苦参素对实验性自身免疫性脑脊髓炎小鼠脊髓中P53、MDM2及IL-17表达的影响

Effect of matrine on expressions of P53,MDM2 and IL-17 in spinal cord of experimental autoimmune encephalomyelitis mice

目的:观察苦参素对实验性自身免疫性脑脊髓炎(EAE)小鼠脊髓中P53、MDM2及IL-17表达的影响。方法:将45只C57BL/6雌性小鼠随机分3组,分别为正常组、模型组、治疗组,每组15只。模型组和治疗组用MOG35-55免疫诱导建立慢性EAE模型,治疗组于免疫后第13天起腹腔注射苦参素200 mg/kg,1次/d,连用10 d。采用HE和勒克斯光蓝(LFB)染色观察3组小鼠脊髓组织病理学变化,采用免疫荧光组化染色法检测脊髓组织中P53和MDM2蛋白的表达,采用ELISA法测定脊髓匀浆中IL-17的表达。结果:与模型组相比,苦参素治疗组小鼠EAE发病时间推迟,神经功能评分降低(P<0.05),小鼠脊髓组织中炎症浸润和脱髓鞘评分降低(P<0.05),P53蛋白表达增加(P<0.01),而MDM2与IL-17的表达降低(P<0.01)。小鼠脊髓组织中P53与IL-17表达呈负相关(r=-0.665,P<0.05)。结论:苦参素可能通过调节P53/MDM2的表达,控制致炎因子的表达,从而对EAE起到一定的治疗作用。

Aim： To observe the therapeutic effect of matrine（ MAT） on experimental autoimmune encephalomyelitis（ EAE） mice,mainly focusing on the expressions of P53,MDM2 and IL-17. Methods： Forty-five female C57 BL/6 mice were randomly divided into three groups： normal group,model group and MAT group（ 15 in each group）. EAE model was induced by injecting MOG35-55 in model group and MAT group,and MAT were given from the 13 th day post immunization by intraperitoneal injection daily（ 200 mg/kg） for 10 days in the MAT group. HE staining and Luxol fast blue（ LFB） staining were carried out in spinal cord tissue in the 3 groups,and the inflammatory infiltration and demyelination were assessed. The expressions of P53 and MDM2 proteins were measured by immunofluorescence,and IL-17 was detected by ELISA. Results： Compared with those of the model group,the disease onset was delayed and clinical symptoms of EAE ameliorated（ P 0. 05）,the degree of inflammatory infiltration and demyelination was significantly inhibited in MAT group（ P 0. 01）; moreover,the expression of P53 increased（ P 0. 01）,while the levels of MDM2 and IL-17 expression were obviously reduced（ P 0. 01） in MAT group. There was a negative correlation between P53 and IL-17（ r =-0. 665,P 0. 05） in spinal cord tissue. Conclusion： Matrine has therapeutic effect on EAE by regulation the expressions of P53 and MDM2.