The Study Of Palmitoleic Acid Improving Symptoms Of AGVHD Mouse Model

Objective:To evaluate the impact of palmitoleic acid(POA)on lymphocytes and the mechanism ofPOA’s modulating cholesterol metabolism in mice.Furthermore,to investigate the effect ofPOA in acute graft-versus-host disease(a GVHD)mice model.Methods:1.To study the immunosuppressive effect ofPOA on lymphocytes activation and the population of Regulatory cell in vivo.1.1 Using cell counting kit(CCK-8 kit)respectively to detect the immunosuppressive effects ofPOA at different concentrations on the proliferation of lymphocyte.1.2 Using flow cytometry to detect the inhibitory effect ofPOA on T lymphocytes activated by phorbol myristate acetate(PMA)and Ionomycin(IM).2.To study the Mechanism of palmitoleic acid improving hypercholesterolemia in mice.2.1 Establish a hypercholesterolemic mouse model,treat it with tPOA and cPOA respectively,than measure the blood lipid level in mice serum and liver.2.2 Western Blot and real-time polymerase chain reaction(RT-PCR)were used to detect the expression of 3-hydroxy-3-methylglutaryl coenzyme reductase(HMGCR)gene and protein in liver.2.3 Western Blot and RT-PCR were used to detect the gene and protein expression level of cholesterol 7-αhydroxylase(CYP7A1).3.To study the effects of palmitoleic acid on survival and prognosis of a GVHD mice.Establish mouse a GVHD model,treat withPOA,observe and record the clinical manifestations and survival of mice.Result:1.POA can inhibit the proliferation and modulate immunity on lymphocyte in vivo.1.1 The cPOA has no cytotoxicity to lymphocytes.The proliferation activity of lymphocytes separated from mice spleen was not affected by cPOA at the concentration of 250-500μM.1.2 The cPOA concentration of 250μM-500μM can effectively inhibit the proliferation of T lymphocytes activated by PMA and IM,and effectively increase the proportion of CD4~+CD25~+cells.1.3 The a GVHD mice had clinical manifestations such as fluffy hair,depilation,anus become red and turgescence,and loose stools.there were no a GVHD pathological changes in the liver and small intestine on the control group.In the a GVHD group,the pathological sections of the liver and the small intestine had typical changes of a GVHD characteristics such as intracellular edema,capillary congestion in the lamina propria and disordered tissue structure.1.4 The therapeutic effect of cPOA on a GVHD mice model showed that the survival time of mice in the treatment group was significantly prolonged and the clinical score of a GVHD was decreased.2.POA against hypercholesterolemia by regulating cholesterol metabolism.2.1 The tPOA can reduce serum cholesterol(TC),low density lipoprotein(LDL-c),high density lipoprotein(HDL-c),free cholesterol(FC)and total bile acid(TBA)in liver.However,except TBA,the cPOA has no statistical significance in reducing these indexes.2.2 Both tPOA and cPOA decreased the expression of HMGCR gene and protein in liver.2.3 Both tPOA and cPOA increased the expression of CYP7A1 in liver,but there was no significant difference between the group treated with low dose of cPOA(150mg/kg)and the control.3.POA significantly prolonged the survival time and improved the clinical symptoms of a GVHD mice.Conclusion:POA has immunosuppressive effect on lymphocytes in vitro and has effect on hypercholesterolemia by regulates cholesterol metabolism.It has potential application value in improving the symptoms of a GVHD.