Effects of mitochondrial reactive oxygen species inhibitor R(+)-pramipexole on JAK2-STAT3 signaling pathway and pro-inflammatory factor TNF-α in rats with cerebral ischemia/reperfusion injury

doi:10.3969/j.issn.1006-7795.2021.02.012

Abstract

Abstract: Objective To study the effect of mitochondrial reactive oxygen species (ROS) inhibitor R(+)-pramipexole [R(+)-PPX] on the janus kinase 2 (JAK2)-signal transducer and activator of transcription 3 (STAT3) signaling pathway and pro-inflammatory factor tumor necrosis factor-α (TNF-α) in middle cerebral artery occlusion (MCAO) rats after 6 h reperfusion, and further to investigate the protective mechanism of R(+)-PPX on cerebral ischemia injury at early stage of reperfusion.Methods Thirty healthy male Sprague-Dawley rats were divided into Sham group, MCAO group, and R(+)-PPX group (n=10), according to random number table. MCAO rat model was established with modified suture method, the rats underwent 90 min of right MCAO and then reperfusion by withdrawing the filament. The anus temperature of the rats was monitored during the operation. The rats were sacrificed after 6h reperfusion, and then the brain was collected quickly. The level of p-JAK2 protein in ischemic brain tissue was determined with Western blotting. The expressions of p-STAT3 and TNF-α in the ischemic penumbra of frozen sections of brain tissues were detected with immunofluorescence staining. The ROS positive cells in the ischemic penumbra was detected with dihydroethidium (DHE) staining. Colocalization of ROS with p-STAT3 or TNF-α was examined with double labeling immunofluorescence staining in the ischemic penumbra. Colocalization of p-STAT3 and TNF-α was analyzed with double labeling immunofluorescence staining. Results 1) Compared with Sham group, the level of p-JAK2 protein was upregulated obviously in ischemic brain tissue of MCAO rats after 6 h reperfusion (P<0.05). The level of p-JAK2 protein decreased significantly in the ischemic brain tissue of R(+)-PPX group compared with that of MCAO group (P<0.05). 2) No p-STAT3 positive cell was observed in Sham group. The p-STAT3 positive cells in the ischemic penumbra of MCAO group increased obviously after 6 h reperfusion compared with Sham group (P<0.05). The p-STAT3 positive cells decreased significantly in the ischemic penumbra of R(+)-PPX group compared with MCAO group (P<0.05). ROS was colocalized with p-STAT3 in the ischemic penumbra of rats. 3) No TNF-α positive cell was observed in Sham group. The TNF-α positive cells in the ischemic penumbra of MCAO group increased obviously after 6 h reperfusion compared with Sham group (P<0.05). The TNF-α positive cells decreased significantly in the ischemic penumbra of R(+)-PPX group compared with MCAO group (P<0.05). ROS was colocalized with TNF-α in the ischemic penumbra of rats. 4) p-STAT3 was colocalized with TNF-α with immunofluorescence staining in the ischemic penumbra of MCAO group after 6 h reperfusion. Conclusion The mitochondrial ROS inhibitor R(+)-PPX may play a neuroprotective role at early stage of cerebral ischemia/reperfusion injury via inhibiting the activation of the JAK2-STAT3 signaling pathway and reducing the level of TNF-α in ischemic brain tissue of MCAO rats after 6 h reperfusion.

Key words: cerebral ischemia/reperfusion, mitochondrial reactive oxygen species, R(+)-PPX, janus kinase 2-signal transducer and activator of transcription 3 signaling pathway, tumor necrosis factor-α

CLC Number:

R743.3

Ding Mao, Yang Nan, Huang Yuyou, Shi Wenjuan, Yan Feng, Zhao Yongmei, Liu Kejian. Effects of mitochondrial reactive oxygen species inhibitor R(+)-pramipexole on JAK2-STAT3 signaling pathway and pro-inflammatory factor TNF-α in rats with cerebral ischemia/reperfusion injury[J]. Journal of Capital Medical University, 2021, 42(2): 232-238.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2021.02.012

