| ObjectiveThis study aimed to treat spinal cord injury rats with Trllium tschonoskii Maxim based on the traditional Chinese medicine method of activating blood circulation,and to observe the therapeutic effect.By detecting the molecular expression of Nrf2/ARE signaling pathway related proteins,to explore the mechanism of the protective effect of TTM on neurons.Methods150 SD male rats were randomly divided into sham group,model group,TTM low-dose group,TTM medium-dose group and TTM high-dose group,with 30 rats in each group.The model of spinal cord injury was established by modified Allen's heavy strike method in the model group and the drug group,but not in the sham operation group.The TTM intervention group was given a gavage of low(50mg/kg),medium(100mg/kg)and high(200mg/kg)everyday,at the same time,the sham group and the model group received normal saline by gavage once a day.On the 1st,3rd and7 th day after operation,Behavioral-Basso-Beattie-Bresnahan(BBB)scoring was performed;The morphological changes of hematoxylin-eosin staining method and nissl staining method were observed,and the changes of MDA and SOD of lipid peroxide were detected by elisa;and the expression of Nrf2,Keap1,NQO1 and HO1 proteins were detected by Western blot(Western Blot,WB).Results1 Postoperative behavioral scores of SD rats: BBB scores of the sham operation group were close to 21 on the 1st,3rd and 7th day after surgery.On the 1st day after surgery,there was no difference between the model group and the TTM intervention groups(p>0.05).On the 3rd day after the operation,the scores of the high dose group were higher than that of the model group(p<0.05).On the 7th day after surgery,the scores of the medium and high dose intervention groups were higher than those of the model group(p<0.05).2 Microscopic observation of spinal cord tissue: HE staining results showed thatthe spinal cord tissue structure was intact and the morphology of neurons was clear in each time period of rats in the sham operation group.In the model group,inflammatory cell infiltration,neuronal cell body shrinkage and intercellular space enlargement were observed in the spinal cord tissues.In the high dose intervention group,the spinal cord structures were intact;inflammatory cells were reduced,and the morphology of neurons was normal.Nissl staining showed that the neurons in the spinal cord of the sham operation group were intact at each time period,and the number and distribution of the intracellular nissl bodies were more and equally,showing a tiger lily appearance.In the model group,the morphology of neurons was irregular,the number of nissens bodies decreased,the distribution was uneven,and even disappeared.In the high-dose intervention group,the morphology of neurons was better and the number of intracellular nissl bodies was more and the distribution was more uniform.3 Detection of SOD activity and MDA content in spinal cord tissue of rats: Elisa results showed that compared with the sham operation group,SOD activity in the model group decreased.Compared with the model group,the activity of SOD in each trillium intervention group was significantly increased(p<0.05).MDA content in the model group was increased,and significantly decreased after the intervention of trillium.4 Expression of Nrf2/ARE signaling pathway molecules in rat's spinal cord tissues: Western blot analysis indicated that Nrf2,Keap1,HO1,and NQO1 were all increased in the spinal cord tissues of the model group during each postoperative period compared with the sham group(p<0.05).On the 1st day after surgery,the expressions of Nrf2 and Keap1 in the medium-dose and high-dose groups were higher than those in the model group(p<0.05),while the expressions of HO1 and NQO1 in the high-dose group were higher than those in the model group(p<0.05).On the 3rd day after surgery,the expressions of Nrf2,Keap1,HO1 and NQO1 in the medium and high dose groups were higher than those in the model group(p<0.05).On the 7th day after surgery,the expressions of Nrf2,Keap1,HO1 and NQO1 in the low-dose,medium-dose and high-dose groups were all higher than those in the model group(p<0.05).ConclusionTTM has certain effect on the treatment of spinal cord injury model in rats,can improve the lower limb motor function after spinal cord injury in rats,promote the neuronal survival and regeneration,the mechanism may be through the activity of SOD,decrease MDA level,raise Nrf2,Keap1,HO-1,NQO1 protein expression,activation of Nrf2 / ARE signaling pathway to inhibit oxidative stress reaction. |
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