辣椒素对水浸—束缚应激大鼠胃动力的作用及机制研究

发布时间：2018-06-09 12:04

  本文选题：辣椒素 + 胃动力　； 参考：《泸州医学院》2014年硕士论文

【摘要】：目的：通过研究辣椒素（capsaicin，CAP）对水浸-束缚应激（waterimmersion-restraint stress，WIRS）大鼠胃酚红排空率的影响，检测血浆胃动素（motilin，MTL）水平，胃窦组织辣椒素受体（transient receptor potentialvanilloid subtype1，TRPV1）、P物质（substance P，SP）及酪氨酸激酶受体（c-kit）的表达，胃窦组织c-kit mRNA及干细胞生长因子（stem cell factor，SCF）mRNA的转录水平，探讨CAP对大鼠胃动力的作用及机制。方法：（1）自制CAP饲料：精确称取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，混匀，所得即为CAP含量为1mg/g的饲料。（2）WIRS大鼠模型的建立：SD大鼠20只，随机分为正常组与模型组。模型组每日WIRS1小时，，连续4周，正常组自由进食，连续4周。（3）CAP干预实验动物分组：SD大鼠40只，随机分为4组：A组（正常对照组），B组（WIRS组），C组（WIRS+CAP组），D组（CAP组）。（4）具体方法：A组自由摄食进水，连续4周；B组每日WIRS1小时后自由摄食进水，连续4周；C组每日WIRS1小时后喂食CAP饲料（CAP含量为1mg／g）5g／kg／只，待CAP饲料食用完后，再给予普通饲料喂养，连续4周；D组每日喂食CAP饲料（CAP含量为1mg／g）5g／kg／只，待CAP饲料食用完后，再给予普通饲料喂养，连续4周；4周后所有大鼠禁食24小时，于次日早晨给予浓度为50mg／dL的酚红溶液灌胃，2ml/鼠，灌胃30min后麻醉处死大鼠。（5）检测指标：①收集胃内容物，测定胃酚红排空率。②腹主动脉取血2ml，4℃离心后提取上清液，用Elisa法检测血浆MTL水平。③肉眼观察大鼠胃粘膜损伤指数并进行病理组织学检查。④取胃窦组织包埋、切片后用免疫组化方法检测TRPV1、SP及c-kit的表达。⑤取胃窦组织用RT-PCR方法检测c-kit mRNA、SCF mRNA的转录水平。结果：1.模型验证：（1）动物一般情况：实验过程中，正常组大鼠一般状态好，活动灵活，进食正常，大小便正常，无死亡；模型组大鼠精神状态差，行动迟缓，进食较少，大便干结，但无死亡。（2）大鼠胃酚红排空率：正常组、WIRS模型组大鼠胃酚红排空率分别为：61.76%±1.22%，53.07%±2.19%，WIRS模型组胃酚红排空率显著低于正常组（P＜0.05）。2. CAP干预的结果：（1）动物一般情况：实验过程中各组大鼠无死亡，其中A组大鼠一般状态好，活动灵活，进食正常，大小便正常；B组大鼠精神状态差，行动迟缓，进食较少，大便干结；C组大鼠一般状态良好，活动稍迟缓，进食正常，大小便正常；D组大鼠一般状态好，活动灵活，进食较多，大小便正常。（2）大鼠胃酚红排空率：A组62.91%±1.10%，B组55.33%±1.79%，C组61.88%±2.07%，D组70.78%±2.42%，其中B组胃酚红排空率显著低于A组、C组、D组（P＜0.05），D组胃酚红排空率显著高于A组（P＜0.05），A组胃酚红排空率与C组比较差异无统计学意义（P＞0.05）。（3）大鼠血浆MTL水平：A组190.30±1.68pg/ml，B组179.75±1.72pg/ml，C组200.06±2.06pg/ml，D组201.37±1.45pg/ml，其中B组血浆MTL水平显著低于A组、C组及D组（P＜0.05），A组血浆MTL水平显著低于C组、D组（P＜0.05），C组血浆MTL水平与D组比较差异无统计学意义（P＞0.05）。