Background and objectives This study focused on the effects of the combination of baicalin (BC) and gardenoside (GD) (7:3) on blood-brain barrier (BBB) permeability, brain tissue water content, and aquaporin-4 (AQP-4) expression in rats with cerebral ischemia-reperfusion (I/R) injury. The previous research conducted by the investigators demonstrated that the combination of BC and GD (7:3) has anti-ischemic properties. Further research was conducted to determine the mechanism underlying the reduction in cerebral edema.
Methods A total of 150 male Sprague-Dawley rats were randomly assigned to the following groups to receive treatment: sham, I/R, allyl chloride (AC), 30 mg/kg BC/GD, and 60 mg/kg BC/GD groups. Then, neurobehavioral scores were assigned to determine the effectiveness of the treatment. Evans blue (EB) was used to trace the BBB. The dry/wet method was used to evaluate the brain water content. Transmission electron microscopy was performed to examine the ultrastructure of the brain tissue. Immunohistochemistry and western blot were performed to detect the presence of AQP-4 in the hippocampus. Reverse transcription polymerase chain reaction (RT-PCR) was used to determine the amount of AQP-4 mRNA.
Results The BBB permeability, brain water content, and AQP-4 expression were significantly greater in the CA1 area of the hippocampus in the I/R group, when compared to the sham group. Furthermore, the endoplasmic reticulum was dilated, and most of the nerve cells underwent degeneration or necrosis. After the BC/GD treatment, the markers improved in a dose-dependent manner.
Conclusions BC/GD can inhibit the BBB permeability and cerebral edema by reducing the expression of AQP-4 in the CA1 area of the hippocampus in rats after I/R injury, improving the structure of nerve cells and exerting brain-protective effects.
Acknowledgments
We would like to thank International Science Editing for the lan-guage polishing of the manuscript.
Funding
This research was supported by grants from the National Natural Science Foundation of China (Grant no. 81473385), and the Subject Innovation Team of Shaanxi University of Chinese Medicine (Grant no. 2019-YL13).
Conflict of interest
The authors have no conflicts of interest related to this publication.
Author contributions
LZ: conceptualization, supervision, data curation, writing of the original draft, writing of the review, and editing. HHZ: investiga-tion and writing of the original draft. QQS: investigation and data curation. ADZ: investigation and data curation. CW: conceptual-ization. JPL: conceptualization. BW: conceptualization, supervi-sion, validation, writing of the review, and editing. All authors read and approved the final manuscript.
Ethical statement
The animal handling procedures and experimental protocols were consistent with the Guide for the Care and Use of Laboratory Ani-mals, and approved by the Animal Ethics Committee of Shaanxi University of Chinese Medicine. Ethics approval no.: SUCMDL 20220401004.
Data sharing statement
The data used to support the findings of the study are included in the article.
| [1] |
Moriyama Y, Takagi N, Itokawa C, Tanonaka K. Injection of neural progenitor cells attenuates decrease in level of connexin 43 in brain capillaries after cerebral ischemia. Neurosci Lett 2013; 543:152-156. doi:10.1016/j.neulet.2013.03.053,PMID:23583594.
|
| [2] |
Li YH, Qiao LJ, Lin XY. Mechanism of low molecular weight GTP bind-ing protein RAC1 in injury of neural function of rats with cerebral ischemia reperfusion. Asian Pac J Trop Med 2016; 9(5):474-477. doi: 10.1016/j.apjtm.2016.03.024,PMID:27261857.
|
| [3] |
Wang Y, He Z, Deng S. Ursolic acid reduces the metalloprotease/anti-metalloprotease imbalance in cerebral ischemia and reperfusion injury. Drug Des Devel Ther 2016; 10:1663-1674. doi:10.2147/DDDT.S103829,PMID:27274199.
|
| [4] |
Rosenberg GA. Ischemic brain edema. Prog Cardiovasc Dis 1999;42(3):209-216. doi:10.1016/s0033-0620(99)70003-4, PMID:105 98921.
|
| [5] |
Verkman AS, Smith AJ, Phuan PW, Tradtrantip L, Anderson MO. The aquaporin-4 water channel as a potential drug target in neurological disorders. Expert Opin Ther Targets 2017; 21(12):1161-1170. doi:10.1080/14728222.2017.1398236,PMID:29072508.
