| Objective: To study the effect of total flavonoids of astragalus(TFA)on the spinal cord injury(SCI),and to seek the molecular mechanism of TFA accelerating the recovery of SCI by regulating Wnt/β-catenin signaling pathway,which will lay foundation for TFA in the treatment of spinal cord injury.Methods: 32 8-week-old SD rats were got into the groups of sham group(Sham),model group(SCI),total flavonoids of astragalus group(TFA),Wnt/β-catenin inhibitor group(ICG-001).The group of SCI,TFA and ICG-001 were treated with Allen’s blow method to construct a rat SCI model.After successfully constructing the model,50.0mg/(kg·d)TFA was injected intraperitoneally;after injecting TFA,5.0 mg/(kg·d)Wnt/β-catenin inhibitor was injected via tail vein in ICG-001 group.The drug was used for 14 days,which is replaced by normal saline in Sham and SCI group.Behavior tests were assessed the effect of TFA on the coordination,range and degree of rats limb mobility on 1,3,7,14 and 21 days after constructing the model.After 21 days,the animals being killed,the tissue samples were collected.Pathology,molecular biology detection and other technologies were used to carry out research.Results: 1.BBB scores showed that after modeling for 7 d,TFA group rats gained evidently higher scores than the SCI group rats(P<0.0001).SCI group was served as a reference,the rats in ICG group were clearly better and faster on the cure of SCI after modeling for 14 days(P<0.01),but worse than that in TFA group.2.For the inclined plate experiment,TFA group made significant difference with SCI group after modeling for 14 d(P<0.01).The TFA group taken as the control,the angle was smaller in ICG-001 group,but higher than SCI group.3.HE staining showed that the morphology cell and nuclei in Sham group was normal,with aligned nuclei observied.In SCI group,there were large hemorrhagic spots,necrotic foci,inflammation,and changes in nuclear morphology and size.Compared with SCI group,TFA group showed no obvious inflammation,and significantly improved cell morphology.ICG-001 group showed the nuclear morphology was changed to some extent in the spinal cord tissue.4.TUNEL staining showed that the rate of positive cell in SCI,TFA,and ICG-001 group was obviously higher than that in Sham group(P<0.0001,P<0.0001,P<0.0001),which was clearly declined in TFA and ICG-001 group compared with SCI group(P<0.001,P <0.05),the rate of positive cell in ICG-001 group was obviously higher than that in TFA group(P<0.05).5.The results of ELISA showed that the content of serum IL-1β IL-6,TNF-α were little in Sham group,which were obviously increased in SCI,TFA and ICG-001 group(P<0.05).The cytokines mentioned above were clearly drop in TFA group compared to SCI group(P<0.05),which were roughly the same between SCI and ICG-001 group(P>0.05).The cytokines mentioned above were clearly less in TFA group compared to ICG-001 group(P<0.05).6.The results of q RT-PCR indicated that the SCI group taken as control,the expression levels of Wnt-4,β-catenin and c-myc were clearly rised in TFA group(P<0.0001,P<0.0001,P<0.001),while in ICG-001 group Wnt-4 was obviously rised,(P>0.05)but the β-catenin and c-myc was obvious unchanged(P<0.05).7.The results of Western Blot showed that the SCI group being served as reference,the level of Wnt-4,β-catenin and c-myc in TFA group were clearly increased(P<0.001,P<0.0001,P<0.001),while in ICG-001 group Wnt-4 was obviously rised(P>0.05),but the β-catenin and c-myc made no significant difference in ICG-001 group(P<0.05).Conclusion: It can be concluded that total flavonoids of Astragalus can promote the nerve repair and improve the motor ability of rats with spinal cord injury,and its molecular mechanism may be related to the activation of Wnt/β-catenin signaling pathway,thereby inhibiting the expression of inflammatory factors and inhibiting cell apoptosis. |
PDF Full Text Request