长期联合应用小剂量FK506和氯化锂对大鼠坐骨神经端侧吻合后神经再生作用的观察
发布时间：2018-07-24 09:16
【摘要】：目的:1.探讨FK506和氯化锂促进大鼠坐骨神经损伤后再生的效果及机制;2.比较使用FK506和氯化锂在修复大鼠坐骨神经损伤的可行性及优越性;3.探讨不同剂量FK506和氯化锂对坐骨神经损伤后修复和再生的影响及功能恢复的评价,为临床治疗周围神经损伤奠定基础。方法:成年SD大鼠49只,在接受端侧吻合术后被随机分配到7个不同的组中:A组,7只大鼠均接受腹腔注射0.9%生理盐水1mg/kg/d;B组,7只大鼠均接受腹腔注射FK506 1mg/kg/d;C组,7只大鼠均接受腹腔注射FK506 0.5mg/kg/d;D组,7只大鼠均接受腹腔注射氯化锂85mg/kg/d;E组,7只大鼠均接受腹腔注射氯化锂40mg/kg/d;F组,7只大鼠均接受腹腔注射氯化锂40mg/kg/d和FK506 0.5mg/kg/d;G组,7只大鼠均接受腹腔注射氯化锂85mg/kg/d和FK506 1mg/kg/d;手术后,每天对大鼠进行腹腔内注射,共12周。术后大体观察实验大鼠手术切口、肢体肿胀、肢端溃疡形成及肢体恢复自主运动情况;分别2周、4周、6周、8周和12周进行功能学检查和分析。术后12周测量趾长伸肌湿重比值、检测神经电生理、免疫组织化学指标以观察评价神经再生和功能恢复的情况。结果:1.实验动物数量及大体观察实验选用大鼠49只,高剂量联合应用FK506和氯化锂组有2只因免疫抑制剂影响导致死亡,有47只模型进入到最终分析。所有大鼠术后切口均顺利愈合,无死亡和感染。少量大鼠见足底溃疡,均在两周内愈合。术后早期所有大鼠后肢运动笨拙,膝关节屈曲障碍。术后第l2周处死前观察,各使用药物治疗组大鼠的术侧膝关节手术后屈曲障碍得到了改善明显,针刺反应灵敏。而对照组改善较差,针刺反应较迟钝。术前的实验动物的体重之间无统计学的差异,然而应用药物治疗后实验组b组、c组、f组、g组动物体重明显低于对照组有统计学差异(p0.05),其中d组licl85mg/kg/d和e组licl40mg/kg/d与对照组0.9%生理盐水1mg/kg/d之间无统计学差异(p0.05)。(见图5)2.足印分析结果术后第2,4,6,8,10,12周时进行大鼠坐骨神经指数测量并比较:在各组中,正常步态的足印(左侧显示为后爪各趾充分伸展),而术侧(右侧则显示为后爪“足下垂”且屈曲挛缩,各趾内收)。术后12周时,6个药物治疗组sfi与对照组间比较差异有显著性意义(p0.05),但是各实验组之间sfi无明显统计学差异(p0.05)。(见表1)3.趾长伸肌肌湿重质量术后12周处死大鼠后切取趾长伸肌,在分析天平上称出肌肉的湿重并观察小腿肌肉萎缩情况,并按照如下公式计算:实验侧肌肉湿重与正常侧的百分比=实验侧肌肉重量/对侧肌肉重量。a组与各实验组间的差异在统计学上有意义(p0.05),实验组中b组和f组间差异无显著统计学意义(p0.05),但均优于其他实验组(p0.05)。(见表2)4.免疫组织化学检测术后12周,药物治疗组可见大量再生有髓神经纤维和无髓纤维混合,雪旺细胞形态正常,包绕再生轴突形成有髓纤维,大多数髓鞘结构发育良好,呈同心圆板层样结构,少数为未完全闭合的不成熟髓鞘。对照组,可以观察到再生的神经纤维稀疏,髓鞘发育不良。s100及nf染色,药物治疗组要优于对照组(p0.05),但是各个药物之间无统计学差异(p0.05)。(见表3,图3,图4)5.神经电生理检测药物治疗组大鼠的神经传导速度上均优于空白对照组,对照组的神经传导速度第6周和第12周的数据上并无好转倾向(p0.05),药物治疗组间两两比较后12周的数据上均明显优于第6周的数据(p0.05)。(见表4)结论:1.fk506和氯化锂对坐骨神经损伤的再生都具有明显的促进作用。2.长期小剂量联合应用fk506和氯化锂处理后可在短时间内降低周围神经损伤后发生的免疫排斥反应,从而迅速改善坐骨神经损伤后再生的微环境,以利于神经再生速度和质量的提高,取得了与高剂量联合用药组同样的治疗效果,却并没有发生致命的不良反应,对于临床应用免疫抑制剂有潜在的意义。
[Abstract]:Objective: 1. to investigate the effect and mechanism of FK506 and lithium chloride to promote the regeneration of sciatic nerve injury in rats; 2. compare the feasibility and superiority of using FK506 and lithium chloride in Repairing Sciatic nerve injury in rats; 3. to explore the effect of different doses of FK506 and lithium chloride on the repair and regeneration of sciatic nerve injury and the evaluation of functional recovery. Methods: 49 adult SD rats were randomly assigned to 7 different groups after receiving end to side anastomosis: group A, 7 rats received intraperitoneal injection of 0.9% physiological saline 1mg/kg/d; B group, 7 rats received intraperitoneal injection of FK506 1mg/kg/d; group C, and 7 rats received FK506 0.5mg/kg/d in abdominal cavity; D group, all rats. 7 rats were treated with intraperitoneal injection of lithium chloride 85mg/kg/d; in group E, 7 rats were injected with lithium chloride 40mg/kg/d intraperitoneally; group F, 7 rats received intraabdominal injection of lithium chloride 40mg/kg/d and FK506 0.5mg/kg/d; group G, and 7 rats were intraperitoneally injected with lithium chloride 85mg/kg/d and FK506 1mg/kg/d; after the operation, intraperitoneal injection of rats was carried out every day, After 12 weeks, the surgical incision, limb swelling, acromegaly ulcers and limb recovery were observed and the functional examination and analysis were carried out for 2 weeks, 4 weeks, 6 weeks, 8 weeks and 12 weeks respectively. The wet weight ratio of the extensor digitorum longus was measured at 12 weeks after the operation, and the neuroelectrochemistry and immunohistochemistry were measured to evaluate the nerve regeneration and the evaluation of nerve regeneration. Results: 1. the number of experimental animals and the gross observation were 49 rats. High dose combined use of FK506 and lithium chloride group were killed by immunosuppressive agents. 47 models entered the final analysis. All the rats were successfully healed after operation, without death and infection. A small amount of rats saw foot ulcers, all of which were found in the rats. After two weeks, all rats' hind limbs were clumsy and knee flexion disorders. After the operation L2 weeks after the operation, the flexion disorders were improved obviously after operation side knee joint operation in the drug treatment group, and the acupuncture reaction was sensitive. The control group was less improved and the acupuncture reaction was slow. The body of the experimental animals before the operation. There was no statistical difference between weight, but the body weight of B group, C group, F group and G group was significantly lower than that of control group (P0.05), among which there was no statistical difference between D group licl85mg/kg/d