The Protective Role Of Morrroniside On Spinal Cord Injury In Rats And Its Mechanism

Objective: To observe the therapeutic effect of morrroniside after spinal cord injury(SCI)in rats and confirme the neuroprotective effect of morrroniside on spinal cord injury.To explore the mechanism of monoside’s protective effect on spinal cord injury.Methods:(1)128 adult female SD rats were selected,and SD rat spinal cord injury models were prepared by NYU method.(2)Basso beattie bresnahan score(BBB score)system was used to evaluate the SD rats’ motor function at 1d,3d,1w,2w,3w,4w,5w,6w after SCI.(3)6w after spinal cord injury,continuous frozen sections were prepared,and the area of injured spinal cord tissue was observed with hematoxylin-eosin staining(HE);myelin sheath retention of injured spinal cord tissue was observed with fast blue(LFB)staining.(4)Western Blot method was used to detect the expression of Caspase-3at 1d,3d,1w,and 2w after SCI;immunofluorescence double staining was used to detect oligodendrocytes(CNP)and nerves in spinal cord tissues at 1W and 2W after SCI.Number of neurons(Neu N)and apoptosis;Nissal Staining was used to observe the number of 6W spinal anterior horn motor neurons after SCI.(5)Heart perfusion samples were taken at different time points after spinal cord injury.0.01 M PBS perfusion was used to detect the concentration of oxidative stress products and antioxidant enzyme activity,and 4% PFA(0.01 M,PBS)was used to detect immunofluorescence.The length and length of the material were 0.5 cm before and after the center of the spinal cord injury.(6)Spinal cord tissue homogenate was prepared from PBS at 1d,3d,1w,and 2w after SCI.Hydrogen peroxide(H2O2)and malondialdehyde(MDA)kits were used to detect the content of H2O2 and MDA in the spinal cord tissue.Western Blot method was used to detect the content of 3-nitrotyrosine(3-NT)in spinal cord tissue;ELISA method was used to detect the content of8-hydroxydeoxyguanosine(8-OHDG)in spinal cord tissue.(7)Spinal cord tissue homogenate was prepared,and catalase(CAT),glutathionase(GSH-PX),and superoxide dismutase(SOD)kits were used to detect SCI at 1d,3d,1w,and 2w time the activity of CAT,GSH-PX,SOD in spinal cord tissues.Results:(1)Spinal cord injury model of SD rats was successfully prepared.(2)Behavioral BBB score results showed that the recovery of motor function of both hind limbs of SD rats in the mononoside administration group was higher than that of the control group at 3w,4w,5w,and 6w after SCI(P <0.05).(3)Statistical results of HE staining showed that compared with the control group,the area of spinal cord injury in the monoglycoside-administered group decreased at the injury center and at 1mm and2 mm before and after(P <0.05).(4)The results of LFB staining showed that compared with the control group,the myelin sheath retention rate in the 1mm and 2mm mononoside administration groups was increased at the injury center and before and after(P <0.05).(5)WB test found that the content of Caspase-3 protein in the injured spinal cord tissue of rats in the mononoside administration group was significantly lower than that of the control group at 3d,1w,and 2w after SCI;immunofluorescence Neu N and Caspase-3 were both The results showed that the number of Neu N-positive neurons in the injured spinal cord tissues of the rats treated with 1 and 2w mononoside after spinal cord injury was significantly higher than that of the control group(P <0.05),while the proportion of Neu N and Caspase-3 double positive cells It was significantly lower than the control group(P <0.05);similarly,the results of double staining of CNPase and Caspase-3 showed that the number of CNPase-positive oligodendrocytes in the injured spinal cord tissues of the rats treated with mononosine at 1w and 2w after spinal cord injury Significantly higher than the control group(P <0.05),and the proportion of CNPase and Caspase-3 double positive cells was significantly lower than the control group(P <0.05);Nissal staining statistics showed that compared with the control group,the distance from the injury center was 3 mm At 4mm,the number of motor horns in the anterior horn of the monoside administration group increased significantly,and there was a statistical difference(P <0.05).(6)Compared with the control group,the oxidative stress products H2O2,lipid peroxide MDA,and protein oxidation in the spinal cord injury group of the mononoside administration group at time points such as 1d,3d,1w,and 2w after spinal cord injury The content of product3-NT and DNA oxidation damage product 8-OHDG and so on both decreased(P <0.05),and reached the peak at 1d and 3d.(7)The activity of antioxidant enzymes such as CAT,GSH-PX,SOD in the injured spinal cord tissues of the rats treated with mononoside at time points 1d,3d,1w,2w after spinal cord injury were significantly higher than those in the control group(P < 0.05).Conclusion: 1.Mononoside promotes histological repair and motor function recovery of the spinal cord of rats after SCI,promotes neuronal cell survival,and has neuroprotective effects.2.Inhibiting the oxidative stress response and increasing the activity of antioxidant enzymes of the injured spinal cord tissue after SCI may be an important mechanism for the monoprotective effect of mononoside.