Xiao-Xu-Ming decoction extract ameliorates brain injury in rats with thrombotic focal ischemic stroke and understanding possible therapeutic targets using proteomics

doi:10.5246/jcps.2021.06.036

Abstract

Abstract:

Ischemic stroke seriously threatens human health and quality of life. Xiao-Xu-Ming (XXM) decoction has been a classical prescription for stroke therapy. In our previous studies, we have found that XXM exerts neuroprotective effects, improves brain injury, and attenuates neuroinflammation in cerebral ischemia rats. In this study, we investigated the effects and possible mechanism of XXM on thrombotic focal cerebral ischemia. After treatment with XXM, the neurological function and motor abilities were improved, and cerebral infarction volume was significantly decreased compared with rats of thrombotic focal cerebral ischemia. Besides, the results of BBB integrity detected by EB leakage and tight junction (TJ) protein expression showed that XXM could maintain BBB integrity and improve the expressions of TJ proteins, including claudin-1, occluding, and ZO-1, in the ischemic ipsilateral cortex disrupted by thrombotic cerebral ischemia. Furthermore, proteomic techniques were used to identify the differentially expressed proteins (DEPs) in the ischemic cerebral cortex, and the results showed that 132 DEPs regulated by XXM were detected in the ischemic cerebral cortex. Bioinformatic analysis showed that these regulated proteins by XXM were mainly involved in complement and coagulation cascade, and lysosome, etc. Furthermore, there was an interaction among DEPs, including Lgals3, Ctsz, Capg, C1qa, S100a4, Grn, Hspb1, Aif1, and Anxa1, etc. In conclusion, XXM ameliorated brain injury of thrombotic focal ischemic stroke, and Lgals3, Ctsz, Capg, C1qa, S100a4, Grn, Hspb1, Aif1, and Anxa1 could help provide possible therapeutic targets of XXM for ischemic stroke and offer research direction for further research.

Key words: Xiao-Xu-Ming decoction, Thrombotic focal ischemic stroke, Blood-brain barrier, Proteomics

Supporting:

Yinglin Yang, Shanshan Zhang, Man Liu, Yuehua Wang, Guanhua Du. Xiao-Xu-Ming decoction extract ameliorates brain injury in rats with thrombotic focal ischemic stroke and understanding possible therapeutic targets using proteomics[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(6): 468-483.

Figures/Tables 8

Table 1. Primer list for RT-PCR.

