ABSTRACT
Objective To investigate the effects of ulinastatin(UTI)on the nerve regeneration of mice experimental allergic encephalomyelitis and the expression of related factors and protein.Methods Forty mice were randomly divided into four groups:ulinastatin group(U),atorvastatin group(A), empty control group(C)and normal control group(N).The experimental autoimmune encephalomyelitis(EAE)in mice was constructed by Freund's complete adjuvant and MOG35-55 polypeptide.Histopathological changes were observed by HE,LFB and Bielschowsky stained at the 3rd week and 4th week after immunized of each group.The expressions of CD4 +T cells were estimated by immunohistochemical method.The expression of myelin basic protein (MBP),brain-derived neurotrophic factor(BDNF),growth associated protein-43(GAP-43),2',3'-cyclic nucleotide-3'-phosphodiesterase(CNP)were detected by Western-blot.Results The largest neurological score of group U was lower than group C,and the difference was statistically significant(P<0.05 ).Pathological features showed that the inflammatory cells,demyelination of spinal cord and axonal injury of group U were lighter than group C.With the duration of treatment,nerve injury decreased.After UTI treatment,the expression of MBP,BDNF,GAP-43,CNP increased.They were statistically significant difference when compared with group C(P<0.05).There was no significant difference between ulinastatin and atorvastatin in the treatment of EAE.Conclusion Ulinastatin could reduce the extent of nerve damage effectively and promote its regeneration which provide a theoretical basis for the clinical treatment of MS.
ABSTRACT
Background and purpose: Itch protein is an established regulator of T cell immune response thresholds, belong to a class of E3 ubiquitin-transferring enzymes, widely involve in the ubiquitination of several key signaling molecules, such as ZAP70, P85, VAV, PLC-γ, PKC-θ, etc, plays a critical role in tumor induced immu-nosuppression. Itch ligase activity regulate T-cell anergy and development of regulatory T cells in the periphery by modulating key components of T-cell receptor and transforming growth factor-βsignaling. Therefore, manipulation of Itch activities may provide the opportunities to develop future therapies for immune disorders such as autoimmunity and cancer. speciifc small interfering RNA(siRNA) was utilized to silence the expression of Itch gene of T-lymphocytes and investigate the cytotoxicity activity of transfected T lymphocytes against MFC stomach neoplasms cells in vitro. Methods:T lymphocytes were isolated from the spleen of 615 mice and transfected by speciifc siRNA to silence the expression of Itch gene, The expression of Itch protein were examined by Western bolt in each group;72 hours after transfection, The secretion level of IL-2, INF-γwere measured by enzyme-linked immunosorbent assay (ELISA). At the end, the cytotoxicity activity changes against MFC stomach neoplasms cells was compared between transfected T lym-phocytes, negative control and blank control in vitro. Results:Compared with control group, the expression rate of Itch protein of transfected T-lymphocytes was decreased to 16%after transfection 48 hours;72 hours after transfection, the secretion level of IL-2 in transfection group, negative control and blank control respectively were (1 891.96±141.91)pg/mL, (1 241.69±91.67)pg/mL and (1 175.03±89.14)pg/mL (P<0.001), the secretion level of INF-γin transfection group, negative control and blank control respectively were (958.33±75.46)pg/mL, (683.33±66.67)pg/mL and (691.72±68.72) pg/mL (P<0.05). Transfected T lymphocyte also showed more efifcient killing ability against MFC stomach neoplasms cells than negative control and blank control in vitro, the highest killing rate has reached (54.18±2.96)%. Conclusion:Silencing Itch gene can signiifcantly promoted the secretion level of IL-2, INF-γof mice T lymphocyte, enhanced the cytotoxicity activity of T lymphocyte against MFC stomach neoplasms cells in vitro.