and E group licl40mg/kg/d and control group 0.9% physiological saline 1mg/kg/d (P0.05). (see figure 5) 2. foot print analysis of postoperative 2,4,6 At 8,10,12 weeks, the sciatic nerve index of rats was measured and compared: in each group, the foot prints of normal gait (the left side of the hind paw were fully extended), while the operation side (the right side was displayed as the hind paw "foot droop" and the flexion contracture, and the adduction of the toes). The difference between the SFI and the control group was significant 12 weeks after the operation (P 0.05) but there was no significant difference in SFI between the experimental groups (P0.05). (see Table 1) the muscle wet weight of the 3. toed extensor muscle after 12 weeks after the operation of the muscle wet weight of the extensor muscle of the 3. toes was taken and the muscle atrophy of the leg was observed and the muscle atrophy of the calf was observed. The difference between the lateral muscle weight and the contralateral muscle weight.A group and the experimental group was statistically significant (P0.05). There was no significant difference between the group B and the F group in the experimental group (P0.05), but it was better than the other experimental groups (P0.05). (see Table 2) 12 weeks after the 4. immunohistochemical examination, a large number of regenerated myelinated nerve fibers were found in the drug treatment group. Mixed with unmyelinated fibers, the morphology of Schwann cells is normal and surrounds the regeneration axon to form myelinated fibers. Most of the myelin sheath is well developed, with a concentric circular plate like structure and a few unfully closed immature myelin sheath. The control group can observe the sparsity of the regenerated nerve fibers, the myelin dysplasia.S100 and NF staining. The drug treatment group is superior to the control group. In the control group (P0.05), there was no statistical difference between each drug (P0.05). (see Table 3, figure 3, figure 4) 5. the nerve conduction velocity of rats in the neuroelectrophysiological test group was superior to that of the blank control group. The nerve conduction velocity of the control group had no improvement on the data of sixth weeks and twelfth weeks (P0.05), and 22 of the drug treatment groups were compared. The data on the following 12 weeks were significantly better than that of sixth weeks (P0.05). (see Table 4) conclusion: 1.fk506 and lithium chloride have an obvious promoting effect on the regeneration of sciatic nerve injury..2. long-term small dose combined with FK506 and lithium chloride treatment can reduce the immune rejection in a short time after the injury of the surrounding deity. The regenerated microenvironment after the injury of the sciatic nerve is beneficial to the improvement of the speed and quality of nerve regeneration. It has obtained the same therapeutic effect as the high dose combination group, but has no fatal adverse reaction. It is of potential significance for the clinical application of immunosuppressive agents.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.3
[Abstract]:Objective: 1. to investigate the effect and mechanism of FK506 and lithium chloride to promote the regeneration of sciatic nerve injury in rats; 2. compare the feasibility and superiority of using FK506 and lithium chloride in Repairing Sciatic nerve injury in rats; 3. to explore the effect of different doses of FK506 and lithium chloride on the repair and regeneration of sciatic nerve injury and the evaluation of functional recovery. Methods: 49 adult SD rats were randomly assigned to 7 different groups after receiving end to side anastomosis: group A, 7 rats received intraperitoneal injection of 0.9% physiological saline 1mg/kg/d; B group, 7 rats received intraperitoneal injection of FK506 1mg/kg/d; group C, and 7 rats received FK506 0.5mg/kg/d in abdominal cavity; D group, all rats. 7 rats were treated with intraperitoneal injection of lithium chloride 85mg/kg/d; in group E, 7 rats were injected with lithium chloride 40mg/kg/d intraperitoneally; group F, 7 rats received intraabdominal injection of lithium chloride 40mg/kg/d and FK506 0.5mg/kg/d; group G, and 7 rats were intraperitoneally injected with lithium chloride 85mg/kg/d and FK506 1mg/kg/d; after the operation, intraperitoneal injection of rats was carried out every day, After 12 weeks, the surgical incision, limb swelling, acromegaly ulcers and limb recovery were observed and the functional examination and analysis were carried out for 2 weeks, 4 weeks, 6 weeks, 8 weeks and 12 weeks respectively. The wet weight ratio of the extensor digitorum longus was measured at 12 weeks after the operation, and the neuroelectrochemistry and immunohistochemistry were measured to evaluate the nerve regeneration and the evaluation of nerve regeneration. Results: 1. the number of experimental animals and the gross observation were 49 rats. High dose combined use of FK506 and lithium chloride group were killed by immunosuppressive agents. 