穴位刺激和跑台锻炼对模拟失重致肠黏膜损伤的修复作用

发布时间：2018-01-15 08:14

本文关键词：穴位刺激和跑台锻炼对模拟失重致肠黏膜损伤的修复作用　出处：《中国农业大学》2015年博士论文　论文类型：学位论文

【摘要】：失重/模拟失重对动物机体各系统都有一定的负面影响,如骨丢失,肌肉萎缩,心血管功能失调等。现有报道中关于失重/模拟失重对消化系统的影响方面研究比较少见。本研究应用大鼠尾部悬吊(TS)模拟失重环境,采用血清学、组织细胞病理学、免疫组化和Western Blot技术等方法,系统的观察大鼠在4周模拟失重之后,肝脏和胰腺功能,肠道结构,肠黏膜机械屏障相关蛋白以及肠黏膜免疫相关细胞和分子的变化,深入探讨模拟失重造成机体肠黏膜屏障损伤的机制。同时,采用肾俞、脾俞、三阴交穴位刺激(TS+ACU)和跑台锻炼(TS+RT)作为对抗措施,研究其对模拟失重造成的肠道黏膜屏障损伤的防护作用,以探讨穴位刺激作为一种新型的对抗失重/模拟失重的措施的可行性。研究结果如下： 1模拟失重对大鼠肝脏和胰腺结构的影响与对照组相比,4周尾部悬吊之后,TS组大鼠血清ALT和AST水平没有明显变化,而肝脏SOD活性显著下降(p0.05),MDA含量显著上升(p0.05),表明肝脏抗氧化能力降低；同时,观察到肝脏糖原的储存量显著下降(p0.05),Bcl-2及Caspase-3蛋白水平均显著高于对照组(p0.05),TUNEL染色结果表明TS组大鼠肝脏凋亡细胞数量显著增加(p0.05),说明尾部悬吊可促进大鼠肝脏的细胞凋亡。TS组血清INS和IGF-1水平显著降低(p0.05),提示胰腺功能可能出现异常；PAS染色表明胰腺中糖原含量下降,进一步证实胰腺功能异常。 2模拟失重对大鼠肠黏膜机械屏障的影响与对照组相比,TS组大鼠肠黏膜表现出一定程度的病理学改变,包括上皮细胞增生,十二指肠和空肠绒毛和隐窝显著变短(p0.05),肠绒毛宽度增加(p0.05),杯状细胞数量增加(p0.05)等；同时观察到肠黏膜屏障相关蛋白的表达量也表现出一定的变化,上皮细胞紧密连接蛋白ZO-1和Claudin1以及黏液屏障蛋白Mcu2表达量均显著降低(p0.05),促进肠黏膜机械形成的蛋白Numb表达量也显著下降(p0.05),而有负反馈调节作用的Hesl蛋白的表达量则显著增加(p0.05)。 3模拟失重对大鼠肠黏膜免疫的影响与对照组相比,TS组大鼠小肠上皮内淋巴细胞、潘氏细胞和肥大细胞数量均表现出不同程度的增加；同时对大鼠肠黏膜免疫相关分子进行检测的结果发现,SIgA、IL-1β、IL-6、NF-KB和CD23表达量显著增加(p0.05),而P物质表达显著降低(p0.05)。 4穴位刺激对模拟失重致大鼠肠黏膜损伤的防护作用经过穴位刺激后,与TS组相比,TS+ACU组大鼠肝脏SOD活性显著增高(p0.05),MDA含量显著下降(p0.05),肝脏糖原含量略有上升,Bax、Bcl-2及Caspase-3蛋白水平略有下降,肝脏凋亡细胞数量显著下降(p0.05)；血清INS水平显著升高(p0.05),胰腺糖原含量显著上升(p0.05),表明穴位刺激对模拟失重造成的肝脏和胰腺结构的损伤具有一定的防护作用；同时观察到针刺组失鼠小肠病理变化减轻,而且肠黏膜屏障相关蛋白Claudinl、Mcu2和Numb表达量显著升高(p0.05),Hesl蛋白表达量显著下降(p0.05),表明模拟失重大鼠在针刺后肠黏膜的屏障受损情况有所缓减；此外,针刺组大鼠小肠上皮内淋巴细胞、潘氏细胞和肥大细胞数量显著下降(p0.05),黏膜免疫相关分子SIgA、IL-1β、IL-6、NF-KB和CD23表达量显著下调(p0.05),而P物质表达显著增加(p0.05),这些指标均趋向于与空白对照组一致。表明穴位刺激对模拟失重引起的黏膜免疫损伤具有对抗作用。 5跑台锻炼对模拟失重致大鼠肠黏膜损伤的防护作用与TS组相比,TS+RT组大鼠肝脏SOD、 MDA以及血清ALT、AST、INS和IGF-1水平没有明显变化,而且肝脏Bax、caspase-3表达水平有所降低,但是差异不显著,肝脏凋亡细胞数量显著降低(p0.05),提示跑台锻炼对模拟失重所造成的大鼠肝脏和胰腺结构损伤的对抗作用不明显；同时观察到大鼠小肠病变较TS组减轻,包括十二指肠绒毛长度和隐窝深度显著上升(p0.05),绒毛宽度显著降低(p0.05),而且黏膜杯状细胞数量显著增加(p0.05)；此外,肠上皮细胞间紧密连接蛋白ZO-1和Claudinl以及调节蛋白Numb的表达量显著升高(p0.05),黏蛋白Muc2表达量也显著增加(p0.05),表明跑台锻炼对4周模拟失重引起的大鼠肠黏膜机械屏障的损伤具有一定的对抗作用；跑台锻炼还可以有效对抗模拟失重引起的肠黏膜免疫相关细胞和蛋白的变化,说明跑台锻炼可以有效对抗模拟失重大鼠引起的肠黏膜免疫亢进。综上所述,4周模拟失重可以引起大鼠肝脏、胰腺及肠黏膜结构及功能相关分子的表达量发生明显的变化,肠黏膜上皮连接结构及黏液屏障受损；穴位刺激可以在一定程度上明显缓减模拟失重引起的小肠病理变化,有效的减轻肠黏膜上皮连接结构及黏液屏障受损程度,从而保护肠黏膜免疫屏障,跑台锻炼对模拟失重情况下肠黏膜屏障的损伤也有一定的防护作用。穴位刺激试验方法的建立为航天防护措施的研发提供了新思路,具有重要的理论意义与实践意义。