|
| [6] |
Wang C, Yan M, Jiang H, Wang Q, He S, Chen J, et al. Mechanism of aquaporin 4 (AQP 4) up-regulation in rat cerebral edema under hypobaric hypoxia and the preventative effect of puerarin. Life Sci 2018; 193:270-281. doi:10.1016/j.lfs.2017.10.021,PMID:29054452.
|
| [7] |
Sun K, Fan J, Han J. Ameliorating effects of traditional Chinese medi-cine preparation, Chinese materia medica and active compounds on ischemia/reperfusion-induced cerebral microcirculatory dis-turbances and neuron damage. Acta Pharm Sin B 2015; 5(1):8-24. doi:10.1016/j.apsb.2014.11.002,PMID:26579420.
|
| [8] |
Wang Y, Xiao G, He S, Liu X, Zhu L, Yang X, et al. Protection against acute cerebral ischemia/reperfusion injury by QiShenYiQi via neu-roinflammatory network mobilization. Biomed Pharmacother 2020; 125:109945. doi:10.1016/j.biopha.2020.109945,PMID:32028240.
|
| [9] |
Cao G, Jiang N, Hu Y, Zhang Y, Wang G, Yin M, et al. Ruscogenin Atten-uates Cerebral Ischemia-Induced Blood-Brain Barrier Dysfunction by Suppressing TXNIP/NLRP3 Inflammasome Activation and the MAPK Pathway. Int J Mol Sci 2016; 17(9):1418. doi:10.3390/ijms17091418,PMID:27589720.
|
| [10] |
Tang C, Xue H, Bai C, Fu R, Wu A. The effects of Tanshinone IIA on blood-brain barrier and brain edema after transient middle cerebral artery occlusion in rats. Phytomedicine 2010; 17(14):1145-1149. doi:10.1016/j.phymed.2010.03.017,PMID:20570121.
|
| [11] |
Wang P, Cao Y, Yu J, Liu R, Bai B, Qi H, et al. Baicalin alleviates is-chemia-induced memory impairment by inhibiting the phospho-rylation of CaMKII in hippocampus. Brain Res 2016; 1642:95-103. doi:10.1016/j.brainres.2016.03.019,PMID:27016057.
|
| [12] |
Yang G, Li JQ, Bo JP, Wang B, Tian XR, Liu TZ, et al. Baicalin inhibits PDGF-induced proliferation and migration of airway smooth muscle cells. Int J Clin Exp Med 2015; 8(11):20532-20539. PMID:26884970.
|
| [13] |
Zhang Z, Gao X, Guo M, Jiang H, Cao Y, Zhang N. The Protective Effect of Baicalin Against Lead-Induced Renal Oxidative Damage in Mice. Biol Trace Elem Res 2017; 175(1):129-135. doi:10.1007/s12011-016-0731-2,PMID:27209023.
|
| [14] |
Xue X, Qu XJ, Yang Y, Sheng XH, Cheng F, Jiang EN, et al. Baicalin attenuates focal cerebral ischemic reperfusion injury through in-hibition of nuclear factor κB p65 activation. Biochem Biophys Res Commun 2010; 403(3-4):398-404. doi:10.1016/j.bbrc.2010.11.042,PMID:21093411.
|
| [15] |
Zhou QB, Duan CZ, Jia Q, Liu P, Li LY. Baicalin attenuates focal cer-ebral ischemic reperfusion injury by inhibition of protease-activated receptor-1 and apoptosis. Chin J Integr Med 2014; 20(2):116-122. doi:10.1007/s11655-013-1441-7,PMID:23504578.
|
| [16] |
Liang H, Zhang L, Wang H, Tang J, Yang J, Wu C, et al. Inhibitory Ef-fect of Gardenoside on Free Fatty Acid-Induced Steatosis in HepG2 Hepatocytes. Int J Mol Sci 2015; 16(11):27749-27756. doi:10.3390/ijms161126058,PMID:26610473.
|
| [17] |
Liu AP, Li M, Bin W, Cao H, Li Y, Zhou Fy. Effect of baicalin and Jas-minoidin compatibility on cerebral ischemia-reperfusion injury Pharmacol Clin Chin Mater Med 2016; 32(01):46-48. doi:10.13412/j.cnki.zyyl.2016.01.013. (in Chinese).
