苏木乙醇提取物促进小鼠周围神经损伤修复的实验研究
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摘要
目的:观察苏木乙醇提取物在体内、体外实验中对Balb/c小鼠免疫机能的影响,并初步探讨其抑制免疫反应促进神经再生的作用机制。
方法:体外细胞实验取小鼠脾细胞,并在不同浓度苏木乙醇提取物及环孢素干预下观察淋巴细胞转化实验结果;体内实验观察小鼠坐骨神经损伤后应用苏木乙醇提取物对损伤局部淋巴细胞浸润及整体免疫机能的影响;坐骨神经脱髓鞘程度,有髓神经纤维再生以及神经功能恢复的情况。
结果:体外细胞实验观察到苏木乙醇提取物可以显著抑制Balb/c小鼠T/B淋巴细胞转化能力;体内实验观察到苏木乙醇提取物可以降低损伤局部及实验动物体内的细胞免疫和体液免疫强度,降低损伤及再生过程中坐骨神经脱髓鞘程度,增加有髓神经纤维再生数,促进神经功能恢复。
结论:体外细胞实验说明苏木乙醇提取物能够明显抑制Balb/c小鼠的免疫机能;体内实验说明神经损伤后将引发一系列免疫反应,苏木乙醇提取物可以降低神经损伤引发的免疫反应的强度,减少免疫反应引起的继发性神经损害,从而促进损伤神经的再生及功能恢复。
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 SME
Experimental 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.
方法:体外细胞实验取小鼠脾细胞,并在不同浓度苏木乙醇提取物及环孢素干预下观察淋巴细胞转化实验结果;体内实验观察小鼠坐骨神经损伤后应用苏木乙醇提取物对损伤局部淋巴细胞浸润及整体免疫机能的影响;坐骨神经脱髓鞘程度,有髓神经纤维再生以及神经功能恢复的情况。
结果:体外细胞实验观察到苏木乙醇提取物可以显著抑制Balb/c小鼠T/B淋巴细胞转化能力;体内实验观察到苏木乙醇提取物可以降低损伤局部及实验动物体内的细胞免疫和体液免疫强度,降低损伤及再生过程中坐骨神经脱髓鞘程度,增加有髓神经纤维再生数,促进神经功能恢复。
结论:体外细胞实验说明苏木乙醇提取物能够明显抑制Balb/c小鼠的免疫机能;体内实验说明神经损伤后将引发一系列免疫反应,苏木乙醇提取物可以降低神经损伤引发的免疫反应的强度,减少免疫反应引起的继发性神经损害,从而促进损伤神经的再生及功能恢复。
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 SME
Experimental 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.
引文
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