摘要
目的研究益肾达络饮抑制β-制动蛋白1(β-arrestin1)表达对实验性自身免疫性脑脊髓炎(EAE)模型小鼠炎症因子、环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/cAMP反应元件结合蛋白(CREB)信号通路的影响,探讨益肾达络饮治疗EAE的机制。方法采用随机数字表法将60只小鼠分为正常组、模型组、中药组(益肾达络饮20 g/kg)、阳性对照组(醋酸泼尼松3.9 mg/kg)、β-arrestin1 siRNA腺相关病毒(AAV-β)组、AAV-β+中药组,每组10只。除正常组外,其余各组小鼠均制备EAE模型。AAV-β组、AAV-β+中药组小鼠通过尾静脉注射AAV-β干扰β-arrestin1蛋白的表达。造模第8天开始每天灌胃相应药液/生理盐水1次,连续14 d。检测小鼠神经功能评分,观察小鼠脑和脊髓组织的病理形态学变化,检测小鼠血清中炎症因子[白细胞介素2(IL-2)、IL-23、γ干扰素(IFN-γ)]水平,检测小鼠脑和脊髓组织中β-arrestin1、cAMP、PKA、CREB蛋白表达水平。结果与正常组比较,模型组小鼠神经功能评分、血清中炎症因子水平以及脑、脊髓组织中β-arrestin1蛋白表达水平均显著升高(P<0.05或P<0.01);脑和脊髓组织中PKA、CREB、cAMP蛋白表达水平均显著降低(P<0.05或P<0.01);脑和脊髓组织中大部分神经元可见胞体皱缩深染及炎症细胞聚集现象,且存在不同程度的脱髓鞘改变。与模型组比较,各给药组小鼠神经功能评分、脑和脊髓组织的病理形态学变化及大部分指标(除AAV-β组脑组织中的CREB、cAMP蛋白外)均显著逆转(P<0.05或P<0.01)。与AAV-β组比较,AAV-β+中药组小鼠神经功能评分、血清中IFN-γ水平、脊髓中β-arrestin1蛋白表达水平均显著降低(P<0.05或P<0.01),脑与脊髓组织中PKA、cAMP蛋白表达水平显著升高(P<0.05或P<0.01)。结论益肾达络饮可能是通过抑制中枢神经系统中β-arrestin1的表达,进而激活cAMP/PKA/CREB信号通路,减轻神经系统炎症,最终缓解EAE的症状。
OBJECTIVE To study the effects of Yishen daluo decoction on inflammatory factors and cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response element binding protein(CREB)signal pathway in experimental autoimmune encephalomyelitis(EAE)model mice by inhibiting the expressions of β-arrestin1,and to explore the mechanism of Yishen daluo decoction in the treatment of EAE.METHODS Sixty mice were randomly divided into normal group,model group,TCM group(Yishen daluo decoction 20 g/kg),positive control group(prednisone acetate 3.9 mg/kg),β-arrestin1 siRNA adenoassociated virus(AAV-β)group,AAV-β+TCM group,with 10 mice in each group.Except for normal group,EAE model was made in other groups.AAV-β group and AAV-β+TCM group were injected with AAV-βvia tail vein to interfere with the expression of β-arrestin1 protein.Starting from the 8th day of modeling,they were given corresponding drug solution/normal saline intragastrically,once a day,for consecutive 14 days.The neurological function score of mice was detected;the pathological and morphological changes were observed in the brain and spinal cord tissues of mice;the serum levels of inflammatory factors[interleukin-2(IL-2),IL-23,interferon-γ(IFN-γ)]in mice were determined;the expressions of β-arrestin1,cAMP,PKA and CREB in brain and spinal cord were detected.RESULTS Compared with normal group,neurological function scores,serum levels of inflammatory factors,and protein expressions of β-arrestin1 in brain and spinal cord were significantly increased(P<0.05 or P<0.01);protein expressions of PKA,CREB and cAMP in brain and spinal cord were decreased significantly(P<0.05 or P<0.01).The deep staining of cellular shrinkage and aggregation of inflammatory cells were observed in most neurons of the brain and spinal cord,with varying degrees of demyelinating.Compared with model group,the neurological function scores,pathological changes in brain and spinal cord tissues,and most indicators(except for CREB and cAMP proteins in the brain tissue of AAV-β group
作者
王响
王诗洋
吴元
丁文婧
谢名宗
何静文
邢天野
尚晓玲
WANG Xiang;WANG Shiyang;WU Yuan;DING Wenjing;XIE Mingzong;HE Jingwen;XING Tianye;SHANG Xiaoling(School of Basic Medicine,Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《中国药房》
CAS
北大核心
2023年第11期1326-1331,共6页
China Pharmacy
基金
国家自然科学基金资助项目(No.81873216)。
