| Objective: use of mesenchymal stem cells (Mesenchymal stem cells, MSCs) immunomodulatoryeffects and low immunogenicity characteristics investigate immunomodulatory effects of long-termkidney function after transplantation its rat kidney and brain death.Methods:(1) inbred between male F344rat bone marrow-derived mesenchymal stem cells adherentisolation, culture, purification and identification of flow cytometry, passage, cryopreservation.(2)almost inbred male F344rats (body weight200g~250g) as a donor, inbred,(body mass200g~250g)male Lewis rats as recipients. Divided into3groups:①normal control group (G1, n=5): F344ratstake the left kidney, in situ removal of the left kidney was implanted Lewis rats;②brain death group(G2, n=5): The big F344induced rat brain death, whichever is6hours later the left kidney, in situremoval of the left kidney was implanted Lewis rats;③brain death+MSCs group (G3, n=5): TheF344rats induced brain death, infusion of MSCs,6hours later to take the left kidney, and thenimplanted in the left kidney was removed in Lewis rats;(3) All Lewis rat kidney transplant for10consecutive days from the date given daily intramuscular injection of cyclosporine A (0.15mg/100gbody weight of rats), the first10days after the removal of the right kidney, after the first14days,21days,28days,35days, respectively, blood, serum creatinine (Scr);(4)35days after the transplantkidney specimens obtained and The first10days after the removal of the right kidney specimens①immunohistochemistry;②renal allograft histology.Results: In this study, adherence separation spread to the third generation of cells obtained by flowcytometry with low expression of CD34, CD45, high expression of cell characteristics CD29, CD105and CD44, consistent with the phenotypic characteristics of MSCs. These cells by light microscopyshowed fibroblast-like, spindle, more uniform shape, in addition, they can be induced to differentiateinto adipocytes, confirmed from inbred male F344rat bone marrow isolated, cultured, purified cells areMSCs. All rats with brain death are in line with established brain death donor requirements:(deepcoma, respiratory arrest, unconditional reflex, pupillary light reflex, corneal reflex, rats were observedin mean arterial pressure is still greater than6hours80mmHg). Detection of renal transplantation Scrthree groups of rats at different times to assess the function of the transplanted kidney, Scr rats than G1G2, G3group is high, and the difference was statistically significant (P <0.01), G3rats Scr slightlyhigher than the G1group, the difference was not statistically significant (P>0.05). Pathology of renal tissue from each group HE staining in the light microscope and found that G2group appeared obviousinfiltration of mononuclear cells and renal tubular epithelial inflammation (P <0.05), G3and G1groupsshowed no significant difference (P>0.05); three groups of transplanted kidney arteries havesignificant inflammatory changes and severe than G1and G3group G2group, but no significantdifference between the three groups (P>0.05). Immunohistochemical analysis showed G2grouptransplanted kidney glomeruli, tubules and interstitial cells IL-1β, TNF-α expression was positive G1expression,, G3group were lower than the G2group (P <0.05).Conclusion: MSCs can regulate brain death donor kidneys of rats immune status, reduce inflammationof kidney damage, kidney transplantation has a protective effect on the receptor, reducing thetransplanted kidney tissue IL-1β, TNF-α inflammatory cytokine production, the transplanted kidneycan get good long-term function. |
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