Changes in expression of microRNA-221 and phosphatase and tension protein homologue in nerve stump after peripheral nerve injury_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Chenming ¹ , LI Yu ² , ZHANG Yuqiang ^1,3 , CAO Yu ⁴ , GONG Chao ¹ , WANG Chenliang ¹ ,  WANG Wei ^1,3,4

1. Department of Orthopedics, the First Affiliated Hospital of Jinzhou Medical University, Jinzhou Liaoning, 121000, P.R.China;
2. Graduate School of Liaoning University of Traditional Chinese Medicine, Shenyang Liaoning, 110016, P.R.China;
3. Liaoning Provincial Key Laboratory of Medical Tissue Engineering, Jinzhou Liaoning, 121000, P.R.China;
4. Institute of Extra-orbital Sciences, Jinzhou Medical University, Jinzhou Liaoning, 121000, P.R.China;

Corresponding?author：

WANG Wei, Email: weiwang_ly@126.com

Keywords：

Peripheral nerve injury; microRNA-221; phosphatase and tension protein homologue; nerve repair; rat

DOI：

10.7507/1002-1892.201903122

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Objective To study the expressions of microRNA-221 (miR-221) and the protein of phosphatase and tension protein homologue (PTEN) in the proximal and distal stumps after sciatic nerve injury in rats and their correlation with the repair of peripheral nerve injury, so as to provide a new target for clinical diagnosis of peripheral nerve injury.Methods Ninety-six male Sprague-Dawley rats of SPF grade were selected to establish sciatic nerve injury models. Twenty-four rats were sacrificed at 0 (immediately after operation), 1, 4, and 7 days after operation. The proximal and distal sciatic nerve fragments were taken under aseptic conditions. The expression of miR-221 was detected by real-time fluorescent quantitative PCR, and the expression of PTEN protein was detected by Western blot and immunofluorescent staining. The relationship between miR-221 and PTEN was verified by dual-luciferase reporter gene. At the same time, the ultrastructure of nerve stump was observed by transmission electron microscopy.Results The results of real-time fluorescent quantitative PCR, Western blot, and immunofluorescence staining showed that the relative expression of miR-221 in the proximal and distal stumps increased gradually with time, and the relative expression of PTEN protein decreased gradually, and the differences between different time points after operation were significant (P<0.05). At 1, 4, and 7 days after operation, the relative expression of miR-221 in proximal stump was significantly higher than that in distal stump, and the relative expression of PTEN protein in proximal stump was significantly lower than that in distal stump (P<0.05). Dual-luciferase reporter gene suggested that PTEN was the target for miR-221. Transmission electron microscopy observation showed that the normal morphological structure was observed at 0 day after operation, and the proliferation of Schwann cells and degeneration of axons and myelin sheaths gradually increased with time. There was no significant difference between proximal and distal stumps at 1 day after operation. At 4 and 7 days, Schwann cells proliferated more in proximal stump than in distal stump, and the degeneration of axons and myelin sheaths was less.Conclusion After sciatic nerve injury in rats, the up-regulation of the miR-221 expression targets the down-regulation of PTEN expression, which results in the difference of expression levels of miR-221 and PTEN in proximal and distal stumps. This phenomenon may play a role in promoting nerve repair after peripheral nerve injury.

Citation： ZHANG Chenming, LI Yu, ZHANG Yuqiang, CAO Yu, GONG Chao, WANG Chenliang, WANG Wei. Changes in expression of microRNA-221 and phosphatase and tension protein homologue in nerve stump after peripheral nerve injury. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(9): 1162-1168. doi: 10.7507/1002-1892.201903122 Copy

