益肾达络饮对EAE小鼠乙酰化组蛋白H3的影响及机制探讨

The mechanism of the changes of acetylated histone H3 in mice with EAE

目的 探讨益肾达络饮对实验性自身免疫性脑脊髓炎(EAE)小鼠乙酰化组蛋白H3的影响及机制。方法 60只C57BL/6雌性小鼠随机分成6组:空白组、模型组、中药组、激素组、MRS2179(P2Y1受体拮抗剂)组、MRS2179+中药组。分别给予不同药物干预。每日记录各组小鼠的体重及神经功能评分,运用免疫蛋白印迹法(WB)测定小鼠脑及脊髓中P2Y1的表达含量,运用WB及免疫组织化学染色法(IHC)检测小鼠脑及脊髓中乙酰化组蛋白H3的表达水平。结果 神经功能评分的变化情况:中药组、MRS2179+中药组与激素组低于模型组与MRS2179组。各组小鼠中枢神经系统(CNS)中P2Y1的表达情况:模型组高于空白组、MRS2179组、中药组、MRS2179+中药组与激素组。乙酰化组蛋白H3的表达情况:模型组高于空白组、中药组、MRS2179+中药组、MRS2179组与激素组。差异具有统计学意义。结论 益肾达络饮可通过抑制P2Y1的表达,降低乙酰化组蛋白H3的表达水平,从而降低神经功能评分,缓解EAE的疾病状态。

Objective To investigate the mechanism of the changes of Acetylated Histone H3 in mice with EAE.Methods Sixty C57BL/6 female mice were randomly divided into 6 groups.The normal control group,model group and MRS2179(P2Y1 receptor antagonist) group were given normal saline by gavage,hormone group was given prednisone acetate,Chinese medicine group and MRS2179+Chinese medicine group were given Yishendaluo decoction.The body weight and clinical scores of the mice were recorded daily,the expression of P2Y1 in the brain and spinal cord of the mice was detected by western blotting,the expression of Acetylated Histone H3 in the brain and spinal cord of the mice were detected by western blotting and immunohistochemical staining.Results Changes in clinical scores:The Chinese medicine group,MRS2179+Chinese medicine group and hormone group were lower than the model group and MRS2179 group.The expression of P2Y1 in CNS of mice in model group was higher than that in the normal control group,MRS2179 group,Chinese medicine group,MRS2179+Chinese medicine group and hormone group.Expression of acetylated histone H3:The model group was higher than that of the normal control group,the Chinese medicine group,the MRS2179+Chinese medicine group,the MRS2179 group and the hormone group.The difference is statistically significant.Conclusion By inhibiting the expression of P2Y1,the expression level of acetylated histone H3 can be reduced,thereby reducing the clinical scores and alleviating the disease state of EAE.