| Objective: The changes of β-arrestin1 and CAMP-PKA-CREB protein content in brain and spinal cord of EAE mice were measured.By the intervention of Yishen Jiedu Tongluo method,the disease degree of EAE before and after the changes of protein content was compared,and the regulation mechanism of Yishen Jiedu Tongluo method on EAE mice was comprehensively analyzed.Methods: 6-8 weeks old C57BL/6 female mice were randomly divided into 6 groups:blank group,model group,Chinese medicine group,hormone group,AAV interference group,AAV interference plus Chinese medicine group,10 mice in each group,and EAE model was established.On the 7th day after immunization,the model group,the blank group and the AAV interference group were given normal saline gavage,the hormone group was given prednisone acetate suspension by gavage,and the Chinese medicine group,AAV interference plus Chinese medicine group was given the traditional Chinese medicine compound Yishen Daluo Yin Gavage was given for 14 days,and the brain and spinal cord were taken out on the 22 nd day.Finally,Western Blot was used to detect the protein expression levels of β-arrestin1,CAMP,PKA,and CREB in the brain and spinal cord of each group of mice.Results: 1.From the weight measurement results of the mice,it was known that the mice in the blank group were in a stable condition and their weight increased slowly.The weight of the other five groups dropped sharply on the second day of immunization.The weight loss of the model group was particularly significant.According to the neurological function scores of the mice,in the early stage of the disease,the five groups of mice first developed tail tip drooping and gait changes.On the 9-15 days of modeling,the body weight of the model group mice showed a significant downward trend,and the disease was the most serious.He has had complete weakness at the base of the tail,obvious arched back,weakness or even paralysis of hind limbs or both hind limbs,intermittent claudication,loss of fur color,listlessness,and nerve function score of up to 4 points,with an average score of 3.81 points.In contrast,the mice in the Chinese medicine group did not lose weight on the 7-14 days of modeling.The symptoms were mainly tail tip drooping,gait changes,hind limbs or both hindlimb weakness,and intermittent claudication.The nerve function of the Chinese medicine group The highest score was 2.55 points and the lowest average score was 1.83 points.The mice in the AAV interference group were injected with adeno-associated virus on the seventh day of modeling,and their body weight showed a slow growth trend.At first,their neurological function score was slightly lower than that of the traditional Chinese medicine group,but after the 14 th day of immunization,the neurological function score of the AAV interference group was higher than that of the traditional Chinese medicine group.The traditional Chinese medicine group is lower than the model group.There is no significant difference between the AAV interference plus Chinese medicine group and the Chinese medicine group.There was no significant difference between the Chinese medicine group and the hormone group.2.βarrestin1 protein has the highest expression in mouse brain and spinal cord,model group> AAV interference group> Chinese medicine group,hormone group,AAV interference plus Chinese medicine group;model group mouse brain and spinal cord CAMP,PKA,CREB protein expression was lower than the other five groups.conclusion:1.The expression of 1 βarrestin1 protein is positively correlated with the pathogenesis of EAE mice,and the use of Yishen Daluo Decoction can effectively reduce the expression of βarrestin1 protein.2.Adhering to the treatment principle of "tonifying kidney,detoxifying and dredging collaterals",the method of tonifying kidney,detoxifying and dredging collaterals can increase the expression of CAMP/PKA/CREB protein in EAE mice to a certain extent,and slow down the pathogenesis of EAE characterized by demyelination mediated by CD4+T cells. |