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2.	Sullivan R, Dailey T, Duncan K, et al. Peripheral nerve injury: stem cell therapy and peripheral nerve transfer. Int J Mol Sci, 2016, 17(12): E2101.
3.	Fernandez L, Komatsu DE, Gurevich M, et al. Emerging strategies on adjuvant therapies for nerve recovery. J Hand Surg (Am), 2018, 43(4): 368-373.
4.	林耀發, 宗海洋, 胡顯騰, 等. 大鼠坐骨神經損傷后 Spastin 表達變化的實驗研究. 中國修復重建外科雜志, 2017, 31(1): 86-90.
5.	Gallaher ZR, Steward O. Modest enhancement of sensory axon regeneration in the sciatic nerve with conditional co-deletion of PTEN and SOCS3 in the dorsal root ganglia of adult mice. Exp Neurol, 2018, 303: 120-133.
6.	Wong KM, Beirowski B. Multiple lines of inhibitory feedback on AKT kinase in Schwann cells lacking TSC1/2 hint at distinct functions of mTORC1 and AKT in nerve development. Commun Integr Biol, 2018, 11(1): e1433441.
7.	Krishnan A, Duraikannu A, Zochodne DW. Releasing " brakes” to nerve regeneration: intrinsic molecular targets. Eur J Neurosci, 2016, 43(3): 297-308.
8.	Shi J, Zhang Y, Jin N, et al. MicroRNA-221-3p plays an oncogenic role in gastric carcinoma by inhibiting PTEN expression. Oncol Res, 2017, 25(4): 523-536.
9.	Wang N, Zhu C, Xu Y, et al. Negative regulation of PTEN by microRNA-221 and its association with drug resistance and cellular senescence in lung cancer cells. Biomed Res Int, 2018, 2018: 7908950.
10.	Amini-Farsani Z, Sangtarash MH, Shamsara M, et al. MiR-221/222 promote chemoresistance to cisplatin in ovarian cancer cells by targeting PTEN/PI3K/AKT signaling pathway. Cytotechnology, 2018, 70(1): 203-213.
11.	Ye X, Bai W, Zhu H, et al. MiR-221 promotes trastuzumab-resistance and metastasis in HER2-positive breast cancers by targeting PTEN. BMB Rep, 2014, 47(5): 268-273.
12.	Bosse F. Extrinsic cellular and molecular mediators of peripheral axonal regeneration. Cell Tissue Res, 2012, 349(1): 5-14.
13.	Chen ZL, Yu WM, Strickland S. Peripheral regeneration. Annu Rev Neurosci, 2007, 30: 209-233.
14.	Jiang BG, Han N, Rao F, et al. Advance of peripheral nerve injury repair and reconstruction. Chin Med J (Engl), 2017, 130(24): 2996-2998.
15.	Savastano LE, Laurito SR, Fitt MR, et al. Sciatic nerve injury: a simple and subtle model for investigating many aspects of nervous system damage and recovery. J Neurosci Methods, 2014, 227: 166-180.
16.	馮思航, 馮登殿. 大鼠脛神經損傷后遠側端髓鞘和軸索再生的研究. 神經損傷與功能重建, 2016, 11(2): 103-106.
17.	Jessen KR, Mirsky R. Embryonic Schwann cell development: the biology of Schwann cell precursors and early Schwann cells. J Anat, 1997, 191(Pt 4): 501-505.
18.	李奕, 林巍巍, 王曉冬, 等. 成年大鼠周圍神經損傷后遠側端雪旺細胞變化的體外研究. 南通大學學報 (醫學版), 2006, 26(6): 414-416.
19.	魏壯, 李軍, 郭寶鋒, 等. 周圍神經損傷后近側端Schwann細胞變化的初步研究. 中國實驗診斷學, 2007, 11(2): 172-174.
20.	姚東東, 張潔元, 劉彬, 等. 周圍神經損傷修復微環境的研究進展. 中國修復重建外科雜志, 2015, 29(9): 1167-1172.
21.	Chen H, Xiang J, Wu J, et al. Expression patterns and role of PTEN in rat peripheral nerve development and injury. Neurosci Lett, 2018, 676: 78-84.
22.	Günzl P, Bauer K, Hainzl E, et al. Anti-inflammatory properties of the PI3K pathway are mediated by IL-10/DUSP regulation. J Leukoc Biol, 2010, 88(6): 1259-1269.
23.	Zhang J, Liu Y, Lu L. Emerging role of MicroRNAs in peripheral nerve system. Life Sci, 2018, 207: 227-233.
24.	Jiang N, Li H, Sun Y, et al. Differential gene expression in proximal and distal nerve segments of rats with sciatic nerve injury during Wallerian degeneration. Neural Regen Res, 2014, 9(12): 1186-1194.
25.	Qing L, Chen H, Tang J, et al. Exosomes and their microRNA cargo: new players in peripheral nerve regeneration. Neurorehabil Neural Repair, 2018, 32(9): 765-776.

