FK506对异体神经移植的促神经再生作用研究进展

因创伤、肿瘤等各种原因造成较长的周围神经损坏在临床中常见,临床修复较困难,目前自体神经移植修复是首选方法,亦成为衡量各种神经桥接材料的金标准[1]。但自体神经移植存在其来源有限性,长度和直径的限制,及增加供区手术创伤和功能障碍等缺点,因此,国内外诸多学者就寻找     

绿茶多酚保存同种异体神经移植体的实验研究

[目的]研究绿茶多酚溶液保存的同种异体神经修复大鼠坐骨神经缺损的效果.[方法] 48只成年雄性Wista大鼠,随机分为4组,每组12只,将坐骨神经在梨状肌孔下5 mm 处切除1.0 cm, 神经缺损分别用4种移植物桥接.A组:自体神经移植;B组:新鲜异体神经移植;C组:经冷冻处理的异体神经移植;D组:用绿茶多酚保存的异体神经移植.术后6、12周通过大体观察、电生理学检查、组织学观察、透射电镜观察、图像分析与定量学检测评价各组修复神经缺损的效果.[结果]A、D组间差异无统计学意义(P＞0.05),A组、D组的各项指标均优于B组、C组(P＜0.05或P＜0.01).[结论]绿茶多酚溶液保存的同种异体神经是良好的神经移植替代材料.     

同种异体气管材料保存及移植实验研究

目的 探讨同种异体气管材料低温保存方法，效果并经移植验证。方法 20条实验成年犬，麻醉后，不进行气管插管，切取颈段气管6cm，再均分为实验组和对照组，实验组分别进行4℃EC液保存，慢速降温保存（每分钟1℃，5℃，10℃，20℃）和玻璃化降温保存处理，用光镜和电镜进行检查，同时，还对抗冻剂的种类，浓度和降温速率进行了筛选，另用慢速降温保存和玻璃化保存组各10段进行异体气管移植，结果 经10%DMSO（二甲基亚砜）为抗冻剂，降温速度为每分钟1℃和玻璃化法保存的气管样品组织结构完整，并仍具有组织细胞活性，为了证实保存的效果，将经低温保存的气管进行了同种异体移植，其结果较为满意，结论 该方法保存气管制 方法可行，并可应用于临床。     

犬化学去细胞神经同种异体移植的早期观察

目的：以化学去细胞同种异体神经，移植修复犬粗大神经的长段缺损，观察早期功能恢复及神经再生。方法：去细胞神经移植组和自体神经移植组各3犬，分别桥接坐骨神经5.0cm缺损。术后3个月观察其运动功能及神经再生。结果：实验组和对照组在术后功能恢复；移植段内新生神经纤维、新生血管及许旺细胞；吻合口远端有髓神经纤维等方面非常相似。结论：化学去细胞神经作为同种异体神经移植物，在修复粗大和长段神经缺损时不会被宿主排斥和吸收，其早期功能恢复及神经再生与自体神经移植无明显差别。     

FK506及神经干细胞对异体神经移植生长的作用

目的 探讨大鼠坐骨神经异体神经移植后联合应用FK506(他克莫司)及神经干细胞(NSCs)的可行性.方法 制造大鼠坐骨神经缺损模型,异体神经移植修复;根据分组不施加干预因素、吻合口注入NSCs、腹腔注射FK506,通过运动及感觉评分、电生理检测、组织学检查,以及免疫组织化学染色,观察神经再生差异.结果 同时应用FK506及NSCs组,感觉及运动评分(5.32±1.43),神经传导速度(20.52±1.45)m/s,GAP43染色(965.72±369.17)都明显优于其他组(P＜0.05).结论 FK506结合NSCs对异体神经移植生长具有促进作用.

