胃酚紅排空率。②腹主動脈取血2ml，4℃離心后提取上清液，用Elisa法檢測血漿MTL水平。③肉眼觀察大鼠胃粘膜損傷指數(shù)并進行病理組織學(xué)檢查。④取胃竇組織包埋、切片后用免疫組化方法檢測TRPV1、SP及c-kit的表達。⑤取胃竇組織用RT-PCR方法檢測c-kit mRNA、SCF mRNA的轉(zhuǎn)錄水平。結(jié)果：1.模型驗證：（1）動物一般情況：實驗過程中，正常組大鼠一般狀態(tài)好，活動靈活，進食正常，大小便正常，無死亡；模型組大鼠精神狀態(tài)差，行動遲緩，進食較少，大便干結(jié)，但無死亡。（2）大鼠胃酚紅排空率：正常組、WIRS模型組大鼠胃酚紅排空率分別為：61.76%±1.22%，53.07%±2.19%，WIRS模型組胃酚紅排空率顯著低于正常組（P＜0.05）。2. CAP干預(yù)的結(jié)果：（1）動物一般情況：實驗過程中各組大鼠無死亡，其中A組大鼠一般狀態(tài)好，活動靈活，進食正常，大小便正常；B組大鼠精神狀態(tài)差，行動遲緩，進食較少，大便干結(jié)；C組大鼠一般狀態(tài)良好，活動稍遲緩，進食正常，大小便正常；D組大鼠一般狀態(tài)好，活動靈活，進食較多，大小便正常。（2）大鼠胃酚紅排空率：A組62.91%±1.10%，B組55.33%±1.79%，C組61.88%±2.07%，D組70.78%±2.42%，其中B組胃酚紅排空率顯著低于A組、C組、D組（P＜0.05），D組胃酚紅排空率顯著高于A組（P＜0.05），A組胃酚紅排空率與C組比較差異無統(tǒng)計學(xué)意義（P＞0.05）。（3）大鼠血漿MTL水平：A組190.30±1.68pg/ml，B組179.75±1.72pg/ml，C組200.06±2.06pg/ml，D組201.37±1.45pg/ml，其中B組血漿MTL水平顯著低于A組、C組及D組（P＜0.05），A組血漿MTL水平顯著低于C組、D組（P＜0.05），C組血漿MTL水平與D組比較差異無統(tǒng)計學(xué)意義（P＞0.05）。（4）免疫組化方法檢測結(jié)果：①大鼠胃竇組織TRPV1表達積分結(jié)果：A組2.30±0.48分，B組2.20±0.42分，C組3.30±0.48分，D組3.60±0.52分，其中A組TRPV1表達水平顯著低于C組、D組（P＜0.05），B組TRPV1表達水平顯著低于C組及D組（P＜0.05），A組TRPV1表達水平與B組差異無統(tǒng)計學(xué)意義（P＞0.05），C組TRPV1表達水平與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。②大鼠胃竇組織SP表達積分結(jié)果：A組為2.00±0.47分，B組為1.20±0.42分，C組為3.20±0.63分，D組為3.50±0.53分，其中B組SP表達水平顯著低于A組、C組及D組（P＜0.05），A組SP表達水平顯著低于C組及D組（P＜0.05），C組SP表達水平與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。③大鼠胃竇組織c-kit表達積分結(jié)果：A組1.30±0.48分，B組0.80±0.42分，C組1.50±0.53分，D組1.70±0.48分，其中B組c-kit表達水平顯著低于A組、D組（P＜0.05），A組c-kit表達水平與C組、D組差異無統(tǒng)計學(xué)意義（P＞0.05），B組c-kit表達水平與C組差異無統(tǒng)計學(xué)意義（P＞0.05），C組c-kit表達水平與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。（5）RT-PCR方法檢測結(jié)果：①大鼠胃竇組織c-kit mRNA表達量結(jié)果：A組為0.624±0.016分，B組為0.606±0.011分，C組為0.622±0.011分，D組為0.633±0.013分，其中B組c-kitmRNA轉(zhuǎn)錄水平顯著低于A組、D組（P＜0.05），A組c-kit mRNA轉(zhuǎn)錄水平與C組、D組差異無統(tǒng)計學(xué)意義（P＞0.05），B組c-kit mRNA轉(zhuǎn)錄水平與C組差異無統(tǒng)計學(xué)意義（P＞0.05），C組c-kit mRNA轉(zhuǎn)錄水平與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。②大鼠胃竇組織SCF mRNA表達量結(jié)果：A組為0.894±0.011分，B組為0.853±0.009分，C組為0.932±0.009分，D組為0.949±0.012分，其中B組SCF mRNA轉(zhuǎn)錄水平顯著低于A組、C組及D組（P＜0.05），A組SCF mRNA轉(zhuǎn)錄水平顯著低于C組、D組（P＜0.05），C組SCF mRNA轉(zhuǎn)錄水平與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。（6）胃粘膜損傷指數(shù)及病理組織學(xué)檢查結(jié)果：①胃粘膜損傷評分（Guth標準改良評分）：A組0.40±0.52分，B組2.90±0.74分，C組1.40±0.52分，D組0.50±0.53分，其中B組胃粘膜損傷評分顯著高于A組、C組及D組（P＜0.05），C組胃粘膜損傷評分顯著高于A組、D組（P＜0.05），A組胃粘膜損傷評分與D組差異無統(tǒng)計學(xué)意義（P＞0.05）②胃粘膜病理組織損傷積分（Masuda標準方法）：A組0.30±0.48分，B組3.70±1.89分，C組1.60±0.70分，D組0.40±0.52分，其中B組病理組織損傷積分顯著高于A組、C組及D組（P＜0.05），C組病理組織損傷積分顯著高于A組、D組（P＜0.05），A組病理組織損傷積分與D組差異無統(tǒng)計學(xué)意義（P＞0.05）。結(jié)論：（1）WIRS能成功建立胃動力障礙動物模型。（2）正常大鼠攝食小劑量CAP飼料4周可促進胃動力。（3）WIRS大鼠攝食小劑量CAP飼料4周可改善胃動力。（4）CAP影響正常及WIRS大鼠的胃動力可能與CAP調(diào)節(jié)TRPV1、SP的表達及MTL的釋放有關(guān)。（5）CAP可能對SCF的表達有一定影響，但與c-kit、Cajal間質(zhì)細胞的相關(guān)性尚不明確。
[Abstract]:Objective: To investigate the effect of capsaicin (CAP) on the gastric emptying rate of waterimmersion-restraint stress (WIRS) rats, and to detect the level of motilin (MTL), capsaicin receptor (transient receptor potentialvanilloid subtype1, TRPV1), and tyrosine in the gastric antrum tissue. The expression of kinase receptor (c-kit), the transcriptional level of c-kit mRNA and stem cell growth factor (stem cell factor, SCF) mRNA in the gastric antrum to explore the effect and mechanism of CAP on gastric motility in rats. Methods: (1) the self-made CAP feed: exactly 95% CAP105.3mg (containing CAP100mg), completely dissolved in edible oil, and then added to edible flour, (2) the model of WIRS rats was established: (2) the model of WIRS rats was established: 20 SD rats were randomly divided into normal group and model group. The model group was WIRS1 hours a day for 4 weeks. The normal group was free