法舒地尔促进小鼠皮瓣术后神经再生的研究  

Investigation of Fasudil in its efficacy at improving flap innervation in mice

作　　者：徐艺丹 王燕亭 陈绍锋 荆幸 徐丹 方芳[2] 王海[3] 谢昀[3] 庄跃宏[2] Xu Yidan;Wang yanting;Chen shaoFeng;Jing Xing;Xu Dan;Fangfang;Wang Hai;Xie Yun;Zhuang Yuehong(Department of Public Health and Medical Technology,Xiamen Medical College,Xiamen 361023,China;Fujian Provincial Key Laboratory of Brain Aging and Neurodegenerative Diseases,Institute of Clinical Applied Anatomy,Fujian Medical University,Fuzhou 350122,China;Department of traumatic Orthopedics,The First Affiliated Hospital of Fujian Medical University,Fuzhou 350122,China)

机构地区：[1]厦门医学院公共卫生与医学技术系,福建厦门361023 [2]福建医科大学福建省高校脑老化与神经变性疾病重点实验室,临床应用解剖学研究所,福州350122 [3]福建医科大学附属第一医院创伤骨科,福州350122

出　　处：《中国临床解剖学杂志》2023年第4期447-452,458,共7页Chinese Journal of Clinical Anatomy

基　　金：福建省自然科学资金(2020J01625,2021J01241,2021J01666);福建省教育厅教师教育科研项目(JAT200723);天然化妆品福建省高校工程研究中心(厦门医学院)开放课题(XMMC-NC202104)。

摘　　要：目的探讨法舒地尔是否能促进皮瓣术后神经再生及其机制。方法选用92只Institute of Cancer Research(ICR)小鼠,随机分为对照组、Fasudil组、LY294002组、Fasudil+LY294002组,术前3 d开始分别每日腹腔注射相应药物。在术后0、5、7、15和30 d,免疫荧光染色观察皮瓣内的轴突生长情况。术后5 d,取皮瓣组织,Western blot检测皮瓣内RhoA、ROCK1+2、p-CPI-17、p-MYPT、p-PTEN、p-PI3K、p-Akt的表达情况。其次,取15 d孕鼠,取出Dorsal root ganglion(DRGs),分为6组:对照组、Fasudil组、Fasudil+LY294002组、Chondroitin sulfate proteoglycan(CSPG)组、CSPG+Fasudil组及CSPG+Fasudil+LY294002组,培养5 d后测量DRGs轴突长度。结果术后0 d各组均可观察到丰富的神经纤维;术后7 d,各组皮瓣内神经纤维都完全退变;术后15 d和30 d,只有Fasudil组再次出现神经纤维。术后5 d,Fasudil组的p-CPI-17、p-MYPT的表达下调,p-PI3K、p-Akt的表达上调(P<0.001)。对照组、Fasudil组、Fasudil+LY294002组、CSPG组、CSPG+Fasudil组及CSPG+Fasudil+LY294002组,培养5 d后DRGs的轴突平均长度分别为(1.96±0.24)、(2.73±0.30)、(2.07±0.13)、(1.62±0.20)、(2.35±0.13)及(1.84±0.31)mm,差异有统计学意义(P<0.001)。结论法舒地尔通过抑制RhoA/ROCK通路,激活PI3K/Akt通路促进皮瓣神经术后神经再生。Objective To explore whether Fasudil can promote the regeneration of the sensory nerve after flap surgery and its underlying mechanism.Methods 92 ICR mice were selected and randomly divided into the control,Fasudil,LY294002,and Fasudil+LY294002 groups,and the corresponding drugs were injected intraperitoneally daily from 3d before surgery.At 0 d,5 d,7 d,15 d and 30 d after operation,immunofluorescence staining was used to observe the axon outgrowth in the flap.The flap tissue at 5d after surgery was collected,and Western blotting was used to detect the expressions of RhoA,ROCK1+2,p-CPI-17,p-MYPT,p-PTEN,p-PI3K,and p-Akt in the flap.Secondly,rats 15d in pregnancy were adopted,the DRGs on both sides of the spinal cord were harvested and divided into 6 groups:the control,Fasudil,Fasudil+LY294002,CSPG,CSPG+Fasudil and CSPG+Fasudil+LY294002 groups.After 5d of culture,the axon lengths of the DRGss in the 6 groups were measured.Results Abundant nerve fibers could be observed in each group at 0d after operation;at 7d after operation,all nerve fibers underwent complete degeneration;at 15d and 30d after operation,only the Fasudil group had nerve fibers again.p-CPI-17 and p-MYPT in Fasudil group were down-regulated,and p-PI3K and p-Akt were up-regulated 5 days after operation(P<0.001).The average axon lengths of DRGs in the control,Fasudil,Fasudil+LY294002,CSPG,CSPG+Fasudil,and CSPG+Fasudil+LY294002 groups after 5d of culture were(1.96±0.24)mm,(2.73±0.30)mm,(2.07±0.13)mm,(1.62±0.20)mm,(2.35±0.13)mm,and(1.84±0.31)mm,respectively.The differences among the groups were statistically significant(P<0.001).Conclusions Fasudil promotes nerve regeneration in flap after surgery by inhibiting the RhoA/ROCK pathway and activating the PI3K/Akt pathway.

关 键 词：皮瓣 神经再生 法舒地尔 RhoA/ROCK PI3K/AKT 

分 类 号：R322.85[医药卫生—人体解剖和组织胚胎学]