CTGF siRNA联合电针治疗抑制脊髓损伤胶质瘢痕形成

The study on CTGF siRNA combined with electro-acupuncture for inhibition of formation of glial scar following spinal cord injury

目的探讨抗结缔组织生长因子(CTGF)小干扰RNA(smal interferon RNA,siRNA)联合电针治疗对脊髓损伤(spinal cord injury,SCI)胶质瘢痕形成的影响。方法选用成年雌性SD大鼠,随机分为SCI组、电针(EA)组,CTGF siRNA干扰(CTGF)组、CTGF siRNA干扰联合电针(CTGF+EA)组,损伤后在3、7、14d分别取材,应用免疫荧光组织化学法、RTPCR技术,检测胶质纤维酸性蛋白(GFAP)、波形蛋白(Vimentin)、纤连蛋白(Fibronectin)、层连蛋白(Laminin)以及基因的表达变化。结果 SCI组星形胶质细胞反应性增生,数目增多;GFAP、Vimentin、Fibronectin、Laminin表达增加;EA组、CTGF组和CTGF+EA组GFAP、Vimentin、Fibronectin、Laminin表达降低,其中CTGF+EA组GFAP、Vimentin、Fibronectin、Laminin表达降低更显著。RT-PCR结果显示:与SCI组相比,EA组、CTGF组和CTGF+EA组GFAP、Vimentin、Fibronectin、Laminin基因mRNA表达均降低(P<0.05),与EA组、CTGF组相比,CTGF+EA组GFAP、Vimentin、Fibronectin、Laminin表达降低更明显(P<0.05)。结论脊髓损伤后,CTGF siRNA联合电针治疗能有效抑制GFAP、Vimentin、Fibronectin、Laminin的表达,在一定程度上能抑制胶质瘢痕的形成,有助于神经功能的恢复。

Objective To investigate the anti-CTGF small interferon RNA（siRNA）combined with electro-acupuncture for inhibition of formation of glial scar following spinal cord injury（SCI）.Methods Adult female SD rats were randomly divided into SCI group,electro-acupuncture（EA）group,CTGF siRNA interference（CTGF）group,and CTGF siRNA interference combined with electro-acupuncture（CTGF ＋ EA）group.They were sampled at 3d,7dand 14 dafter injury,respectively.The changes of glial scar following spinal cord injury were observed by using immunofluorescence histochemistry and reverse transcriptase polymerase chain reaction（RT-PCR）.The changes in the expression of GFAP,Vimentin,fibronectin（FN）and laminin（LN）,and mRNA expression of GFAP,Vimentin,FN and LN were detected,respectively.Results The SCI group showed reactive astrocyte proliferation with increased number;the expression levels of GFAP,Vimentin,FN and LN were increased.The expression levels of GFAP,Vimentin,FN and LN were decreased in the EA group、CTGF group and CTGF＋EA group..In CTGF ＋ EA group,the expression levels of GFAP,Vimentin,FN and LN were significantly decreased.RT-PCR results showed that the mRNA expression of GFAP,Vimentin,FN and LN in EA group、CTGF group and CTGF＋ EA group were all decreased compared with the SCI group（P0.05）;the expression of GFAP,Vimentin,FN and LN in the CTGF ＋ EA group were also decreased compared with the EA group、CTGF group（P0.05）.Conclusion After spinal cord injury,CTGF siRNA combined with electro-acupuncture can effectively inhibit the expression of GFAP,Vimentin,FN and LN,and can inhibit the formation of glial scar to a certain extent,contributing to the recovery of neurological function.