to eat for 4 weeks. (3) CAP intervention experimental animals were divided into 4 groups of SD rats randomly, A group (normal control group), B group (WIRS group), C Group (group WIRS+CAP), group D (group CAP). (4) specific methods: group A was free to feed water for a continuous period of 4 weeks; group B was free to feed water after WIRS1 hours a day for 4 weeks; C group was fed CAP feed after WIRS1 hours a day (CAP content was 1mg / g). The feed (CAP content was 1mg / g) 5g / kg / only, after the CAP feed was finished, the feed was given to the ordinary feed for 4 weeks. After 4 weeks, all rats were fasted for 24 hours, and the rats were given a concentration of 50mg / dL in the next morning. The rats were sacrificed after the stomach 30min. (5) the contents of gastric contents were collected and the gastric phenol red was measured. The rate of evacuation of the abdominal aorta was 2ml, the supernatant was extracted after 4 degrees centigrade centrifugation and the plasma MTL level was detected by Elisa. (3) the gastric mucosa injury index of the rats was observed by the naked eye and the histopathological examination was performed. 4. The gastric antrum tissue was embedded and the expression of TRPV1, SP and c-kit was detected by immunohistochemical method. RT-PCR method was used to detect c-k in the gastric antrum tissue. It mRNA, SCF mRNA transcriptional level. Results: 1. model validation: (1) animal general condition: in the course of the experiment, normal group rats have good general state, flexible activity, normal eating, normal size and stool, no death; the model group rats have poor mental state, slow action, eating less, stool dry, but no death. (2) gastric emptying rate of stomach in rats: (2) positive gastric emptying rate: positive stomach rats: positive gastric phenol red evacuation rate: positive stomach rat gastric phenol red evacuation rate: positive rat stomach phenol red evacuation rate: positive rate of gastric phenol red: positive rat stomach phenol red evacuation rate: positive rate of gastric phenol red: positive rat stomach phenol red evacuation rate: positive rate of gastric phenol red: positive rat stomach phenol red evacuation rate: Zheng Zheng The rate of gastric phenol red emptying in the WIRS model group was 61.76% + 1.22% and 53.07% + 2.19% respectively. The gastric phenol red emptying rate in the WIRS model group was significantly lower than that of the normal group (P < 0.05).2. CAP intervention: (1) the animal general situation: the rats in each group were not dead, of which group A rats were in good condition, flexible activities, eating normal, stool and stool Normal, group B rats were poor in mental state, slow action, less eating and stool; group C rats were generally in good condition, slow activity, normal eating, normal size and stool; group D rats had good general state, flexible activity, more eating, and normal size and stool. (2) the rate of gastric emptying in the stomach of rats was 62.91% + 1.10%, B group 55.33% + 1.79%, and C group 61.88% + 2 .07% and group D were 70.78% + 2.42%. The rate of gastric phenol red emptying in group B was significantly lower than that in group A, C group, D group (P < 0.05), and the rate of gastric phenol red emptying in D group was significantly higher than that in group A (P < 0.05), and there was no significant difference between A group and C group. (3) the plasma levels of rats were 190.30 and 179.75. Group D was 201.37 + 1.45pg/ml, and the plasma MTL level in group B was significantly lower than that in group A, C group and D group (P < 0.05). The plasma MTL level of A group was significantly lower than that in C group (< 0.05). (4) there was no significant difference