1. Campbell WW. Evaluation and management of peripheral nerve injury. Clin Neurophysiol, 2008, 119(9): 1951-1965.
2. Sullivan R, Dailey T, Duncan K, et al. Peripheral nerve injury: stem cell therapy and peripheral nerve transfer. Int J Mol Sci, 2016, 17(12): E2101.
3. Fernandez L, Komatsu DE, Gurevich M, et al. Emerging strategies on adjuvant therapies for nerve recovery. J Hand Surg (Am), 2018, 43(4): 368-373.
4. 林耀發, 宗海洋, 胡顯騰, 等. 大鼠坐骨神經損傷后 Spastin 表達變化的實驗研究. 中國修復重建外科雜志, 2017, 31(1): 86-90.
5. Gallaher ZR, Steward O. Modest enhancement of sensory axon regeneration in the sciatic nerve with conditional co-deletion of PTEN and SOCS3 in the dorsal root ganglia of adult mice. Exp Neurol, 2018, 303: 120-133.
6. Wong KM, Beirowski B. Multiple lines of inhibitory feedback on AKT kinase in Schwann cells lacking TSC1/2 hint at distinct functions of mTORC1 and AKT in nerve development. Commun Integr Biol, 2018, 11(1): e1433441.
7. Krishnan A, Duraikannu A, Zochodne DW. Releasing " brakes” to nerve regeneration: intrinsic molecular targets. Eur J Neurosci, 2016, 43(3): 297-308.
8. Shi J, Zhang Y, Jin N, et al. MicroRNA-221-3p plays an oncogenic role in gastric carcinoma by inhibiting PTEN expression. Oncol Res, 2017, 25(4): 523-536.
9. Wang N, Zhu C, Xu Y, et al. Negative regulation of PTEN by microRNA-221 and its association with drug resistance and cellular senescence in lung cancer cells. Biomed Res Int, 2018, 2018: 7908950.
10. Amini-Farsani Z, Sangtarash MH, Shamsara M, et al. MiR-221/222 promote chemoresistance to cisplatin in ovarian cancer cells by targeting PTEN/PI3K/AKT signaling pathway. Cytotechnology, 2018, 70(1): 203-213.
11. Ye X, Bai W, Zhu H, et al. MiR-221 promotes trastuzumab-resistance and metastasis in HER2-positive breast cancers by targeting PTEN. BMB Rep, 2014, 47(5): 268-273.
12. Bosse F. Extrinsic cellular and molecular mediators of peripheral axonal regeneration. Cell Tissue Res, 2012, 349(1): 5-14.
13. Chen ZL, Yu WM, Strickland S. Peripheral regeneration. Annu Rev Neurosci, 2007, 30: 209-233.
14. Jiang BG, Han N, Rao F, et al. Advance of peripheral nerve injury repair and reconstruction. Chin Med J (Engl), 2017, 130(24): 2996-2998.
15. Savastano LE, Laurito SR, Fitt MR, et al. Sciatic nerve injury: a simple and subtle model for investigating many aspects of nervous system damage and recovery. J Neurosci Methods, 2014, 227: 166-180.
16. 馮思航, 馮登殿. 大鼠脛神經損傷后遠側端髓鞘和軸索再生的研究. 神經損傷與功能重建, 2016, 11(2): 103-106.
17. Jessen KR, Mirsky R. Embryonic Schwann cell development: the biology of Schwann cell precursors and early Schwann cells. J Anat, 1997, 191(Pt 4): 501-505.
18. 李奕, 林巍巍, 王曉冬, 等. 成年大鼠周圍神經損傷后遠側端雪旺細胞變化的體外研究. 南通大學學報 (醫學版), 2006, 26(6): 414-416.
19. 魏壯, 李軍, 郭寶鋒, 等. 周圍神經損傷后近側端Schwann細胞變化的初步研究. 中國實驗診斷學, 2007, 11(2): 172-174.
20. 姚東東, 張潔元, 劉彬, 等. 周圍神經損傷修復微環境的研究進展. 中國修復重建外科雜志, 2015, 29(9): 1167-1172.
21. Chen H, Xiang J, Wu J, et al. Expression patterns and role of PTEN in rat peripheral nerve development and injury. Neurosci Lett, 2018, 676: 78-84.
22. Günzl P, Bauer K, Hainzl E, et al. Anti-inflammatory properties of the PI3K pathway are mediated by IL-10/DUSP regulation. J Leukoc Biol, 2010, 88(6): 1259-1269.
23. Zhang J, Liu Y, Lu L. Emerging role of MicroRNAs in peripheral nerve system. Life Sci, 2018, 207: 227-233.
24. Jiang N, Li H, Sun Y, et al. Differential gene expression in proximal and distal nerve segments of rats with sciatic nerve injury during Wallerian degeneration. Neural Regen Res, 2014, 9(12): 1186-1194.
25. Qing L, Chen H, Tang J, et al. Exosomes and their microRNA cargo: new players in peripheral nerve regeneration. Neurorehabil Neural Repair, 2018, 32(9): 765-776.

Chinese Journal of Reparative and Reconstructive Surgery

Changes in expression of microRNA-221 and phosphatase and tension protein homologue in nerve stump after peripheral nerve injury

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