References 35

[1]	Bernheisel, C.R.; Schlaudecker, J.D.; Leopold, K. Subacute management of ischemic stroke. Am. Fam. Physician. 2011, 84, 1383–1388.
[2]	Barthels, D.; Das, H. Current advances in ischemic stroke research and therapies. Biochim Biophys. Acta Mol. Basis. Dis. 2020, 1866, 165260.
[3]	Cai, Z.; Qiao, P.F.; Wan, C.Q.; Cai, M.; Zhou, N.K.; Li, Q. Role of blood-brain barrier in Alzheimer’s disease. J. Alzheimers. Dis. 2018, 63, 1223–1234.
[4]	Keaney, J.; Campbell, M. The dynamic blood-brain barrier. FEBS J. 2015, 282, 4067–4079.
[5]	Li, W.H.; Cheng, X.; Yang, Y.L.; Liu, M.; Zhang, S.S.; Wang, Y.H.; Du, G.H. Kaempferol attenuates neuroinflammation and blood brain barrier dysfunction to improve neurological deficits in cerebral ischemia/reperfusion rats. Brain Res. 2019, 1722, 146361.
[6]	Tsai, T.H.; Song, E.; Zhu, R.; Di Poto, C.; Wang, M.; Luo, Y.; Varghese, R.S.; Tadesse, M.G.; Ziada, D.H.; Desai, C.S.; Shetty, K.; Mechref, Y.; Ressom, H.W. LC-MS/MS-based serum proteomics for identification of candidate biomarkers for hepatocellular carcinoma. Proteomics. 2015, 15, 2369–2381.
[7]	Wang, Y.H.; Yang, Y.L.; Cheng, X.; Zhang, J.; Li, W.; Du, G.H. Xiao-Xu-Ming decoction extract regulates differentially expressed proteins in the hippocampus after chronic cerebral hypoperfusion. Neural. Regen. Res. 2019, 14, 470–479.
[8]	Jia, Z.; Tie, C.; Wang, C.; Wu, C.; Zhang, J. Perturbed lipidomic profiles in rats with chronic cerebral ischemia are regulated by Xiao-xu-Ming decoction. Front. Pharmacol. 2019, 10, 264.
[9]	Cheng, X.; Yang, H.; Yang, Y.L.; Li, W.H.; Liu, M.; Wang, Y.H.; Du, G.H. Xiao-Xu-Ming decoction extract alleviates LPS-induced neuroinflammation associated with down-regulating TLR4/MyD88 signaling pathway in vitro and in vivo. J. Chin. Pharm. Sci. 2019, 28, 88–99.
[10]	Wang, Y.H.; He, X.L.; Yang, H.G.; Qin, H.L.; Du, G.H. Effects of the effective components group of Xiao-xu-ming Decoction on MCAO rats. Chin. Pharm. J. 2012, 47, 194–198.
[11]	Lan, R.; Xiang, J.; Wang, G.H.; Li, W.W.; Zhang, W.; Xu, L.L.; Cai, D.F. Xiao-xu-Ming decoction protects against blood-brain barrier disruption and neurological injury induced by cerebral ischemia and reperfusion in rats. Evid Based Complement. Alternat. Med. 2013, 2013, 629782.
[12]	Lan, R.; Zhang, Y.; Xiang, J.; Zhang, W.; Wang, G.H.; Li, W.W.; Xu, L.L.; Cai, D.F. Xiao-Xu-Ming decoction preserves mitochondrial integrity and reduces apoptosis after focal cerebral ischemia and reperfusion via the mitochondrial p53 pathway. J. Ethnopharmacol. 2014, 151, 307–316.
[13]	Zhang, R.; Liu, C.; Ji, Y.Q.; Teng, L.; Guo, Y.L. Neuregulin-1β plays a neuroprotective role by inhibiting the Cdk5 signaling pathway after cerebral ischemia-reperfusion injury in rats. J. Mol. Neurosci. 2018, 66, 261–272.
[14]	Liu, P.; Tang, Y.Y.; Yang, X.S.; Dai, J.; Yang, M.; Zhang, H.; Liu, Y.; Yan, H.; Song, X.Y. Validation of a preclinical animal model to assess brain recovery after acute stroke. Eur. J. Pharmacol. 2018, 835, 75–81.
[15]	Cheng, X.; Yang, Y.L.; Li, W.H.; Liu, M.; Wang, Y.H.; Du, G.H. Cerebral ischemia-reperfusion aggravated cerebral infarction injury and possible differential genes identified by RNA-Seq in rats. Brain Res. Bull. 2020, 156, 33–42.
[16]	Cheng, X.; Yang, Y.L.; Li, W.H.; Liu, M.; Zhang, S.S.; Wang, Y.H.; Du, G.H. Dynamic alterations of brain injury, functional recovery, and metabolites profile after cerebral ischemia/reperfusion in rats contributes to potential biomarkers. J. Mol. Neurosci. 2020, 70, 667–676.
[17]	Bárez-López, S.; Bosch-García, D.; Gómez-Andrés, D.; Pulido-Valdeolivas, I.; Montero-Pedrazuela, A.; Obregon, M.J.; Guadaño-Ferraz, A. Abnormal motor phenotype at adult stages in mice lacking type 2 deiodinase. PLoS One. 2014, 9, e103857.
[18]	Wu, G.; McBride, D.W.; Zhang, J.H. Axl activation attenuates neuroinflammation by inhibiting the TLR/TRAF/NF-κB pathway after MCAO in rats. Neurobiol. Dis. 2018, 110, 59–67.