between the group and group (4) immunohistochemical method test results: (1) the result of the expression score of the gastric antrum tissue of the rat: 2.30 + 0 in the group of rats .48 score, group B was 2.20 + 0.42, group C was 3.30 + 0.48, and D group was 3.60 + 0.52. The TRPV1 expression level in A group was significantly lower than that in C group, D group (P < 0.05), TRPV1 expression level in B group was significantly lower than that in C group and group (0.05 < 0.05). The results of SP expression in the gastric antrum of rats were 2 + 0.47 points, B group 1.20 + 0.42, C group 3.20 + 0.63, D group 3.50 + 0.53, B group SP expression level was significantly lower than that of A group, C group and D group (P < 0.05), A group expression level was significantly lower than that of 0.05 group (0.05). 0.05). The results of c-kit expression in the gastric antrum of rats were 1.30 + 0.48 points, group B 0.80 + 0.42, C group 1.50 + 0.53, D group 1.70 + 0.48, c-kit expression level in group B was significantly lower than that in A group, D group (P < 0.05), A group c-kit expression level was not statistically significant (0.05). Significance (P > 0.05), the expression level of c-kit in group C was not statistically significant (P > 0.05). (5) RT-PCR method detection results: (1) the expression of c-kit mRNA in the gastric antrum of rats: A group was 0.624 + 0.016, B group was 0.606 + 0.011, C group was 0.622 + 0.011, D group was 0.633 + 0.013, and the transcriptional level of the group was significantly lower than that of the group. D group (P < 0.05), c-kit mRNA transcriptional level in group A and C group, D group had no statistical significance (P > 0.05), c-kit mRNA transcriptional level in group B was not statistically significant (> 0.05). 0.853 + 0.009 points, C group was 0.932 + 0.009, and D group was 0.949 + 0.012. The SCF mRNA transcriptional level in group B was significantly lower than that in A group, C group and D group (P < 0.05), SCF mRNA in group A group was significantly lower than that of group (6). (6) gastric mucosal injury index and histopathology The results of examination: (1) gastric mucosal injury score (Guth standard improvement score) 0.40 + 0.52 points in group:A, 2.90 + 0.74 in group B, 1.40 + 0.52 in group C, 0.50 in group D in group B, significantly higher than that in group A, C and D group (P < 0.05), and C group gastric mucosal injury score was significantly higher than that in A group (0.05). The difference was not statistically significant (P > 0.05). The score of Gastric Mucosal pathological tissue injury (Masuda standard method) was 0.30 + 0.48 points in group:A, 3.70 + 1.89 in group B, 1.60 + 0.70 in group C, and 0.40 + 0.52 in group D, among which, pathological tissue injury scores in group B were significantly higher than that in A group, C group and D group (P < 0.05). There was no significant difference between the injury score of pathological tissue and D group (P > 0.05). Conclusion: (1) WIRS could successfully establish the animal model of gastric motility disorder. (2) normal rats fed with small dose CAP feed for 4 weeks could promote gastric motility. (3) WIRS rats fed a small dose of CAP feed for 4 weeks to improve gastric motility. (4) CAP affects the gastric motility of normal and WIRS rats. It is related to CAP regulating the expression of TRPV1, SP and the release of MTL. (5) CAP may have an effect on the expression of SCF, but the correlation with c-kit and Cajal interstitial cells is not clear.
【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】：2014
【分類號】：R285.5

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前1條

1 李毅;齊清會;;胃腸道神經(jīng)-Cajal間質(zhì)細胞-平滑肌網(wǎng)絡(luò)研究進展[J];國際消化病雜志;2007年03期

本文編號：1999661