ROCK inhibitor Y‑27632 protects rats against cerebral ischemia/reperfusion‑induced behavioral deficits and hippocampal damage

  • Authors:
    • Lihe Li
    • Baoyang Liu
  • View Affiliations

  • Published online on: August 9, 2019     https://doi.org/10.3892/mmr.2019.10584
  • Pages: 3395-3405
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Cerebral ischemic injury is a major cause of death and long‑term disability worldwide that leads to neurological and behavioral deficits, and for which successful treatments are still lacking. Ras homolog family member A (RhoA) and Rho‑associated coiled‑coil containing protein kinase (ROCK) are associated with the growth of neurons and the movement of neuronal growth cones. RhoA/ROCK inhibitors have been demonstrated to promote the recovery of motor function following nerve injury, but the underlying mechanism requires further investigation. The present study aimed to investigate the effects of the ROCK inhibitor Y‑27632 on middle cerebral artery occlusion (MCAO)‑induced cerebral ischemic injury. Rats were randomly assigned to the Control, Y‑27632, MCAO + Vehicle or MCAO + Y‑27632 group. Firstly, infarct volume, cognitive ability and cerebral injury were assessed. Secondly, indicators of cerebral inflammation, oxidative stress and apoptosis were evaluated. Finally, the expression of recombinant glial fibrillary acidic protein (GFAP) and allograft inflammatory factor 1 (AIF1) in the brain were measured to assess the activation of astrocytes and microglia, respectively. The results showed that Y‑27632 effectively increased the survival rate and behavioral performance of rats, and attenuated the cerebral injury, oxidative stress and cerebral inflammation levels following MCAO. The disturbance in hippocampal neurons caused by MCAO was also alleviated following treatment with Y‑27632. Neuronal apoptosis was also decreased following Y‑27632 treatment, as demonstrated by the TUNEL assay and the expression levels of Caspases‑3, 8 and 9 and Bax/Bcl‑2 ratio. The levels of GFAP and AIF1 were increased by MCAO and further promoted by Y‑27632, indicating the activation of astrocytes and microglia. In conclusion, the present study offered evidence of a protective effect of Y‑27632 administration on cerebral ischemia/reperfusion induced behavioral and hippocampal damage by activating astrocytes and microglia.

References

1 

Iadecola C and Anrather J: Stroke research at a crossroad: Asking the brain for directions. Nat Neurosci. 14:1363–1368. 2011. View Article : Google Scholar : PubMed/NCBI

2 

Breuer L, Knott M, Struffert T, Kloska S, Kurka N, Schwab S, Dörfler A, Köhrmann M and Engelhorn T: Limited versus whole-brain perfusion for the indication of thrombolysis in the extended time window of acute cerebral ischemia. J Stroke Cerebrovasc Dis. 24:2491–2496. 2015. View Article : Google Scholar : PubMed/NCBI

3 

Xiong XY, Liu L and Yang QW: Refocusing neuroprotection in cerebral reperfusion era: New challenges and strategies. Front Neurol. 9:2492018. View Article : Google Scholar : PubMed/NCBI

4 

Panieri E, Gogvadze V, Norberg E, Venkatesh R, Orrenius S and Zhivotovsky B: Reactive oxygen species generated in different compartments induce cell death, survival, or senescence. Free Radic Biol Med. 57:176–187. 2013. View Article : Google Scholar : PubMed/NCBI

5 

Xue J, Zhang X, Zhang C, Kang N, Liu X, Yu J, Zhang N, Wang H, Zhang L, Chen R, et al: Protective effect of Naoxintong against cerebral ischemia reperfusion injury in mice. J Ethnopharmacol. 182:181–189. 2016. View Article : Google Scholar : PubMed/NCBI

6 

Julian L and Olson MF: Rho-associated coiled-coil containing kinases (ROCK): Structure, regulation, and functions. Small GTPases. 5:e298462014. View Article : Google Scholar : PubMed/NCBI