[Abstract]:In this study , the effects of weight loss / simulated weight loss on digestive system were studied . Compared with the control group , the level of serum ALT and AST in TS group was significantly higher than that of control group ( p < 0.05 ) . The level of serum INS and Caspase - 3 in TS group was significantly higher than that in control group ( p < 0.05 ) . Compared with the control group , the intestinal mucosa of TS group showed a certain degree of pathological changes compared with the control group , including epithelial hyperplasia , duodenum and jejunum villi and crypts ( p < 0.05 ) , increased intestinal villi width ( p < 0.05 ) , increased number of cup - shaped cells ( p0.05 ) , and significant decrease in expression of protein Numb ( p < 0.05 ) . The expression of protein Numb increased significantly ( p < 0.05 ) , while the expression of Hesl protein with negative feedback regulation increased significantly ( p < 0.05 ) . Compared with the control group , the number of lymphocytes , Pan ' s cells and mast cells in the small intestinal epithelial cells of the TS group increased significantly compared with the control group . The results showed that the expression of SIGA , IL - 1尾 , IL - 6 , NF - KB and CD23 increased significantly ( P < 0.05 ) , while P - substance expression was significantly decreased ( p < 0.05 ) . Compared with TS group , there was a significant decrease in liver SOD activity ( p < 0.05 ) , the content of Bax , Bcl - 2 and Caspase - 3 decreased slightly ( P < 0.05 ) , and the expression of Bax , Bcl - 2 and Caspase - 3 decreased significantly ( P < 0.05 ) . Compared with TS group , the levels of SOD , MDA and serum ALT , AST , INS and IGF - 1 in the liver of TS + RT group were significantly decreased ( p < 0.05 ) . In conclusion , the 4 - week simulated weight loss can cause obvious changes in the expression of the structure and function - related molecules of the liver , pancreas and intestinal mucosa of the rat , and the intestinal mucosa epithelial connection structure and the mucus barrier are damaged ; the acupuncture point stimulation can obviously slow down the pathological changes of the intestinal mucosa caused by the loss of the intestinal mucosa , thereby protecting the intestinal mucosa immune barrier , and the running platform exercise has a certain protective effect on the damage of the intestinal mucosa barrier under the condition of simulating the loss of the intestinal mucosa .

【学位授予单位】：中国农业大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R85

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