内镜下放置羧甲基纤维素膜对食管环周型内镜黏膜下剥离术后狭窄的预防

发布时间：2018-03-21 00:35

本文选题：内镜黏膜下剥离术(ESD)　切入点：羧甲基纤维素膜(CMC)　出处：《第二军医大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究目的:探讨羧甲基纤维素膜预防食管环周型ESD术后狭窄的有效性及相关可能机制。研究方法:本实验分为两部分。第一部分为动物实验。将14只巴马猪分为两组,分别为CMC组(n=7)和对照组(n=7)。通过术前健康观察和禁食后,在全麻状态下,于食管40-45cm段长约5cm处行食管环周ESD手术。CMC组术后立即在创面放置CMC膜,对照组创面则不予处理,两组实验动物均于术后14天处死。术后分别观察记录两组动物的体重变化、吞咽困难评分、内镜复查结果以及对处死后所获取的创面组织进行的狭窄数据的测量。第二部分为基础实验研究。对术后14天获取的食管创面组织进行组织病理学检测,通过HE染色、Masson染色对比分析两组间的固有肌层损伤、黏膜下层纤维化程度和上皮再生情况。使用Elisa方法分别检测术前0天和术后14天动物血清中TGF-β1、TNF-α、IL-1β、IL-6的含量。通过实时荧光定量PCR和Western blot分别检测术后7天食管创面愈合组织中TGF-β1、CTGF、α-sma的mRNA和蛋白表达水平。结果:第一部分结果:两组体重在术后14天均下降,但CMC组体重减少百分比(3.09±1.63)%与对照组体重减少百分比(4.97±3.40)%之间无统计学差异,p=0.220。术后7天,CMC组吞咽困难指数为(1.1±0.7)低于对照组(2.0±0.8),但差别无统计学意义,p=0.055;术后14天,CMC组吞咽困难指数为(2.3±0.8)低于对照组(3.3±0.8),差别有统计学意义,p=0.029。内镜下术后14天,两组动物食管均狭窄,但CMC组创面愈合情况好于对照组。经过大体标本测量,CMC组食管黏膜狭窄率为(53.0±5.6)%低于对照组(72.1±4.0)%,两组间有统计学意义,p㩳0.05。第二部分结果:通过组织病理学检测,CMC组新生上皮长度为(7.21±0.86)mm长于对照组(5.30±1.25)mm,差异有统计学意义,p=0.006;CMC组食管黏膜下层纤维化厚度为(1.01±0.24)mm要低于对照组(1.54±0.21)mm,差异有统计学意义,p=0.001;固有肌层损伤程度评分CMC组中位数为1(range 0-2)低于对照组中位数2(range2-3),且具有统计学意义,p=0.016。术后14天TGF-β1因子水平,CMC组(3285.71±531.61)要低于对照组(4593.71±771.03),p=0.003;且两组术前术后TGF-β1因子差值也有统计学差异,p=0.021;而另外三个因子TNF-α、IL-1β、IL-6两组间对比均无显著差异。通过实时荧光定量PCR和Western blot检测,CMC组与对照组相比,食管创面组织TGF-β1、CTGF、α-sma的mRNA和蛋白表达水平均下降,且均具有统计学差异,p0.05。结论:内镜下放置CMC膜于食管环周ESD术后创面,可以有效减轻术后猪食管狭窄进程,为预防食管环周ESD术后狭窄提供了新的策略。
[Abstract]:Objective: to investigate the effectiveness of carboxymethylcellulose membrane in preventing esophageal circumferential ESD stenosis and its possible mechanism. Methods: the experiment was divided into two parts. The first part was animal experiment. Fourteen Bama pigs were divided into two groups. CMC group (n = 7) and control group (n = 7). After health observation and fasting before operation, under general anesthesia, CMC membrane was placed on the wound in the esophageal circumferential ESD group at a length of about 5 cm between 40 cm and 45 cm after general anesthesia, while the wound in the control group was not treated. The experimental animals in both groups were killed on the 14th day after operation. The weight changes and dysphagia scores of the two groups were observed and recorded respectively after operation. The results of endoscopic reexamination and the measurement of stricture data of wound tissue obtained after death. The second part is a basic experimental study. Histopathological examination of esophageal wound tissue acquired 14 days after operation was carried out. The injury of the lamina propria muscularis between the two groups was analyzed by HE staining and Masson staining. The degree of submucosal fibrosis and epithelial regeneration were measured by Elisa method. The serum TGF- 尾 1 and TNF- 伪 TNF- 伪 IL-1 尾 -IL-6 were measured by Elisa method. The recovery of esophageal wound was detected by real-time fluorescence quantitative PCR and Western blot. The expression of mRNA and protein in TGF- 尾 1 CTGF, 伪 -sma was observed. Results: the first part: the body weight of the two groups decreased 14 days after operation. However, there was no significant difference in weight loss percentage between CMC group and control group by 3.09 卤1.63% and 4.97 卤3.40% respectively. The dysphagia index of CMC group was 1.1 卤0.7 on the 7th day after operation, which was lower than that of control group (2.0 卤0.8), but the difference was not statistically significant, and 14 days after operation, the dysphagia index of CMC group was lower than that of control group, but the difference was not statistically significant, and 14 days after operation, the dysphagia index of CMC group was lower than that of control group. The difficulty index was 2.3 卤0.8), which was significantly lower than that in the control group (3.3 卤0.8). The rate of esophageal mucosal stenosis in the CMC group was 53.0 卤5.6% lower than that in the control group (72.1 卤4.0%), and there was significant difference between the two groups. Results of the second part: the length of neonate epithelium in CMC group was 7.21 卤0.86 mm longer than that in control group (5.30 卤1.25 mm), and the thickness of submucosal fibrosis in CMC group was 1.01 卤0.24 mm lower than that in control group (1.54 卤0.21 mm). The median of myometrium injury in the CMC group was lower than that in the control group (1 range 0-2), which was significantly lower than that in the control group (P 0.016). The level of TGF- 尾 1 factor in the CMC group was 3285.71 卤531.61on the 14th day after operation, which was lower than that in the control group (4593.71 卤771.03p0.003), and the difference of TGF- 尾 1 factor between the two groups before and after operation was also lower than that in the control group. There were significant differences between the two groups, but there was no significant difference between the three other factors, TNF- 伪, IL-1 尾 and IL-6. The results of real-time fluorescence quantitative PCR and Western blot were compared with those of the control group. The expression levels of mRNA and protein of TGF- 尾 1 CTGF and 伪 -sma in esophageal wound tissue were all decreased, and the difference was statistically significant (P 0.05). Conclusion: placing CMC membrane into the wound surface of esophageal circumferential ESD under endoscope can effectively alleviate the process of esophageal stricture after operation. It provides a new strategy to prevent the stricture of periesophageal ESD.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.1

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