7 

Schmandke A, Schmandke A and Strittmatter SM: ROCK and Rho: Biochemistry and neuronal functions of Rho-associated protein kinases. Neuroscientist. 13:454–469. 2007. View Article : Google Scholar : PubMed/NCBI

8 

Gong H and Yang CY: Morphological and hydrodynamic correlations with increasing outflow facility by rho-kinase inhibitor Y-27632. J Ocul Pharmacol Ther. 30:143–153. 2014. View Article : Google Scholar : PubMed/NCBI

9 

Oda T, Kume T, Izumi Y, Ishihara K, Sugmimoto H and Akaike A: Na+/Ca2+ exchanger inhibitors inhibit neurite outgrowth in PC12 cells. J Pharmacol Sci. 116:128–131. 2011. View Article : Google Scholar : PubMed/NCBI

10 

Wang J, Li H, Yao Y, Ren Y, Lin J, Hu J, Zheng M, Song X, Zhao T, Chen YY, et al: β-elemene enhances GAP-43 expression and neurite outgrowth by inhibiting RhoA kinase activation in rats with spinal cord injury. Neuroscience. 383:12–21. 2018. View Article : Google Scholar : PubMed/NCBI

11 

Pang Y, Chai CR, Gao K, Jia XH, Kong JG, Chen XQ, Vatcher G, Chen JG and Yu AC: Ischemia preconditioning protects astrocytes from ischemic injury through 14-3-3γ. J Neurosci Res. 93:1507–1518. 2015. View Article : Google Scholar : PubMed/NCBI

12 

Zhao JJ, Hu JX, Lu DX, Ji CX, Qi Y, Liu XY, Sun FY, Huang F, Xu P and Chen XH: Soluble cpg15 from astrocytes ameliorates neurite outgrowth recovery of hippocampal neurons after mouse cerebral ischemia. J Neurosci. 37:1628–1647. 2017. View Article : Google Scholar : PubMed/NCBI

13 

Yang C, Zhang X, Fan H and Liu Y: Curcumin upregulates transcription factor Nrf2, HO-1 expression and protects rat brains against focal ischemia. Brain Res. 28:133–141. 2009. View Article : Google Scholar

14 

Jia D, Deng Y, Gao J, Liu X, Chu J and Shu Y: Neuroprotective effect of Panax notoginseng plysaccharides against focal cerebral ischemia reperfusion injury in rats. Int J Biol Macromol. 63:177–180. 2014. View Article : Google Scholar : PubMed/NCBI

15 

Ahmadi M, Rajaei Z, Hadjzadeh MA, Nemati H and Hosseini M: Crocin improves spatial learning and memory deficits in the Morris water maze via attenuating cortical oxidative damage in diabetic rats. Neurosci Lett. 642:1–6. 2017. View Article : Google Scholar : PubMed/NCBI

16 

Mueller BK, Mack H and Teusch N: Rho kinase, a promising drug target for neurological disorders. Nat Rev Drug Discov. 4:387–398. 2005. View Article : Google Scholar : PubMed/NCBI

17 

Tanaka T, Nishimura D, Wu RC, Amano M, Iso T, Kedes L, Nishida H, Kaibuchi K and Hamamori Y: Nuclear Rho kinase, ROCK2, targets p300 acetyltransferase. J Biol Chem. 281:15320–15329. 2006. View Article : Google Scholar : PubMed/NCBI

18 

Wen JY, Gao SS, Chen FL, Chen S, Wang M and Chen ZW: Role of CSE-produced H2S on cerebrovascular relaxation via RhoA-ROCK inhibition and cerebral ischemia-reperfusion injury in Mice. ACS Chem Neurosci. 10:1565–1574. 2019. View Article : Google Scholar : PubMed/NCBI