（4）免疫组化方法检测结果：①大鼠胃窦组织TRPV1表达积分结果：A组2.30±0.48分，B组2.20±0.42分，C组3.30±0.48分，D组3.60±0.52分，其中A组TRPV1表达水平显著低于C组、D组（P＜0.05），B组TRPV1表达水平显著低于C组及D组（P＜0.05），A组TRPV1表达水平与B组差异无统计学意义（P＞0.05），C组TRPV1表达水平与D组差异无统计学意义（P＞0.05）。②大鼠胃窦组织SP表达积分结果：A组为2.00±0.47分，B组为1.20±0.42分，C组为3.20±0.63分，D组为3.50±0.53分，其中B组SP表达水平显著低于A组、C组及D组（P＜0.05），A组SP表达水平显著低于C组及D组（P＜0.05），C组SP表达水平与D组差异无统计学意义（P＞0.05）。③大鼠胃窦组织c-kit表达积分结果：A组1.30±0.48分，B组0.80±0.42分，C组1.50±0.53分，D组1.70±0.48分，其中B组c-kit表达水平显著低于A组、D组（P＜0.05），A组c-kit表达水平与C组、D组差异无统计学意义（P＞0.05），B组c-kit表达水平与C组差异无统计学意义（P＞0.05），C组c-kit表达水平与D组差异无统计学意义（P＞0.05）。（5）RT-PCR方法检测结果：①大鼠胃窦组织c-kit mRNA表达量结果：A组为0.624±0.016分，B组为0.606±0.011分，C组为0.622±0.011分，D组为0.633±0.013分，其中B组c-kitmRNA转录水平显著低于A组、D组（P＜0.05），A组c-kit mRNA转录水平与C组、D组差异无统计学意义（P＞0.05），B组c-kit mRNA转录水平与C组差异无统计学意义（P＞0.05），C组c-kit mRNA转录水平与D组差异无统计学意义（P＞0.05）。②大鼠胃窦组织SCF mRNA表达量结果：A组为0.894±0.011分，B组为0.853±0.009分，C组为0.932±0.009分，D组为0.949±0.012分，其中B组SCF mRNA转录水平显著低于A组、C组及D组（P＜0.05），A组SCF mRNA转录水平显著低于C组、D组（P＜0.05），C组SCF mRNA转录水平与D组差异无统计学意义（P＞0.05）。（6）胃粘膜损伤指数及病理组织学检查结果：①胃粘膜损伤评分（Guth标准改良评分）：A组0.40±0.52分，B组2.90±0.74分，C组1.40±0.52分，D组0.50±0.53分，其中B组胃粘膜损伤评分显著高于A组、C组及D组（P＜0.05），C组胃粘膜损伤评分显著高于A组、D组（P＜0.05），A组胃粘膜损伤评分与D组差异无统计学意义（P＞0.05）②胃粘膜病理组织损伤积分（Masuda标准方法）：A组0.30±0.48分，B组3.70±1.89分，C组1.60±0.70分，D组0.40±0.52分，其中B组病理组织损伤积分显著高于A组、C组及D组（P＜0.05），C组病理组织损伤积分显著高于A组、D组（P＜0.05），A组病理组织损伤积分与D组差异无统计学意义（P＞0.05）。结论：（1）WIRS能成功建立胃动力障碍动物模型。（2）正常大鼠摄食小剂量CAP饲料4周可促进胃动力。（3）WIRS大鼠摄食小剂量CAP饲料4周可改善胃动力。（4）CAP影响正常及WIRS大鼠的胃动力可能与CAP调节TRPV1、SP的表达及MTL的释放有关。（5）CAP可能对SCF的表达有一定影响，但与c-kit、Cajal间质细胞的相关性尚不明确。
[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.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5

【参考文献】

中国期刊全文数据库 前1条

1 李毅;齐清会;;胃肠道神经-Cajal间质细胞-平滑肌网络研究进展[J];国际消化病杂志;2007年03期

本文编号：1999661