| Objective: The purpose of this study was to observe the effect of bone marrow mesenchymal stem cells(BMSCs)transplantation on the neurofilament expression of HD rats.Materials and Methods: 100 adult SPF male Sprague-Dawley rats were randomly divided into normal control group,stem cell control group(BMSCs),exposure group(HD),self-repair group(HD-NS)Stem cell repair group(HD-BMSCs),20 in each group.The rats in experimental group were exposed to HD solution diluted with sterile saline and injected intraperitoneally at a dose of 400 mg / kg.The control group was injected with the same dose of sterile saline.Take 1 dose daily for a duration of 5 weeks.After successful injection of the model,BMSCs were injected into the tail vein to establish a stem cell repair group.The other group was given the same dose of sterile saline to establish a self-repair group for 5 weeks.The whole picture of each group of rats recorded the general situation,record the gait score of each group.After successful establishment of the stem cell repair model,two experimental animals in each group were sacrificed by perfusion with 4% paraformaldehyde,spinal cord and ischial tissue were taken out for preparation.The rest of the animals in each group were decapitated and the spinal cord was rapidly extracted at low temperature.The ischial tissue was placed-80? refrigerator to save.The expression of NF-L,NF-M and NF-H was detected by q RT-PCR.The protein expression of NF-L,NF-M and NF-H was detected by Western Blot.Immunofluorescence staining was used to observe the expression of NF-L,NF-M and NF-H.The experimental results were statistically analyzed by SPSS17.0 statistical analysis software.Results: Control rats in the whole experiment,agile,normal development,no abnormal signs;gait score of 1,with the passage of time,the gait score remained unchanged.Give HD poisoning for 5 weeks,the rats mental state is poor,fur rough gray,thin,unable to stand,prostrate forward,after hind limbs drag,dorsiflexion,paralyzed;gait score is high,is a serious abnormal gait,And with the extension of time,always maintain a high level.After 5 weeks of transplanted BMSCs into rats exposed to HD,the body weight of rats increased significantly,the number of bilateral toe was slightly decreased when walking,and the state of paralysis was improved obviously.The score of gait decreased significantly at week 2 after BMSCs injection,And with the extension of time,the gait score decreased more obviously.Immunofluorescence staining showed that the NF-L immunofluorescence intensity of HD group was lower than that of Control group,the fluorescence intensity analysis showed that the difference was statistically significant(P<0.05);the immunofluorescence intensity of NF-L in HD-BMSCs group was higher than HD group,fluorescence intensity analysis showed that the difference was statistically significant(P<0.05).The protein expression of NF-L in HD group was significantly lower than that in Control group(P<0.05).The expression of NF-L protein in HD-BMSCs group was significantly higher than that in HD group(P<0.05).The results of PCR showed that the gene expression of NF-L in HD group was lower than that in Control group(P<0.05),and the expression of NF-L gene in HD-BMSCs group was higher than that in HD group and HD-NS group Significance(P<0.05).Gene expression is consistent with changes in protein expression.The immunofluorescence staining showed that the immunofluorescence intensity of NF-M in HD group was lower than that in Control group,and the fluorescence intensity analysis showed that the difference was statistically significant(P<0.05);the immunofluorescence intensity of NF-M in HD-BMSCs group was higher than HD group,fluorescence intensity analysis showed that the difference was statistically significant(P<0.05).The protein expression of NF-M in HD group was significantly lower than that in Control group(P <0.05).The protein expression of NF-M in HD-BMSCs group was significantly higher than that in HD group(P<0.05).PCR results showed that the gene expression of NF-M in HD group was lower than that in Control group(P<0.05),and the expression of NF-M in HD-BMSCs group was higher than that in HD group and HD-NS group Significance(P<0.05).Gene expression is consistent with changes in protein expression.The immunofluorescence staining showed that the immunofluorescence intensity of NF-H in HD group was lower than that in Control group,and the fluorescence intensity analysis showed that the difference was statistically significant(P<0.05);The immunofluorescence intensity of NF-H in HD-BMSCs group was higher than HD group,fluorescence intensity analysis showed that the difference was statistically significant(P<0.05).The protein expression of NF-H in HD group was significantly lower than that in Control group(P<0.05).The protein expression of NF-H in HD-BMSCs group was significantly higher than that in HD group(P<0.05).The results of PCR showed that the gene expression of NF-H in HD group was lower than that in Control group(P<0.05),and the expression of NF-H in HD-BMSCs group was higher than that in HD group and HD-NS group Significance(P<0.05).Gene expression is consistent with changes in protein expression.Conclusion: BMSCs transplantation for the repair of nerve damage in HD poisoning rats.Transplantation of BMSCs can up-regulate the expression of NF-L,NF-M and NF-H in the spinal cord of HD rats.BMSCs transplantation can up-regulate the expression of NF-L,NF-M and NF-H in the sciatic nerve of HD rats. |
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