19 

Sun XQ, Chen S, Wang LF and Chen ZW: Total flavones of Rhododendron simsii Planch flower protect isolated rat heart from ischaemia-reperfusion injury and its mechanism of UTR-RhoA-ROCK pathway inhibition. J Pharm Pharmacol. 70:1713–1722. 2018. View Article : Google Scholar : PubMed/NCBI

20 

Chen F, Liu Z, Peng W, Gao Z, Ouyang H, Yan T, Ding S, Cai Z, Zhao B, Mao L and Cao Z: Activation of EphA4 induced by EphrinA1 exacerbates disruption of the blood-brain barrier following cerebral ischemia-reperfusion via the Rho/ROCK signaling pathway. Exp Ther Med. 16:2651–2658. 2018.PubMed/NCBI

21 

Yada T, Shimokawa H, Hiramatsu O, Kajita T, Shigeto F, Tanaka E, Shinozaki Y, Mori H, Kiyooka T, Katsura M, et al: Beneficial effect of hydroxyfasudil, a specific Rho-kinase inhibitor, on ischemia/reperfusion injury in canine coronary microcirculation in vivo. J Am Coll Cardiol. 45:599–607. 2005. View Article : Google Scholar : PubMed/NCBI

22 

Hamid SA, Bower HS and Baxter GF: Rho kinase activation plays a major role as a mediator of irreversible injury in reperfused myocardium. Am J Physiol Heart Circ Physiol. 292:H2598–H2606. 2007. View Article : Google Scholar : PubMed/NCBI

23 

Shin HK, Salomone S, Potts EM, Lee SW, Millican E, Noma K, Huang PL, Boas DA, Liao JK, Moskowitz MA and Ayata C: Rho-kinase inhibition acutely augments blood flow in focal cerebral ischemia via endothelial mechanisms. J Cereb Blood Flow Metab. 27:998–1009. 2007. View Article : Google Scholar : PubMed/NCBI

24 

Tiftik RN, Baskurt OK, Kul S and Buyukafsar K: The functional significance of the rho/rho-kinase pathway in human erythrocytes. Turk J Haematol. 31:168–174. 2014. View Article : Google Scholar : PubMed/NCBI

25 

Iwasaki H, Okamoto R, Kato S, Konishi K, Mizutani H, Yamada N, Isaka N, Nakano T and Ito M: High glucose induces plasminogen activator inhibitor-1 expression through Rho/Rho-kinase-mediated NF-kappaB activation in bovine aortic endothelial cells. Atherosclerosis. 196:22–28. 2008. View Article : Google Scholar : PubMed/NCBI

26 

Shen L, Black ED, Witkowski ED, Lencer WI, Guerriero V, Schneeberger EE and Turner JR: Myosin light chain phosphorylation regulates barrier function by remodeling tight junction structure. J Cell Sci. 119:2095–2106. 2006. View Article : Google Scholar : PubMed/NCBI

27 

Satoh S, Utsunomiya T, Tsurui K, Kobayashi T, Ikegaki I, Sasaki Y and Asano T: Pharmacological profile of hydroxy fasudil as a selective rho kinase inhibitor on ischemic brain damage. Life Sci. 69:1441–1453. 2001. View Article : Google Scholar : PubMed/NCBI

28 

Rikitake Y, Kim HH, Huang Z, Seto M, Yano K, Asano T, Moskowitz MA and Liao JK: Inhibition of Rho kinase (ROCK) leads to increased cerebral blood flow and stroke protection. Stroke. 36:2251–2257. 2005. View Article : Google Scholar : PubMed/NCBI

29 

Lu Z, Cheng D, Yin J, Wu R, Zhang G, Zhao Q, Wang N, Wang F and Liang M: Antithrombin III protects against contrast-induced nephropathy. EBioMedicine. 17:101–107. 2017. View Article : Google Scholar : PubMed/NCBI