Abstract：

Objective This experiment study the effect of FK506 and neural stem cells on the regeneration in nerve allografts. Methods An established rat sciatic nerve model was used. Sciatic nerve ( 10 mm) deficits were created in the Lewis rats. Nerves allografts were harvested from BN rats to repair the deficits of the Lewis rats. Group A received no treatment. Group B received neural stem cells. Group D received FK506. Group C receive both neural stem cells and FK506. Nerve regeneration was evaluated by standardized pin-prick and toe-spread tests. Nerve samples and gastrocnemius muscles were harvested for examination. Muscle denervation atrophy was evaluated by gastrocnemius weights. GAP-43 and BrdU (Two-step immunohistochemistry detection reagent) were assayed by immunohistochemistry test. Results Improved functional outcomes were observed in group C when compared with other groups. Nerve conduction velocity were (20. 52 + 1.45) m/s. The score of sense and motion were higher ( 5.32 ± 1.43 ) than other groups. Improved histomorphology and electrophysiologiy were observed in group C when compared with other groups. The concentrations of GAP-43 (965.72 + 369. 17) were largest in group C, showing ststistically significant difference between groups (P ＜ 0. 05). Conclusion The combination of neural stem cells and FK506 can improve nerve regeneration. It may provide an expanded source for those afflicted with extensive nerve defects.     

异体肢体移植应用 FK506 免疫抑制治疗的临床观察

目的探讨以“小剂量”FK506为基础的联合免疫抑制治疗方案在异体肢体移植中的可行性和疗效。方法2例异体肢体移植术后应用以FK506为基础的联合免疫抑制治疗方案,即口服FK5061mg,1次/d;骁悉500mg,2次/d;强的松80mg,1次/d,以后每周减5～10mg,至10mg/d维持量。结果经术后一年临床观察,移植肢体未出现明显排斥反应和常见的毒副作用,移植左手、右拇指血循环、皮肤温度、颜色与正常对侧相同,感觉恢复到指尖,功能恢复满意。结论FK506是一种安全高效的免疫抑制剂,经初步临床观察,以“小剂量”FK506为基础的联合免疫抑制治疗方案在异体肢体移植中是可行的,且毒副作用少。     

大鼠胚胎脑组织冰冻保存与同种异体移植

大鼠胚胎脑组织冰冻保存与同种异体移植杨利孙，吴声伶，易声禹，暴连喜我们用台盼蓝染色方法观察大鼠胎脑组织在液氮中保存的细胞存活率，然后将保存的组织及新鲜组织植入同种大鼠脑内，进行组织学对比研究，现报告如下。材料与方法一、胎脑组织取材正常妊娠１５～２１天...     

FK506缓释膜应用于同种异体神经移植的实验研究

目的研究普乐可复(prograf,FK506)缓释膜在同种异体神经移植中的作用。方法应用异体神经桥接大鼠坐骨神经缺损,术中局部应用FK506缓释膜,分别于术后4、8、12周对移植物行大体和光镜观察,及轴突图像分析、移植神经电镜检查、小腿三头肌肌湿重比较、患肢坐骨神经电生理检查。结果C组(应用FK506缓释膜)的神经生长最好,基本与D组(自体神经移植)相同,B组(经预处理异体神经移植)次之,A组(新鲜异体神经移植)最差。经过对肌电图、轴突计数、肌湿重的统计学分析,C、B、A组的差异有统计学意义(P<0.05),C、D组之间差异无统计学意义。结论应用FK506缓释膜有助于减轻同种异体神经的免疫排斥反应,为神经再生创造良好条件;在同种异体神经移植中应用FK506缓释膜有助于促进神经再生。     