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2021/V42/I2/232

References

[1] Zhao Y, Yan F, Yin J, et al.Synergistic interaction between zinc and reactive oxygen species amplifies ischemic brain injury in rats[J]. Stroke, 2018, 49(9): 2200-2210.
[2] Mo Y, Tang L, Ma Y, et al.Pramipexole pretreatment attenuates myocardial ischemia/reperfusion injury through upregulation of autophagy[J]. Biochem Biophys Res Commun, 2016, 473(4): 1119-1124.
[3] 尹洁,闫峰,罗玉敏,等. R(+)-普拉克索对大鼠短暂性局灶性脑缺血损伤的神经保护作用[J]. 首都医科大学学报,2014,35(2): 225-230.
[4] Tang Y, Tong X, Li Y,et al.JAK2/STAT3 pathway is involved in the protective effects of epidermal growth factor receptor activation against cerebral ischemia/reperfusion injury in rats[J]. Neurosci Lett, 2018, 662: 219-226.
[5] Charras A, Arvaniti P, Le Dantec C, et al.JAK inhibitors and oxidative stress control[J]. Front Immunol, 2019, 10: 2814.
[6] Krylatov A V, Maslov L N, Voronkov N S,et al.Reactive oxygen species as intracellular signaling molecules in the cardiovascular system[J]. Curr Cardiol Rev, 2018, 14(4): 290-300.
[7] Chae U, Kim H S, Lee H S, et al.Drp1-dependent mitochondrial fission regulates p62-mediated autophagy in LPS-induced activated microglial cells[J]. Biosci Biotechnol Biochem, 2019, 83(3): 409-416.
[8] Li T, Wu S, Zhang H, et al.Activation of nicotinic receptors inhibits TNF-alpha-Induced production of pro-inflammatory mediators through the JAK2/STAT3 signaling pathway in fibroblast-like synoviocytes[J]. Inflammation, 2015, 38(4): 1424-1433.
[9] Zhang Y P, Zhang Y, Xiao Z B, et al.CFTR prevents neuronal apoptosis following cerebral ischemia reperfusion via regulating mitochondrial oxidative stress[J]. J Mol Med (Berl), 2018, 96(7): 611-620.
[10] Hu G, Du X, Li Y, et al.Inhibition of cerebral ischemia/reperfusion injury-induced apoptosis: nicotiflorin and JAK2/STAT3 pathway[J]. Neural Regen Res, 2017, 12(1): 96-102.
[11] Lin H W, Liu C W, Yang D J, et al.Dunaliella salina alga extract inhibits the production of interleukin-6, nitric oxide, and reactive oxygen species by regulating nuclear factor-kappaB/Janus kinase/signal transducer and activator of transcription in virus-infected RAW264.7 cells[J]. J Food Drug Anal, 2017, 25(4): 908-918.
[12] Wu L, Tan J L, Wang Z H, et al.ROS generated during early reperfusion contribute to intermittent hypobaric hypoxia-afforded cardioprotection against postischemia-induced Ca(2+) overload and contractile dysfunction via the JAK2/STAT3 pathway[J]. J Mol Cell Cardiol, 2015, 81: 150-161.
[13] Evzelman M A, Mityaeva E V, Lashkhiia I B,et al.Acute cerebral ischemia and inflammation[J]. Zh Nevrol Psikhiatr Im S S Korsakova, 2019, 119(12. Vyp. 2): 73-80.
[14] Hu Z, Yu F, Gong P, et al.Subneurotoxic copper(II)-induced NF-κB-dependent microglial activation is associated with mitochondrial ROS[J]. Toxicol Appl Pharmacol, 2014, 276(2): 95-103.
[15] Ye J, Chen D, Ye Z, et al.Fucoidan isolated from saccharina japonica inhibits LPS-induced inflammation in macrophages via blocking NF-kappaB, MAPK and JAK-STAT pathways[J]. Mar Drugs, 2020, 18(6):328.