30 

Yin J, Wang F, Kong Y, Wu R, Zhang G, Wang N, Wang L, Lu Z and Liang M: Antithrombin III prevents progression of chronic kidney disease following experimental ischaemic-reperfusion injury. J Cell Mol Med. 21:3506–3514. 2017. View Article : Google Scholar : PubMed/NCBI

31 

Chandra S, Romero MJ, Shatanawi A, Alkilany AM, Caldwell RB and Caldwell RW: Oxidative species increase arginase activity in endothelial cells through the RhoA/Rho kinase pathway. Br J Pharmacol. 165:506–519. 2012. View Article : Google Scholar : PubMed/NCBI

32 

Kahles T, Luedike P, Endres M, Galla HJ, Steinmetz H, Busse R, Neumann-Haefelin T and Brandes RP: NADPH oxidase plays a central role in blood-brain barrier damage in experimental stroke. Stroke. 38:3000–3006. 2007. View Article : Google Scholar : PubMed/NCBI

33 

Gao Z, Zhu Q, Zhang Y, Zhao Y, Cai L, Shields CB and Cai J: Reciprocal modulation between microglia and astrocyte in reactive gliosis following the CNS injury. Mol Neurobiol. 48:690–701. 2013. View Article : Google Scholar : PubMed/NCBI

34 

Cragnolini AB, Montenegro G, Friedman WJ and Masco DH: Brain-region specific responses of astrocytes to an in vitro injury and neurotrophins. Mol Cell Neurosci. 88:240–248. 2018. View Article : Google Scholar : PubMed/NCBI

35 

Williams A, Piaton G and Lubetzki C: Astrocytes-friends or foes in multiple sclerosis? Glia. 55:1300–1312. 2007. View Article : Google Scholar : PubMed/NCBI

36 

Guo D, Murdoch CE, Xu H, Shi H, Duan DD, Ahmed A and Gu Y: Vascular endothelial growth factor signaling requires glycine to promote angiogenesis. Sci Rep. 7:147492017. View Article : Google Scholar : PubMed/NCBI

37 

Okada S, Nakamura M, Katoh H, Miyao T, Shimazaki T, Ishii K, Yamane J, Yoshimura A, Iwamoto Y, Toyama Y and Okano H: Conditional ablation of Stat3 or Socs3 discloses a dual role for reactive astrocytes after spinal cord injury. Nat Med. 12:829–834. 2006. View Article : Google Scholar : PubMed/NCBI

38 

Dos Santos AA, Lopez-Granero C, Farina M, Rocha JBT, Bowman AB and Aschner M: Oxidative stress, caspase-3 activation and cleavage of ROCK-1 play an essential role in MeHg-induced cell death in primary astroglial cells. Food Chem Toxicol. 113:328–336. 2018. View Article : Google Scholar : PubMed/NCBI

39 

Fu PC, Tang RH, Yu ZY, Xie MJ, Wang W and Luo X: The Rho-associated kinase inhibitors Y27632 and fasudil promote microglial migration in the spinal cord via the ERK signaling pathway. Neural Regen Res. 13:677–683. 2018. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

October 2019
Volume 20 Issue 4

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Li, L., & Li, L. (2019). ROCK inhibitor Y‑27632 protects rats against cerebral ischemia/reperfusion‑induced behavioral deficits and hippocampal damage. Molecular Medicine Reports, 20, 3395-3405. https://doi.org/10.3892/mmr.2019.10584
MLA
Li, L., Liu, B."ROCK inhibitor Y‑27632 protects rats against cerebral ischemia/reperfusion‑induced behavioral deficits and hippocampal damage". Molecular Medicine Reports 20.4 (2019): 3395-3405.
Chicago
Li, L., Liu, B."ROCK inhibitor Y‑27632 protects rats against cerebral ischemia/reperfusion‑induced behavioral deficits and hippocampal damage". Molecular Medicine Reports 20, no. 4 (2019): 3395-3405. https://doi.org/10.3892/mmr.2019.10584