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An Experimental Study On Enhancing Peripheral Nerve Regeneration Of Caesalpinia Sappan Ethanol Extract In Mice

Posted on:2011-11-07Degree:DoctorType:Dissertation
Country:ChinaCandidate:H ZhangFull Text:PDF
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Objective:To observe the effect of Caesalpinia sappan ethanol extract (Su Mu ethanol extract SME) on immune function, and to explore its suppression of autoimmune response to the promotion of nerve regeneration in Balb / c mice.Methods and Results:Experiments point in vitro and in vivo , in vitro experiments in different concentrations of SME intervention were observed by Balb / c mice lymphocyte proliferation and transformation function of the change; in vivo observation of different concentrations of SME after sciatic nerve injury in mice include local infiltration of lymphocytes, the change of immune function,changes in the extent of nerve demyelination, myelinated nerve fiber regeneration and recovery of neurological function, and the data for image analysis and statistical analysis.1 The study on Balb / c mice immune function inhibition of SME Experimental Methods: A total of 60 mice with sciatic nerve injury model were randomly divided into SME of high-dose group, middle dose group, low-dose group and blank control group.SME respectively 16mg/kg/d, 8mg/kg/d, 4mg/kg/d, dissolved in normal saline were fed to the control group fed the same volume of saline. Continuous drug delivery to the death, isolated and cultured spleen lymphocytes, MTT determination of splenic T / B lymphocyte transformation capability. Isolation and culture of normal Balb / c mouse spleen lymphocytes, at different concentrations of SME and cyclosporine A intervention, MTT determination of splenic T / B lymphocyte transformation capability. ELISA, after each time the concentration of circulating immune complexes in mice. Nerve samples were CD3, CD19 immunohistochemical staining.The results show that:the intervention of SME, whether in vitro or in vivo applications, can significantly inhibit lymphocyte proliferation and transformation capabilities , and its effect was dose-dependent. Serum circulating immune complexes results showed that SME can be applied to animal's overall immune function plays a certain inhibitory effect, its effect was dose-dependent. CD3, CD19 immunohistochemistry showed that ,within 2 weeks, injury department occurs lymphocyte infiltration, infiltration was negatively correlated with the dose.2 The inhibition of mice after sciatic nerve injury in autoimmune response and the mechanisms to promote nerve regeneration of SMEExperimental Methods: 84 model with sciatic nerve injury,according to the first part of the experimental design and to drug intervention groups. Observe the general state of mice in each group, ELISA determination of serum and sciatic nerve concentrations of MBP . HE stain and toluidine blue staining observed under the microscope the number of myelinated nerve fibers and myelin circumstances, skeletal muscle fibers in general morphology, cross-sectional area, fiber number and so on.Sciatic functional index and nerve conduction velocity and amplitude measurement.Experimental results show that: the application of SME without increasing the mortality rate of laboratory animals, ELISA method for measuring serum and the sciatic nerve in the concentration of MBP was negatively correlated with the dose, indicating that after sciatic nerve injury SME reduces the degree of demyelination and secondary nerve damage caused by autoimmune response after Waller degeneration. Toluidine blue staining was observed in the experimental group the number of myelinated nerve fibers was more and more uniform distribution,myelin degeneration rare was lower, whereas the control group, the small number of myelinated nerve fibers, myelin sheath degeneration more common. HE staining was observed in the experimental group more closely arranged muscle cells, muscle fiber cross-sectional area larger, a rare connective tissue proliferation; control group, muscle fiber cross-sectional area is small, muscle fiber gap widened, more connective tissue hyperplasia. Sciatic functional index and nerve conduction velocity and amplitude showed that recovery of neurological function in experimental group is better than control group.Conclusion:SME may significantly inhibit the immune function of Balb / c mice, inhibit lymphocyte proliferation transformation, reduce the degree of cell-mediated immunity and humoral immunity and within a certain range in a dose-dependent relationship.The intervention of SME after sciatic nerve injury may reduce the secondary nerve damage caused by autoimmune response after Waller degeneration and the degree of demyelination. Thereby contributing to damage nerve regeneration and functional recovery.
Keywords/Search Tags:peripheral nerve, immunity, Su Mu ethanol extract, lymphocyte, myelin basic protein, circulating immune complexes
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