结缔组织生长因子抗体抑制慢性卡压后神经纤维化的研究
Inhibitory effects of connective tissue growth factor antibody on peripheral neural fibrosis after chronic entrapment
目的 观察结缔组织生长因子(CTGF)抗体对慢性卡压致神经纤维化的抑制作用.方法 将实验大鼠分为A、B、C3组.A组(假手术组):仅暴露坐骨神经;B组(单纯卡压组):依照Mackinnon建立坐骨神经慢性卡压模型设计的方法行坐骨神经套管卡压术;C组(卡压+抗体注射组):行坐骨神经卡压术,并在术中及术后3、6、9d定时注射CTGF多克隆抗体,A、B两组在相同部位注射等量的生理盐水做对照.于术后2、4、6、8、10周取神经行电镜、免疫组织化学、逆转录-聚合酶链反应(RT-PCR)、Western blot等检测方法分别检测不同时段卡压神经组织形态学变化及CTGF、Ⅰ型、Ⅲ型胶原蛋白(COL-Ⅰ、Ⅲ)含量.结果 坐骨神经单纯卡压组较假手术组CTGF、COL-Ⅰ、Ⅲ含量明显增多;CTGF多克隆抗体注射组神经形态学上纤维组织明显减少,COL-Ⅰ、Ⅲ含量降低,B组和C组神经中COL-Ⅰ mRNA表达量分别为:2周组(27.96±2.89、19.83±2.86)、4周组(32.29±3.58、24.18±3.17)、6周组(35.26±3.42、29.48±2.19),与单纯卡压组差异有统计学意义(P<0.05).结论 周围神经慢性卡压后可致神经纤维化,CTGF在此纤维化过程中起重要作用;CTGF抗体可有效抑制周围神经卡压后胶原合成,缓解慢性卡压后神经纤维化病变.
更多Objective To investigate the effect of connective tissue growth factor antibody on peripheral neural fibrosis after chronic entrapment.Methods Rats were randomly divided into three groups:group A (sham-operated),sciatic nerve exposed only; group B (compression only),subject to sciatic nerve entrapment operation according to the method described by Mackinnon; group C (compression + antibody injection),injection of CTGF polyclonal antibody 3,6 and 9 days respectively after sciatic nerve compression.The same volume of normal saline was given in groups A and B.Electron microscopy,immunohistochemistry,reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to observe the morphological changes of the compressed nerve tissue,and determine the level of CTGF,collagen-Ⅰ and collagen-Ⅲ at 2nd,4th,6th,8th,and 10th week after the surgery,respectively.Results The levels of CTGF,type-Ⅰ and Ⅲ collagen protein were increased significantly in group B as compared with group A.The expression of CTGF was increased in the early phase of the compression,and reached the peak at 4th week,and then decreased slowly.The expression of collagen-Ⅰ and collagen-Ⅲ was increased after compression,and reached the peak at 6th week,and then maintained a relatively-high level.Fibrous tissue was reduced significantly and the process of nerve regeneration accelerated as well as the levels of type-Ⅰ and Ⅲ collagen protein declined in group C,the expression levels of COL-Ⅰ mRNA in groups B and C were (27.96 ±2.89) and (19.83 ±2.86) at 2nd week,(32.29 ±3.58) and (24.18 ± 3.17) at 4th week,and (35.26 ±3.42) and (29.48 ± 2.19) at 6th week respectively,which were statistically significant from those in groups B and C (P < 0.05).Conclusion Peripheral nerve fibrosis can be caused by chronic nerve compression,and CTGF plays an important role in the process of fibrosis.CTGF antibody can effectively inhibit the synthesis of collagen protein following peripheral nervous entrapment so as to ameliorate nervous fibrosis.
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