Sains Malaysiana 50(11)(2021): 3313-3320

http://doi.org/10.17576/jsm-2021-5011-15

MiR-122-5p Attenuates Endothelial-to-Mesenchymal Transition Induced by Oxygen and Glucose Deprivation/Reperfusion

(MiR-122-5p Mengatenuasi Peralihan Endotelium-kepada-Mesenkim Teraruh oleh Kekurangan Oksigen dan Glukosa/Reperfusi)

HAIYAN YU¹, JINQIU XU², PENGYIN ZHU³ & CHUNYAN ZHANG²*

¹Intensive Care Unit, Shanghai Quyang Hospital, Shanghai, China

²Department of Pharmacy, Affiliated Hospital of Nantong University, Nantong, China

³Department of Pharmacy, Affiliated Hai’an Peoples’ Hospital of Nantong University, Nantong, China

Received: 11 January 2021/Accepted: 11 March 2021

ABSTRACT

Endothelial-to-mesenchymal transition (EndoMT) is a common phenomenon in vascular diseases, while the role of endothelial dysfunction in central vascular disease remains to be further investigated. MiR-122 is an inflammation-associated non-coding RNA that participates in multiple human disease, but whether miR-122 plays as a critical role in EndoMT induced by ischaemic stroke is unknown. Although BAI2 is known as a brain-specific inhibitor protein of angiogenesis, few studies of BAI2 examined EndoMT. This study investigated the mechanism of EndoMT and the miR-122/BAI2 axis in oxygen-glucose deprivation/reperfusion (OGD/R)-mediated EndoMT. A transient middle cerebral artery occlusion (tMCAO) model and OGD/R treatment were used to mimic the ischaemia-reperfusion injury. The colocalization of CD31 and α-SMA was elevated in the peri-infarct area of tMCAO mice. The expression of miR-122 was decreased in the peri-infarct area of tMCAO mice. Downregulation of miR-122, Occludin, and ZO-1 was observed in human brain microvascular endothelial cells (HBMECs) after OGD/R treatment, while α-SMA expression was increased in HBMECs after OGD/R treatment. MiR-122 overexpression reduced the decrease of Occludin and ZO-1 expression and the increase of α-SMA expression induced by OGD/R. MiR-122 negatively regulated BAI2 expression, and OGD/R treatment enhanced BAI2 expression. Knockdown the expression of BAI2 suppressed the decrease of Occludin and ZO-1 expression and the increase of α-SMA expression induced by OGD/R. In conclusion, miR-122 overexpression attenuates OGD/R-mediated EndoMT by targeting BAI2.

Keywords: BAI2; EndoMT; miR-122; OGD/R

ABSTRAK

Peralihan endotelium-kepada-mesenkim (EndoMT) adalah fenomena biasa untuk penyakit vaskular, manakala peranan disfungsi endotelium pada penyakit vaskular pusat masih perlu dikaji lebih lanjut. MiR-122 ialah RNA bukan pengekodan yang berkaitan dengan keradangan yang menyumbang kepada pelbagai penyakit manusia, tetapi sama ada miR-122 memainkan peranan penting dalam EndoMT yang disebabkan oleh strok iskemia tidak diketahui. Walaupun BAI2 dikenali sebagai protein angiogenesis perencat khusus otak, beberapa kajian BAI2 mengkaji EndoMT. Penyelidikan ini mengkaji mekanisme EndoMT dan paksi miR-122/BAI2 dalam kekurangan oksigen-glukosa/reperfusi (OGD/R)-pengantara EndoMT. Model oklusi arteri serebrum tengah sementara (tMCAO) dan rawatan OGD/R digunakan untuk meniru kecederaan reperfusi iskemia. Kolokalisasi CD31 dan α-SMA dinaikkan di kawasan peri-infark tikus tMCAO. Pengekspresan miR-122 telah menurun di kawasan peri-infark tikus tMCAO. Pengawalaturan rendah miR-122, Occludin dan ZO-1 diperhatikan dalam sel endotelium mikrovaskular otak manusia (HBMEC) selepas rawatan OGD/R, manakala pengekspresan α-SMA meningkat dalam HBMEC selepas rawatan OGD/R. Pengekspresan berlebihan MiR-122 mengurangkan penurunan pengekspresan Occludin dan ZO-1 dan peningkatan pengekspresan α-SMA yang disebabkan oleh OGD/R. MiR-122 mengawal pengekspresan BAI2 secara negatif dan rawatan OGD/R meningkatkan pengekspresan BAI2. Pengurangan pengekspresan BAI2 menindas penurunan pengekspresan Occludin dan ZO-1 serta peningkatan pengekspresan α-SMA yang disebabkan oleh OGD/R. Kesimpulannya, pengekspresan berlebihan miR-122 melemahkan OGD/R-pengantara EndoMT dengan menyasarkan BAI2.