47 models entered the final analysis. All the rats were successfully healed after operation, without death and infection. A small amount of rats saw foot ulcers, all of which were found in the rats. After two weeks, all rats' hind limbs were clumsy and knee flexion disorders. After the operation L2 weeks after the operation, the flexion disorders were improved obviously after operation side knee joint operation in the drug treatment group, and the acupuncture reaction was sensitive. The control group was less improved and the acupuncture reaction was slow. The body of the experimental animals before the operation. There was no statistical difference between weight, but the body weight of B group, C group, F group and G group was significantly lower than that of control group (P0.05), among which there was no statistical difference between D group licl85mg/kg/d and E group licl40mg/kg/d and control group 0.9% physiological saline 1mg/kg/d (P0.05). (see figure 5) 2. foot print analysis of postoperative 2,4,6 At 8,10,12 weeks, the sciatic nerve index of rats was measured and compared: in each group, the foot prints of normal gait (the left side of the hind paw were fully extended), while the operation side (the right side was displayed as the hind paw "foot droop" and the flexion contracture, and the adduction of the toes). The difference between the SFI and the control group was significant 12 weeks after the operation (P 0.05) but there was no significant difference in SFI between the experimental groups (P0.05). (see Table 1) the muscle wet weight of the 3. toed extensor muscle after 12 weeks after the operation of the muscle wet weight of the extensor muscle of the 3. toes was taken and the muscle atrophy of the leg was observed and the muscle atrophy of the calf was observed. The difference between the lateral muscle weight and the contralateral muscle weight.A group and the experimental group was statistically significant (P0.05). There was no significant difference between the group B and the F group in the experimental group (P0.05), but it was better than the other experimental groups (P0.05). (see Table 2) 12 weeks after the 4. immunohistochemical examination, a large number of regenerated myelinated nerve fibers were found in the drug treatment group. Mixed with unmyelinated fibers, the morphology of Schwann cells is normal and surrounds the regeneration axon to form myelinated fibers. Most of the myelin sheath is well developed, with a concentric circular plate like structure and a few unfully closed immature myelin sheath. The control group can observe the sparsity of the regenerated nerve fibers, the myelin dysplasia.S100 and NF staining. The drug treatment group is superior to the control group. In the control group (P0.05), there was no statistical difference between each drug (P0.05). (see Table 3, figure 3, figure 4) 5. the nerve conduction velocity of rats in the neuroelectrophysiological test group was superior to that of the blank control group. The nerve conduction velocity of the control group had no improvement on the data of sixth weeks and twelfth weeks (P0.05), and 22 of the drug treatment groups were compared. The data on the following 12 weeks were significantly better than that of sixth weeks (P0.05). (see Table 4) conclusion: 1.fk506 and lithium chloride have an obvious promoting effect on the regeneration of sciatic nerve injury..2. long-term small dose combined with FK506 and lithium chloride treatment can reduce the immune rejection in a short time after the injury of the surrounding deity. The regenerated microenvironment after the injury of the sciatic nerve is beneficial to the improvement of the speed and quality of nerve regeneration. It has obtained the same therapeutic effect as the high dose combination group, but has no fatal adverse reaction. It is of potential significance for the clinical application of immunosuppressive agents.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.3
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