| BackgroundIntestinal dysfunction can bring the whole body in a relatively severe state of malnutrition. Current available therapy includes:parenteral nutrition capable of improving the nutritional status of patients and maintaining the intestinal barrier function. However long-term parenteral nutrition can lead to liver injury and other serious complications. the intestinal transplantation is limited by high failure rate and cost.Bone marrow mesenchymal stem cells(BMSCs) are considered as multipotent progenitor cells of mesodermal origin due to their potential of self-renewal and differentiation.When body was injuried, the mobilized BMSCs would move to the injury site, and differentiate into specific tissue cells so as to being involved in self-repair under the local micro-environment. BMSCs are characterized by of low immunogenicity, drawing convenience, ease of culture, and rapid proliferation. Based on these properties, BMSCs are regarded as promising cell therapy in regenerative medicine. BMSCs can be employed in cell therapy and gene therapy, they have been widely used in cardiovascular, nervous, respiratory and trauma disorders. Clinical application has conducted under certain condition. However, we known little about how BMSCs was involved the repair of intestinal injury and fulfill their functions. The study was designed to use BMSCs in the rat model of intestinal dysfunction in vivo to observe their colonization in the intestine and their therapeutic effect.Objective1. To explore the methods of rat BMSCs isolation, culture, characterization and staining.2. Base on our previous studies, the open abdominal injury and seawater immersion rat model was employed, and pathophyiologiacal indices was characterized to conform the existence of intestinal dysfunction.3. By using fluorescent labeling and genetic markers, the colonization of male rat BMSCs in female rats'injuried intestinal tract was detected and the content of malondialdehyde (MDA) and superoxide dismutase (SOD) were investigated. Methods1. BMSCs were isolated, cultured, characterized, stained.2. The intestinal contents of MDA and SOD were investigated in the female rats model with open abdominal injury and seawater immersion. The intestinal sample obtained by biopsy were stained by HE.3. BMSCs from male rats labeled by CM-DiI were transfusion into abdominal injuried female rats through the tail vein after injury. The control group accepted saline transfusion.4. After treatment, sample from jejunum was obtained in 3,7,14,28,56 days respectively, and sectioned to detect the colonization of BMSCs in intestine by fluorescence microscopy. On the condition of lavage of circulation system,RT-PCR was applied to detect the positivity of sex determination region Y gene (SRY). Also, the content of MDA and SOD in the intestinal tissue was also detected.Results1. Compared with normal controls, the content of MDA in intestinal tract of the open abdominal injury and seawater immersion rat increased progressively from 1 hour to 3days, but the content of SOD decreased dramatically. The difference was reduced gradually and completely diminished after 14 days.2. The results of RT-PCR revealed that, the positivity of the SRY in the intestinal tissue of female rats was 50%,66.6%,33.3%,16.6%,16.6% in 3,7,14,28,56 days respectively.3. The contents of MDA and SOD in BMSCs treated rats had statistical significance in 3 and 7 days in comparison with controls.Conclusion1. The open abdominal injury with seawater immersion can lead to intestinal dysfunction in rats. MDA, SOD are better indicators of oxidative damage and antioxidative repair, and are able to reflect the intestinal damage of oxygen free radicals and antioxidant repair.2. BMSCs from male rats can colonize in the intestine injured female rat models. But the survival of cells is decreased in the long run.3.The BMSCs transplantation can promote the recovery of intestinal function. The beneficial effects of MSC are primarily mediated via paracrine actions and not by their differentiation into target cells.
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