Kata kunci: BAI2; EndoMT; miR-122; OGD/R

REFERENCES

Bernstein, D.L., Zuluaga-Ramirez, V., Gajghate, S., Reichenbach, N.L., Polyak, B., Persidsky, Y. & Rom, S. 2019. miR-98 reduces endothelial dysfunction by protecting blood-brain barrier (BBB) and improves neurological outcomes in mouse ischemia/reperfusion stroke model. Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism. 271678X19882264.

Brait, V.H., Miro-Mur, F., Perez-de-Puig, I., Notario, L., Hurtado, B., Pedragosa, J., Gallizioli, M., Jimenez-Altayo, F., Arbaizar-Rovirosa, M., Otxoa-de-Amezaga, A., Monteagudo, J., Ferrer-Ferrer, M., de la Rosa, X., Bonfill-Teixidor, E., Salas-Perdomo, A., Hernandez-Vidal, A., Garcia-de-Frutos, P., Lauzurica, P. & Planas, A.M. 2019. CD69 plays a beneficial role in ischemic stroke by dampening endothelial activation. Circulation Research 124(2): 279-291.

Chen, A.Q., Fang, Z., Chen, X.L., Yang, S., Zhou, Y.F., Mao, L., Xia, Y.P., Jin, H.J., Li, Y.N., You, M.F., Wang, X.X., Lei, H., He, Q.W. & Hu, B. 2019. Microglia-derived TNF-alpha mediates endothelial necroptosis aggravating blood brain-barrier disruption after ischemic stroke. Cell Death & Disease 10(7): 487.

Hamzei Taj, S., Kho, W., Riou, A., Wiedermann, D. & Hoehn, M. 2016. MiRNA-124 induces neuroprotection and functional improvement after focal cerebral ischemia. Biomaterials 91: 151-165.

Jeong, B.C., Kim, M.Y., Lee, J.H., Kee, H.J., Kho, D.H., Han, K.E., Qian, Y.R., Kim, J.K. & Kim, K.K. 2006. Brain-specific angiogenesis inhibitor 2 regulates VEGF through GABP that acts as a transcriptional repressor. FEBS Letters 580(2): 669-676.

Kee, H.J., Koh, J.T., Kim, M.Y., Ahn, K.Y., Kim, J.K., Bae, C.S., Park, S.S. & Kim, K.K. 2002. Expression of brain-specific angiogenesis inhibitor 2 (BAI2) in normal and ischemic brain: Involvement of BAI2 in the ischemia-induced brain angiogenesis. Journal of Cerebral Blood Flow and Metabolism: Official Journal of the International Society of Cerebral Blood Flow and Metabolism 22(9): 1054-1067.

Li, D.B., Liu, J.L., Wang, W., Luo, X.M., Zhou, X., Li, J.P., Cao, X.L., Long, J.G., Chen, X.H. & Qin, C. 2018. Plasma exosomal miRNA-122-5p and miR-300-3p as potential markers for transient ischaemic attack in rats. Frontiers in Aging Neuroscience 10: 24.

Li, M., Tang, Y., Wu, L., Mo, F., Wang, X., Li, H., Qi, R., Zhang, H., Srivastava, A. & Ling, C. 2017. The hepatocyte-specific HNF4alpha/miR-122 pathway contributes to iron overload-mediated hepatic inflammation. Blood 130(8): 1041-1051.

Li, W.A., Efendizade, A. & Ding, Y. 2017. The role of microRNA in neuronal inflammation and survival in the post ischemic brain: A review. Neurological Research. pp. 1-9.

