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Effect of Subcutaneous Injection of "Ala"16-aFGF（1-29） on Slow Transit Constipation Mice

Author: HuXiaoDan
Tutor: LiXueLiang
School: Nanjing Medical University
Course: Internal Medicine
Keywords: Slow transit constipation Acidic fibroblast growth factor Enteric nervous system Apoptosis
CLC: R574.62
Type: Master's thesis
Year: 2011
Downloads: 40
Quote: 0
Read: 16-aFGF（1-29） on Slow Transit Constipation Mice">Download Dissertation

Abstract


Background: slow transit constipation (slow transit constipation, STC) is caused by the dysfunction of the gastrointestinal transmission intractable constipation. The existing studies suggest that the enteric nervous system (enteric nervous system, ENS), is closely related to the development of interstitial cells of Cajal (intestinal cell of Cajal, ICC), gastrointestinal hormones and STC. However, due to the specific etiology and pathogenesis of STC is still not entirely clear, the clinical treatment of mainly relies on the use of laxatives and the lack of the fundamental effective treatment. Acidic fibroblast growth factor (acidic fibroblast growth factor, aFGF) to promote cell growth, inhibition of apoptosis, nutrition and repair nerve function, and research reported that aFGF exist in the colon nervous system and play an important role, inferred the STC intestinal lesions may have a positive impact. [Ala] 16 -aFGF (1-29) and aFGF macromolecules have the same biological activity, and more clinical application, explore [Ala] 16 -aFGF (1-29) in mice STC to provide experimental basis for clinical seeking new treatment STC. Objective: To investigate the subcutaneous injection of [Ala] 16 -aFGF (1-29) the therapeutic effect in mice with slow transit constipation and possible mechanisms. Method: First, 60 ICR mice were randomly divided into model group and control group (n = 30), the subcutaneous injection morphine hydrochloride 2.5mg/kg/d, 45d, establish STC model control group, saline was the same treatment. Mice were observed in stool - according to the Bristol Stool Form Scale (BSFS) determination stool BSFS 1,2 comply with the symptoms of constipation, the BSFS 4,5 compared with normal; experiment 45d, six mice were randomly selected to promote carbon powder experimental intestinal propulsion rate confirmed STC model. Second, the first 45 days of the experiment, observe the stool of two groups of mice, fasted for 24 h mouse colon residual amount of manure and charcoal powder propelling the experimental intestinal propulsion rate. Immunohistochemical markers in colon tissue in NSE (neuron-specific marker) and caspase-3 (apoptotic protein) positive cells; Western blot detection of colon tissue NSE expression levels; dual immunofluorescence mark NSE and Caspase-3 analysis intestinal neuronal apoptosis. Third, the verification model after the establishment of the remainder of the model and the control group of mice in each group within the packet again, namely the experimental group 1, the experimental group, control group 1, control group 2, the experimental group and the experimental group 2, respectively, to be injected subcutaneously [Ala] 16 -aFGF (1-29) (300μg/kg 2 times per week for 8 weeks) and the same amount of dissolved solution treated equally, in order to observe the comparison [Ala] 16 -aFGF (1-29) STC therapeutic effect; control group and control group were given the drug and the same amount of dissolved solution treated equally to exclude the non-specific factors in the experiment. [Ala] 16 -aFGF (1-29) 8 w after treatment to observe the stool of the mice in each group, records were fasted for 24 h mouse colon residual amount of manure and charcoal powder Propulsion Laboratory detection of intestinal Road advance rate. Immunohistochemical markers in colon tissue NSE and Caspase-3 positive cells; Western blot detection of colon tissue expression of NSE; and double immunofluorescence markers NSE and Caspase-3, Statistics intestinal neuronal apoptosis. Results: First, the experiment first 45 days, the model mice stool (BSFS 1,2) of the control group (BSFS 4,5) was dry and hard; compared with the control group, model group mice were fasted for 24 h colon residual feces were significantly increased (P lt; 0.05), intestinal propulsion rate was significantly lower (P lt; 0.01), confirmed that the mice STC model successfully established. , The first 45 days of the experiment, laboratory test results (a), the enteric nervous elements are mainly distributed in the mouse intestinal myenteric plexus and submucosal plexus to the myenteric plexus; immunohistochemistry and Western blot the expression of NSE in the display model mice colon tissue was significantly lower than that in the control group (P lt; 0.05). (B) Immunohistochemistry showed that the two groups of mice colon myenteric plexus and submucosal plexus have Caspase-3 expression, model group of Caspase-3 expression levels increased significantly compared with the control group (P lt; 0.05); dual immunofluorescence area of ??the display model group apoptosis of intestinal neurons was significantly higher than the control group (P lt; 0.05) Three [Ala] 16 -aFGF (1-29) 8 w after treatment, experimental mice stool gradually turned into sausage-like soft stools (BSFS 4,5), compared with Experiment 2 the group (BSFS 1,2) significantly improved; compared with experimental group 2, the experimental group 1 mice were fasted for 24 h significantly reduced the amount of residual stool in the colon, intestinal propulsion rate increased significantly (P lt; 0.05) . The above test items was no significant difference between the experimental group and the control group (P gt; 0.05). [Ala] 16 -aFGF (1-29) treatment 8 w, laboratory test results (a), intestinal neurons were still the mouse intestinal myenteric plexus and submucosal plexus based; immunohistochemistry and Western blot showed that the experimental group 1 mice colon tissue expression of NSE were higher than the experimental group increased significantly (P lt; 0.05), compared with the control two sets of slightly lower but not statistically significant (P GT ; 0.05). (B), Immunohistochemistry showed that each group of mice colon myenteric plexus and submucosal plexus have Caspase-3 expression, the experimental group of Caspase-3 expression levels compared with Experiment 2 was significantly reduced (P lt; 0.05), compared with the control the two groups was no significant difference (P gt; 0.05); The double immunofluorescence apoptosis in experimental mice intestinal neurons area than the experimental group 2 was significantly lower (P lt; 0.05), and between the two control groups significant difference (P gt; 0.05). Conclusion: subcutaneous injection of an appropriate amount of morphine hydrochloride in 45 days in a row can be successfully established slow transit constipation animal models. Slow transit constipation and colon tissue of mice in the enteric nervous reduce the number of yuan; intestinal neuronal apoptosis may be one of the mechanisms of the enteric nervous system lesions. Subcutaneous injection of [Ala] 16 -aFGF (1-29) can significantly improve the slow transit constipation mice stool and intestinal motility. [Ala] 16 -aFGF (1-29) may be by inhibiting apoptosis of intestinal neurons in to improve slow transit constipation mouse intestinal neuropathy.

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CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Bowel disease > Colorectal disease > Colonic disease
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