[19]	Sommer, C.J. Ischemic stroke: experimental models and reality. Acta Neuropathol. 2017, 133, 245–261.
[20]	Li, W.H.; Yang, Y.L.; Cheng, X.; Liu, M.; Zhang, S.S.; Wang, Y.H.; Du, G.H. Baicalein attenuates caspase-independent cells death via inhibiting PARP-1 activation and AIF nuclear translocation in cerebral ischemia/reperfusion rats. Apoptosis. 2020, 25, 354–369.
[21]	Grysiewicz, R.A.; Thomas, K.; Pandey, D.K. Epidemiology of ischemic and hemorrhagic stroke: incidence, prevalence, mortality, and risk factors. Neurol. Clin. 2008, 26, 871–95, vii.
[22]	Campbell, B.C. Thrombolysis and thrombectomy for acute ischemic stroke: strengths and synergies. Semin. Thromb. Hemost. 2017, 43, 185–190.
[23]	McBride, D.W.; Zhang, J.H. Precision stroke animal models: the permanent MCAO model should be the primary model, not transient MCAO. Transl. Stroke Res. 2017, 8, 397–404.
[24]	Abdullahi, W.; Tripathi, D.; Ronaldson, P.T. Blood-brain barrier dysfunction in ischemic stroke: targeting tight junctions and transporters for vascular protection. Am. J. Physiol. Cell Physiol. 2018, 315, C343–C356.
[25]	Sladojevic, N.; Stamatovic, S.M.; Johnson, A.M.; Choi, J.; Hu, A.N.; Dithmer, S.; Blasig, I.E.; Keep, R.F.; Andjelkovic, A.V. Claudin-1-dependent destabilization of the blood-brain barrier in chronic stroke. J. Neurosci. 2019, 39, 743–757.
[26]	Jiao, H.X.; Wang, Z.H.; Liu, Y.H.; Wang, P.; Xue, Y.X. Specific role of tight junction proteins claudin-5, occludin, and ZO-1 of the blood-brain barrier in a focal cerebral ischemic insult. J. Mol. Neurosci. 2011, 44, 130–139.
[27]	Walther, T.C.; Mann, M. Mass spectrometry-based proteomics in cell biology. J. Cell Biol. 2010, 190, 491–500.
[28]	Satyam, A.; Graef, E.R.; Lapchak, P.H.; Tsokos, M.G.; Dalle Lucca, J.J.; Tsokos, G.C. Complement and coagulation cascades in trauma. Acute. Med. Surg. 2019, 6, 329–335.
[29]	Comte, I.; Kim, Y.; Young, C.C.; van der Harg, J.M.; Hockberger, P.; Bolam, P.J.; Poirier, F.; Szele, F.G. Galectin-3 maintains cell motility from the subventricular zone to the olfactory bulb. J. Cell Sci. 2011, 124, 2438–2447.
[30]	James, R.E.; Hillis, J.; Adorján, I.; Gration, B.; Mundim, M.V.; Iqbal, A.J.; Majumdar, M.M.; Yates, R.L.; Richards, M.M.; Goings, G.E.; DeLuca, G.C.; Greaves, D.R.; Miller, S.D.; Szele, F.G. Loss of galectin-3 decreases the number of immune cells in the subventricular zone and restores proliferation in a viral model of multiple sclerosis. Glia. 2016, 64, 105–121.
[31]	Lalancette-Hébert, M.; Swarup, V.; Beaulieu, J.M.; Bohacek, I.; Abdelhamid, E.; Weng, Y.C.; Sato, S.; Kriz, J. Galectin-3 is required for resident microglia activation and proliferation in response to ischemic injury. J. Neurosci. 2012, 32, 10383–10395.
[32]	Sirko, S.; Irmler, M.; Gascón, S.; Bek, S.; Schneider, S.; Dimou, L.; Obermann, J.; De Souza Paiva, D.; Poirier, F.; Beckers, J.; Hauck, S.M.; Barde, Y.A.; Götz, M. Astrocyte reactivity after brain injury-: The role of galectins 1 and 3. Glia. 2015, 63, 2340–2361.
[33]	Young, C.C.; Al-Dalahmah, O.; Lewis, N.J.; Brooks, K.J.; Jenkins, M.M.; Poirier, F.; Buchan, A.M.; Szele, F.G. Blocked angiogenesis in Galectin-3 null mice does not alter cellular and behavioral recovery after middle cerebral artery occlusion stroke. Neurobiol. Dis. 2014, 63, 155–164.
[34]	Lerman, B.J.; Hoffman, E.P.; Sutherland, M.L.; Bouri, K.; Hsu, D.K.; Liu, F.T.; Rothstein, J.D.; Knoblach, S.M. Deletion of galectin-3 exacerbates microglial activation and accelerates disease progression and demise in a SOD1(G93A) mouse model of amyotrophic lateral sclerosis. Brain Behav. 2012, 2, 563–575.
[35]	Jiang, H.R.; Al Rasebi, Z.; Mensah-Brown, E.; Shahin, A.; Xu, D.M.; Goodyear, C.S.; Fukada, S.Y.; Liu, F.T.; Liew, F.Y.; Lukic, M.L. Galectin-3 deficiency reduces the severity of experimental autoimmune encephalomyelitis. J. Immunol. 2009, 182, 1167–1173.