FK506和RS-61443对大鼠异体肢体移植的联合免疫抑制作用

目的　通过大鼠异体肢体移植模型 ,旨在分析 FK5 0 6和 RS- 6 14 4 3对大鼠异体肢体移植中急性排斥反应的免疫抑制作用。　方法　选择雄性 Wistar和 SD大鼠为供、受体 ,以 FK5 0 6和 RS- 6 14 4 3为免疫抑制剂 ,对照组为术后不用药组 ,实验组根据用药剂量和药物不同分为 6组 ,各组用药时间均为 5周 (每日 1次共 2周 ,然后每周 2次共 3周 ) ,进行了 10 1例异体肢体移植动物实验。观察大鼠一般情况、移植肢体排斥反应及存活时间。　结果　对照组肢体平均存活时间为 (7.0 0± 0 .78)天 ;实验组 1～ 6组移植肢体平均存活时间分别为 (17.0 8± 4 .5 0、2 3.2 0± 5 .0 5、11.19±2 .2 8、16 .33± 1.83、13.33± 3.2 2和 5 8.76± 6 .81)天。　结论　 FK5 0 6和 RS- 6 14 4 3能抑制大鼠同种异体肢体移植术后急性移植排斥反应的发生 ,并能延长移植肢体的存活时间。     

免疫抑制剂FK506加速周围神经损伤修复后功能恢复的初步临床报告

目的探讨免疫抑制剂FK506加速临床周围神经损伤修复后功能恢复的可能性.方法严格选择15例肢体神经断裂修复术后1d～3个月志愿者,男性12例,女性3例,年龄18～45岁.口服FK506胶囊,1mg～8mg/d,定期监测血药浓度并以此调节剂量,实际用药时间2～6月.定期评价神经再生及功能恢复.结果15例患者均有效,Tinel征、肌电图、临床神经功能恢复评价等综合显示神经再生速度达2.2～4.0 mm/d,平均3.1mm/d,快于同期肢体神经断裂修复术后非免疫抑制对照组1.2mm/d的速度.FK506血药浓度维持在3～5ng/ml,所有患者未出现明显的药物副作用.结论免疫抑制剂FK506可用于周围神经损伤修复后的临床治疗,促进神经快速再生,加速肢体功能恢复,值得进一步探讨.     

FK506 in pediatric kidney transplantation—Primary and rescue experience

Between 14 December 1989 and 17 December 1993, 43 patients undergoing kidney transplantation alone at the Children's Hospital of Pittsburgh received FK506 as the primary immunosuppressive agent. The mean recipient age was 10.2±4.8 years (range 0.7–17.4 years), with 7 (16%) children under 5 years of age and 2 (5%) under 2 years of age. Fifteen (35%) children underwent retransplantation, and 5 (12%) had a panel-reactive antibody level greater than 40%. Twenty-two (51%) transplants were with cadaveric donors and 21 (49%) were with living donors. The mean follow-up was 25±14 months; there were no deaths; 1- and 3-year actuarial graft survival was 98% and 85%. The mean serum creatinine and blood urea nitrogen were 1.2±0.6 mg/dl and 26±11 mg/dl; the calculated creatinine clearance was 75±23 ml/min per 1.73 m². Twenty-four (62%) patients have been successfully with-drawn from steroids and 24 (62%) require no anti-hypertensive medication. Improved growth was seen, particularly in pre-adolescent children off steroids. Between 28 July 1990 and 2 December 1993, 24 children were referred for rescue therapy with FK506, 14.6±16.4 months (range 1.1–53.2 months) after transplantation. Nineteen (79%) were referred because of resistant rejection; 4 (17%) were referred because of proteinuria; 1 (4%) was switched because of steroid-related obesity. There were no deaths; 1-and 2-year graft survival was 75% and 68%; 17 (71%) patients were successfully rescued, including 1 of 2 patients who arrived on dialysis; 4 (24%) of the successfully rescued patients were weaned off steroids. While not without side effects, which include nephrotoxicity, neurotoxicity, diabetogenicity, and viral complications, FK506 appears to be an effective immunosuppressive agent for both primary and rescue therapy after kidney transplantation. Its steroid-sparing qualities may be of particular importance in the pediatric population.     

The effects of cold preservation and subimmunosuppressive doses of FK506 on axonal regeneration in murine peripheral nerve isografts

BACKGROUND:

FK506 is a frequently used immunosuppressant with neuroregenerative effects. The neuroregenerative and immunosuppressive mechanisms of FK506, however, are distinct, suggesting that FK506 may stimulate nerve regeneration at lower doses than are needed to induce immunosuppression. The effects of cold preservation, a technique known to improve axonal regeneration through nerve allografts, are not well studied in nerve isografts and are also reported here.