Li, X., Wei, Y. & Wang, Z. 2018. microRNA-21 and hypertension. Hypertension Research 41(9): 649-661.

Liu, C., Zhao, L., Han, S., Li, J. & Li, D. 2015. Identification and functional analysis of microRNAs in mice following focal cerebral ischemia injury. International Journal of Molecular Sciences 16(10): 24302-24318.

Liu, Y., Zhang, J., Han, R., Liu, H., Sun, D. & Liu, X. 2015. Downregulation of serum brain specific microRNA is associated with inflammation and infarct volume in acute ischemic stroke. Journal of Clinical Neuroscience: Official Journal of the Neurosurgical Society of Australasia 22(2): 291-295.

Lv, B., Cheng, X., Sharp, F.R., Ander, B.P. & Liu, D.Z. 2018. MicroRNA-122 mimic improves stroke outcomes and indirectly inhibits NOS₂ after middle cerebral artery occlusion in rats. Frontiers in Neuroscience 12: 767.

Miao, C., Xiong, Y., Zhang, G. & Chang, J. 2018. MicroRNAs in idiopathic pulmonary fibrosis, new research progress and their pathophysiological implication. Experimental Lung Research 44(3): 178-190.

Mohr, A.M. & Mott, J.L. 2015. Overview of microRNA biology. Seminars in Liver Disease. 35(1): 3-11.

Pastukh, N., Meerson, A., Kalish, D., Jabaly, H. & Blum, A. 2019. Serum miR-122 levels correlate with diabetic retinopathy. Clinical and Experimental Medicine 19(2): 255-260.

Sorensen, S.S., Nygaard, A.B., Carlsen, A.L., Heegaard, N.H.H., Bak, M. & Christensen, T. 2017. Elevation of brain-enriched miRNAs in cerebrospinal fluid of patients with acute ischemic stroke. Biomarker Research 5: 24.

Stanzione, R., Bianchi, F., Cotugno, M., Marchitti, S., Forte, M., Busceti, C., Ryskalin, L., Fornai, F., Volpe, M. & Rubattu, S. 2017. A decrease of brain microRNA-122 level is an early marker of cerebrovascular disease in the stroke-prone spontaneously hypertensive rat. Oxidative Medicine and Cellular Longevity 2017: 1206420.

Tan, S., Shan, Y., Lin, Y., Liao, S., Zhang, B., Zeng, Q., Wang, Y., Deng, Z., Chen, C., Hu, X., Peng, L., Qiu, W. & Lu, Z. 2019. Neutralization of interleukin-9 ameliorates experimental stroke by repairing the blood-brain barrier via down-regulation of astrocyte-derived vascular endothelial growth factor-A. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology 33(3): 4376-4387.

Vijayan, M., Alamri, F.F., Al Shoyaib, A., Karamyan, V.T. & Reddy, P.H. 2019. Novel miRNA PC-5P-12969 in ischemic stroke. Molecular Neurobiology 56(10): 6976-6985.

Wang, H.J., Wei, J.Y., Liu, D.X., Zhuang, S.F., Li, Y., Liu, H., Ban, M., Fang, W.G., Cao, L., Zhao, W.D. & Chen, Y.H. 2018. Endothelial Atg7 deficiency ameliorates acute cerebral injury induced by ischemia/reperfusion. Frontiers in Neurology 9: 998.

Warner, J.J., Harrington, R.A., Sacco, R.L. & Elkind, M.S.V. 2019. Guidelines for the early management of patients with acute ischemic stroke: 2019 Update to the 2018 guidelines for the early management of acute ischemic stroke. Stroke 50(12): 3331-3332. doi. 10.1161/STROKEAHA.119.027708.

Zhang, Y.P., Cui, Q.Y., Zhang, T.M., Yi, Y., Nie, J.J., Xie, G.H. & Wu, J.H. 2019. Chloroquine pretreatment attenuates ischemia-reperfusion injury in the brain of ob/ob diabetic mice as well as wildtype mice. Brain Research 146518.

*Corresponding author; email: ccyyzhangntfy@163.com