ABSTRACT
Objective To study the effect ofβ?sodium aescinate on the expressions of aquaporin?4 and aquaporin?9 in rats with spinal cord inju?ry. Methods A total of 150 rats were randomly divided into 3 groups:sham group(n=50),spinal cord injury(SCI)group(n=50)andβ?sodi?um aescinate group(n=50). The experimental animal models was established by modified Allen’s model. The Basso,Beattie and Bresnahan (BBB)locomotor rating scale and inclined plane test were used to evaluate rat behavioral consequences after injury.The immunohistochemical staining and western blotting assay were performed to observe the expressions of aquaporin?4 and aquaporin?9. Results Compared with sham group,BBB score and inclined plane test score of SCI group andβ?sodium aescinate group were significantly lower at each time point(P<0.05);however,the functional recovery was significantly better inβ?sodium aescinate group than in SCI group at each time point from 7 d after SCI(P<0.05). The aquaporin?4 and aquaporin?9 positive expressions of rats in sham group were lower significantly than rats in SCI group andβ?sodium aescinate group(P<0.05);however,the aquaporin 4 and aquaporin 9 positive expressions of rats inβ?sodium aescinate group was lower signifi?cantly than rats in SCI group at each time point(P<0.05). Conclusion β?sodium aescinate can protect the neurologic function in rats with spi?nal cord injury by decreasing aquaporin?4 and aquaporin?9 protein expression.
ABSTRACT
BACKGROUND: Bone morphogenetic protein (BMP) is a protein possesses potential activity, which can increase and enhance its activity when the bone issues are damaged, so it can be used to repair the bone defects when combined with carrier. However,there are few reports concerning it as gene therapy.OBJECTIVE: To construct recombinant retroviral vector expressing human BMP2 gene and to discuss its biological function in ostecblasts.METHODS: BMP2-specific primers were designed and synthesized according gene sequence of human BMP2 gene in Genbank, then BMP2 gena was amplified by Hifi PCR, which was recombined with cloning vector pDNR-CMV homologousiy into pDNR-CMV-BMP2 plasmid identified by BMP2 PCR and enzyme digestion of Sail and EcoRI as well as gene sequencing; recombinant plasmid pDNR-CMV-BMP2 and retroviral plasmid pLP-LNCX were recombined homologously in IoxP sites into pLP-LNCX-BMP2 plasmid transferred into packing cell line PT67 and the supernatant was collected to assay viral titre. Human osteoblast was infected with retrovirus, then the growth of cells were observed by MTT, and the expression of BMP2 protein was detected by Western blotting at 48 hours transfectionRESULTS AND CONCLUSION: Digestion, BMP2 PCR and gene sequencing of pDNR-CMV-BMP2 were coincided with expected. After transfected with plasmid pLP-LNCX-BMP2, PT67 cells could be screened with G418 only to get stably integrated in BMP2, of whose supemanant viral titra amounted to 5×10~8 pfu/mL. MTT assay showed that there had no evident difference in cellular inhibition between the normal and retrovirus groups at 72 hours after transfection (P ＞ 0.05); Western blotting showed that the BMP2 was strong expressed at 48 hours after transfection. It demonstrated that BMP2 gene was successful cloned and its retrovirus vector was constructed.