参茸复方对自身免疫性脑脊髓炎小鼠DOR-β-arrestin1-Bcl-2信号通路的影响

Effect of Shenrong Formula on DOR-β-arrestin1-Bcl-2 Signaling Pathway in Mice with Experimental Autoimmune Encephalomyelitis

目的:探讨参茸复方对自身免疫性脑脊髓炎(EAE)小鼠DOR-β-arrestin1-Bcl-2信号通路的影响。方法:将60只C57 BL/6雌性小鼠分为正常对照组、模型组、醋酸泼尼松(3.9 mg/kg)组及参茸复方高(54.6 g/kg)、中(27.3 g/kg)、低(13.65 g/kg)剂量组,除正常对照组外,剩余5组均制备EAE模型,免疫7 d后灌胃给药,正常对照组和模型组均灌胃相应体积生理盐水,给药体积均为10 mL/kg,连续给药14 d。自免疫当天起,每日对小鼠进行神经功能评分;HE染色检测各组小鼠脑和脊髓组织的细胞形态及炎性反应;免疫组化和Western Blotting检测各组小鼠脑和脊髓中DOR、β-arrestin1、Bcl-2蛋白表达。结果:与模型组比较,各给药组大鼠神经功能评分显著降低(P<0.05或P<0.01)。组织病理学检查结果显示,模型组脑和脊髓组织有大量炎性细胞聚集,核固缩明显,各给药组脑和脊髓组织病理改变均有不同程度改善。免疫组化及Western Blotting结果显示,与模型组比较,除参茸复方低剂量组外各给药组脑和脊髓组织DOR蛋白表达显著升高,β-arrestin1、Bcl-2蛋白表达显著降低(P<0.05或P<0.01)。结论:参茸复方具有缓解EAE进展的作用,其机制与DOR-β-arrestin1-Bcl-2信号通路有关。

Objective:To explore the effect of Shenrong formula on DOR-β-arrestin1-Bcl-2 signaling pathway in experimental autoimmune encephalomyelitis(EAE)mice.Methods:60 C57 BL/6 female mice were divided into normal control group,model group,prednisone acetate(3.9 mg/kg)group,Shenrong formula high(54.6 g/kg),medium(27.3 g/kg)and low(13.65 g/kg)dose groups.Except for the normal control group,EAE model was prepared in the remaining 5 groups,and the mice were given by intragastric administration 7 days after immunization.The mice of normal control group and model group were given the corresponding volume of normal saline with the administration volume of 10 mL/kg for 14 consecutive days.From the day of immunization,the daily neural function score was performed.HE staining was used to detect the cell morphology and inflammatory response of brain and spinal cord.The protein expressions of DOR,β-arrestin1 and Bcl-2 in the brain and spinal cord of each group were detected by immunohistochemistry and Western Blotting.Results:Compared with model group,the neurological function scores of mice in each administration group were significantly decreased(P<0.05 or P<0.01).The histopathological examination results showed that a large number of inflammatory cells in brain and spinal cord were clustered in the model group,and the nucleopytosis was obvious,the lesions of brain and spinal cord in each administration group were improved to varying degrees.The results of immunohistochemistry and Western Blotting showed that compared with the model group,except for the Shenrong formula low dose group,the expressions of DOR protein in brain and spinal cord tissues of drug administration groups were significantly increased,while the expressions ofβ-arrestin1 and Bcl-2 proteins were significantly decreased(P<0.05 or P<0.01).Conclusion:Shenrong formula can alleviate the progression of EAE,and its mechanism may be related to the DOR-β-arrestin1-Bcl-2 signaling pathway.