OBJECTIVES:

To determine the effects of subimmunosuppressive doses of FK506 and cold preservation on nerve regeneration in isografts.

METHODS:

The neuroregenerative properties of immunosuppressive and subimmunosuppressive doses of FK506 were compared in a murine model receiving either fresh or cold preserved nerve isografts. Sixty female BALB/cJ mice were randomized into six groups. Animals in groups I, III and V received fresh nerve isografts. Animals in groups II, IV and VI received cold-preserved nerve isografts. Mice in groups I and II received no medical therapy, while those in groups III and IV received subimmunosuppressive doses of FK506, and those in groups V and VI received immunosuppressive doses as confirmed by mixed lymphocyte reactivity assays. Nerve regeneration was evaluated with histomorphometry and functional recovery was evaluated with walking track analysis.

RESULTS:

Pretreatment with cold preservation did not significantly affect neural regeneration. The potent neuroregenerative effect of immunosuppressive doses of FK506 was confirmed, and the ability of subimmunosuppressive doses of FK506 to stimulate axonal regeneration in murine nerve isografts is reported.

CONCLUSIONS:

Less toxic subimmunosuppressive doses of FK506 retaining some neuroregenerative properties may have a clinical role in treating extensive nerve injuries.     

FK506对大鼠坐骨神经再生作用的实验研究

押目的对FK506促神经再生的作用及药物应用方法进行初步探讨。方法雄性SD大鼠60只,随机分成三组,切断双侧坐骨神经制成坐骨神经再生室模型。A组(对照组)再生室内注入生理盐水;B组(全身用药组)再生室内注入盐水,颈后皮下注射FK5061mg/kg,连续14天;C组(局部用药组)再生室内注入1μg/mlFK506。于术后一定时间内观察神经损伤局部的免疫反应熏检测腓肠肌湿重、组织形态学及图像分析、电生理学。结果B、C组神经损伤局部淋巴细胞浸润程度较A组轻微。6周时B组各测定结果明显优于A组;C组各指标好于A组,但不具有统计学意义。结论穴1雪全身应用FK506(1mg/kg)具有神经保护和神经营养作用,可加快神经功能的恢复。(2)局部应用FK506(1μg/ml)对早期损伤神经有一定的保护作用,但对神经再生的促进作用不确切。     

FK506和抗淋巴细胞血清诱导兔异体膝关节移植耐受的实验研究

目的　探讨短期使用FK5 0 6和抗淋巴细胞血清 (ALS)诱导兔同种异体膝关节移植嵌合耐受的作用。　方法　雄性大耳白兔作为供体 ,雌性新西兰白兔作为受体 ,施行同种异体异位膝关节移植 ,设单独使用ALS( 1ml/kg)组、单独使用FK5 0 6( 0 5mg/kg)组、联合使用FK5 0 6和ALS组及对照组。FK5 0 6和ALS在移植术前 1d开始使用 ,ALS连续使用 3d ,FK5 0 6连续使用 15d。观察移植物的存活时间 ,检测受体内供体血细胞嵌合率 ,进行组织切片检查。　结果　对照组平均存活 ( 13 6± 0 8)d ,单独使用ALS组为 ( 17 4± 1 8)d ,单独使用FK5 0 6组为 ( 2 3 8± 1 5 )d ,联合使用FK5 0 6和ALS组为 ( 4 0 4±2 9)d。联合使用FK5 0 6和ALS组的供体血细胞嵌合率停药后 3周稳定在 10 %以上。　结论　短期使用FK5 0 6和ALS可显著延长兔同种异体膝关节移植的存活时间 ,可维持 3周的嵌合耐受。     