|
| [18] |
Li M, Bin W, Tang Z, Hou J, Sun J. Effect of baicalin and geniposide on CysLT2R and CysLT2RmRNA expression in hippocampus of cerebral ischemia and reperfusion injury rats at acute and early recovery phase Pharmacol Clin Chin Mater Med 2012; 28(05):18-20. doi:10.13412/j.cnki.zyyl.2012.05.011. (in Chinese).
|
| [19] |
Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20(1):84-91. doi:10.1161/01.str.20.1.84,PMID:2643202.
|
| [20] |
Zhao X, Li S, Mo Y, Li R, Huang S, Zhang A, et al. DCA Protects against Oxidation Injury Attributed to Cerebral Ischemia-Reperfusion by Reg-ulating Glycolysis through PDK2-PDH-Nrf 2 Axis. Oxid Med Cell Lon-gevity 2021; 2021:5173035. doi:10.1155/2021/5173035,PMID:34712383.
|
| [21] |
Cao Y, Mao X, Sun C, Zheng P, Gao J, Wang X, et al. Baicalin attenuates global cerebral ischemia/reperfusion injury in gerbils via anti-oxida-tive and anti-apoptotic pathways. Brain Res Bull 2011; 85(6):396-402. doi:10.1016/j.brainresbull.2011.05.002, PMID:21600966.
|
| [22] |
Tan HM, Zhou L, Zou HC.Geniposide-mediated GLP-1R/Akt signaling pathway improves cerebral ischemia-reperfusion injury and neuronal apoptosis in rats Drug Eval Res 2022; 45(09):1822-1829. doi:10.7501/j.issn.1674-6376.2022.09.017. (in Chinese).
|
| [23] |
You LJ, Yang XF, Geng H, Meng JL, Ma YH, Lei M, et al. Inhibition of in-flammation of Geniposide on LPS induced RAW264.7 cells based on Caspase-1 cell pyroptosis signal pathway Chin J Immunol 2022; 38(13):1574-1578. doi:10.3969/j.issn.1000-484X.2022.13.007. (in Chinese).
|
| [24] |
Koo HJ, Lim KH, Jung HJ, Park EH. Anti-inflammatory evaluation of gardenia extract, geniposide and genipin. J Ethnopharmacol 2006; 103(3):496-500. doi:10.1016/j.jep.2005.08.011,PMID:16169698.
|
| [25] |
Wang J, Hou JC, Xiang LH, Zhang J, Ju DH. Compatibility of Geniposide and Ginsenoside Rg1: Their Regulating Roles in Secretion of Anoxia Induction Injured Microglia Inflammatory Cytokines. Chin J Integr Tra-dit West Med 2014; 34(01):91-95. doi:10.7661/CJIM.2014.01.0091.
|
| [26] |
Yao X, Derugin N, Manley GT, Verkman AS. Reduced brain edema and infarct volume in aquaporin-4 deficient mice after transient focal cerebral ischemia. Neurosci Lett 2015; 584:368-372. doi:10.1016/j.neulet.2014.10.040, PMID:25449874.
|
| [27] |
Xu SH, Yin MS, Liu B, Chen ML, He GW, Zhou PP, et al. Tetramethylpyrazine-2′-O-sodium ferulate attenuates blood-brain barrier disruption and brain oedema after cerebral is-chemia/reperfusion. Hum Exp Toxicol 2017; 36(7):670-680. doi:10.1177/0960327116657401,PMID:27387348.
|
| [28] |
Lu L, Li HQ, Li JH, Liu AJ, Zheng GQ. Neuroprotection of Sanhua De-coction against Focal Cerebral Ischemia/Reperfusion Injury in Rats through a Mechanism Targeting Aquaporin 4. Evidence-Based Comple-mentary Altern Med 2015; 2015:584245. doi:10.1155/2015/584245,PMID:26089944.
|
| [29] |
Mohammadi MT, Dehghani GA. Nitric oxide as a regulatory factor for aquaporin-1 and 4 gene expression following brain ischemia/reperfu-sion injury in rat. Pathol, Res Pract 2015; 211(1):43-49. doi:10.1016/j.prp.2014.07.014, PMID:25441658.
|
| [30] |
Vella J, Zammit C, Di Giovanni G, Muscat R, Valentino M. The cen-tral role of aquaporins in the pathophysiology of ischemic stroke. Front Cell Neurosci 2015;9:108. doi:10.3389/fncel.2015.00108,PMID:25904843.
|
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