去细胞处理对化学去细胞异体神经免疫原性的影响

目的研究化学去细胞异体神经的免疫学和制备方法,以期进一步降低同种异体神经移植的免疫反应。方法取SD大鼠的坐骨神经,经4．0％Triton X-100和3．0％脱氧胆酸钠消化进行去细胞处理。将化学去细胞和新鲜的SD大鼠坐骨神经植入Wistar大鼠皮下,通过免疫组化方法观察移植物及周围组织中的CD3、CIM和CD8阳性T细胞浸润程度,评价免疫反应强度。将犬的尺神经按上述方法进行去细胞处理,按去细胞处理次数分为3个组,分别从去细胞程度、髓鞘染色程度、GAG免疫组化染色和神经基底板层结构完整性方面进行组织学观察。结果与新鲜异体神经移植比较,化学去细胞异体神经植入后可明显降低大鼠的免疫原性,但仍有轻度细胞介导的免疫反应存在。经化学去细胞处理的神经,其残余神经轴突髓鞘的染色强度不随去细胞处理次数增加而明显减弱,GAG染色强度也无改变,但神经基底板层结构的破坏程度随去细胞处理次数的增加而增强。结论移植经化学去细胞处理的异体神经后,可有轻微的免疫反应,诱发免疫反应的物质可能与糖蛋白成分有关,残余的髓鞘组分不随去细胞处理次数增加而明显减少。     

心脏移植受者FK506治疗窗的临床初探

目的 寻求适合中国人心脏移植受者FK506理想治疗窗谷浓度范围。方法 应用微粒子酶免疫分析法（MEIA）测定6例心脏移植受者口服FK506后全血谷浓度,以FK506谷浓度结合病人临床疗效及不良反应的情况,总结FK506在心脏移植术后的治疗窗。结果 术后1年病人的FK506谷浓度控制在5-25ng/ml,未出现严重的排异反应和肾毒性,但术后早期曾出现头痛和震颤等不良反应。结论 FK506具有良好的免疫抑制效果,其治疗窗谷浓度范围,术后第1个月内为：15-20ng/ml,第2-3个月10-15ng/ml,第4-6个月8-12ng/ml,6个月后5-8ng/ml。此浓度范围即可有满意的免疫抑制效果。又可减少FK506的肾毒性。     

T细胞疫苗联合FK506抑制异种神经移植排斥的实验研究

目的 研究T细胞疫苗接种联合应用FK506对大鼠异种神经移植排斥反应的抑制作用. 方法 利用BABL/c小鼠的脾细胞免疫SD大鼠,取后者脾细胞活化所制备的T细胞疫苗对异种神经移植受体的SD大鼠进行免疫,移植后联合应用FK506.以异种神经移植后混合淋巴细胞培养、微量细胞毒测定、移植神经的组织学观察和胫前肌湿重测定等,对该方法抗大鼠异种神经移植排斥反应的作用加以分析. 结果 和对照组相比较,T细胞疫苗接种联合FK506使神经移植大鼠胫前肌湿重明显增加(P＜0.05),淋巴细胞转化率等排斥反应指标差异也具有统计学意义(P＜0.05). 结论 T细胞疫苗接种联合应用FK506能显著抑制大鼠异种神经移植的免疫排斥反应.     

FK506对周围神经端侧吻合侧枝生长的影响

目的 探讨FK506对神经端侧吻合侧枝生长的影响。方法 Wistar大白鼠26只，右侧腓总神经切断后，近端反转结扎，远端与胫神经外膜开窗处行端侧吻合，左侧作正常对照。术后动物分实验组和对照组，每组13只。于术后当天开始，实验组右小腿外侧肌肉注射生理盐水稀释的FK506（每日2mg／kg体重），连续2周，对照组注射等量的生理盐水。术后3个月，行双侧腓总神经、胫前肌组织学检查、胫前肌肌湿重测定及运动终板检查。结果 实验组及对照组腓总神经均有一定程度的再神经化，实验组再生神经纤维数目、肌湿重、肌纤维截面积、运动终板的面积及着色均明显优于对照组。结论FK506对神经端侧吻合侧枝生长具有促进作用。