title: Chikusetsu Saponin IVa Ameliorates Cerebral Ischemia Reperfusion Injury in Diabetic Mice via Adiponectin-Mediated AMPK/GSK-3β Pathway In Vivo and In Vitro.
abstract: Diabetes mellitus substantially increases the risk of stroke and enhances brain's vulnerability to ischemia insult. In a previous study, Chikusetsu saponin IVa (CHS) pretreatment was proved to protect the brain from cerebral ischemic in normal stroke models. Whether CHS could attenuate cerebral ischemia/reperfusion (I/R) injury in diabetic mice and the possible underlying mechanism are still unrevealed. Male C57BL/6 mice were injected streptozotocin to induce diabetes. After that, the mice were pretreated with CHS for 1 month, and then, focal cerebral ischemia was induced following 24-h reperfusion. The neurobehavioral scores, infarction volumes, and some cytokines in the brain were measured. Apoptosis was analyzed by caspase-3, Bax, and Bcl-2 expression. Downstream molecules of adiponectin (APN) were investigated by Western blotting. The results showed that CHS reduced infarct size, improved neurological outcomes, and inhibited cell injury after I/R. In addition, CHS pretreatment increased APN level and enhanced neuronal AdipoR1, adenosine monophosphate-activated protein kinase (AMPK), and glycogen synthase kinase 3 beta (GSK-3β) expression in a concentration-dependent manner in diabetic mice, and these effects were abolished by APN knockout (KO). In vitro test, CHS treatment also alleviated PC12 cell injury and apoptosis, evidenced by reduced tumor necrosis factor alpha (TNF-α), malondialdehyde (MDA) and caspase-3 expression, and Bax/Bcl-2 ratio in I/R injured cells. Moreover, CHS enhanced AdipoR1, AMPK, and GSK-3β expression in a concentration-dependent manner. Likewise, short interfering RNA (sinRNA) knockdown of liver kinase B1 (LKB1), an upstream kinase of AMPK, reduced the ability of CHS in protecting cells from I/R injury. Furthermore, this LKB1-dependent cellular protection resulted from AdipoR1 and APN activation, as supported by the experiment using sinRNA knockdown of AdipoR1 and APN. Thus, CHS protected brain I/R in diabetes through AMPK-mediated phosphorylation of GSK-3β downstream of APN-LKB1 pathway.
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Jialin
authors: J
authors: Duan
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Ying
authors: Y
authors: Yin
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Jia
authors: J
authors: Cui
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Jiajia
authors: J
authors: Yan
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Yanrong
authors: Y
authors: Zhu
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Yue
authors: Y
authors: Guan
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Guo
authors: G
authors: Wei
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Yan
authors: Y
authors: Weng
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Xiaoxiao
authors: X
authors: Wu
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Chao
authors: C
authors: Guo
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China.
authors: Yanhua
authors: Y
authors: Wang
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China. handsomfish@aliyun.com.cn.
authors: Miaomiao
authors: M
authors: Xi
authors: Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, Changle West Road 127, Xi'an, Shaanxi, China. adwen-2012@hotmail.com.
authors: Aidong
authors: A
authors: Wen
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D052242:Adiponectin / Q000819:agonists / Q000378:metabolism*; D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism* / Q000517:prevention & control; D003921:Diabetes Mellitus, Experimental / Q000188:drug therapy / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D015427:Reperfusion Injury / Q000378:metabolism* / Q000517:prevention & control; D012503:Saponins / Q000494:pharmacology / Q000627:therapeutic use*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D052242:Adiponectin; D012503:Saponins; C098580:chikusetsu saponin IVa; D009828:Oleanolic Acid; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3; D055372:AMP-Activated Protein Kinases
keywords: AMPK; AdipoRs; Adiponectin; Chikusetsu saponin IVa; Diabetes; GSK-3β
references: Metab Brain Dis. 2015 Apr;30(2):437-47
references: 24737446
references: Biochem Biophys Res Commun. 1999 Apr 2;257(1):79-83
references: 10092513
references: Stroke. 2007 Mar;38(3):1044-9
references: 17272778
references: J Cereb Blood Flow Metab. 1998 Apr;18(4):367-75
references: 9538901
references: Biochem Pharmacol. 2006 May 28;71(11):1637-47
references: 16620785
references: Endocr Rev. 2005 May;26(3):439-51
references: 15897298
references: Hepatology. 2006 Feb;43(2 Suppl 1):S31-44
references: 16447272
references: Circulation. 2008 Jan 15;117(2):216-23
references: 18158361
references: Indian J Med Res. 2007 Mar;125(3):389-98
references: 17496363
references: J Cereb Blood Flow Metab. 1985 Jun;5(2):253-8
references: 3988824
references: Life Sci. 2013 Feb 7;92(2):131-6
references: 23201553
references: Cardiovasc Psychiatry Neurol. 2011;2011:609202
references: 21350721
references: N Engl J Med. 2002 Apr 11;346(15):1145-51
references: 11948275
references: Stroke. 2001 Oct;32(10):2426-32
references: 11588337
references: J Ethnopharmacol. 2014 Mar 28;152(3):553-60
references: 24524879
references: Shock. 2008 Sep;30(3):299-307
references: 18323734
references: Exp Brain Res. 1994;101(2):265-71
references: 7843312
references: Stroke. 2009 Jun;40(6):2157-64
references: 19372445
references: Clin J Am Soc Nephrol. 2007 Jul;2(4):631-6
references: 17699475
references: Life Sci. 2006 Aug 8;79(11):1040-8
references: 16674981
references: J Cell Physiol. 2012 Jan;227(1):239-49
references: 21412771
references: JAMA. 2013 Sep 4;310(9):948-59
references: 24002281
references: Neurology. 2002 Jul 9;59(1):67-71
references: 12105309
references: Brain Res. 2005 Aug 16;1053(1-2):1-9
references: 16038884
references: Circ Res. 2008 Feb 1;102(2):140-2
references: 18239140
references: Curr Diab Rep. 2007 Feb;7(1):25-33
references: 17254515
references: J Neurol Sci. 1976 Oct;29(2-4):267-75
references: 978211
references: Proc Natl Acad Sci U S A. 2004 Mar 9;101(10):3329-35
references: 14985505
references: Exp Transl Stroke Med. 2010 Aug 11;2(1):15
references: 20701790
references: J Endocrinol. 2009 Jan;200(1):93-105
references: 18971219
references: Hum Mol Genet. 2012 Jun 15;21(12 ):2725-37
references: 22419736
references: J Biol Chem. 2008 Dec 5;283(49):33902-10
references: 18801732
references: Mol Endocrinol. 2010 Jan;24(1):218-28
references: 19889816
references: Circulation. 2007 Mar 20;115(11):1408-16
references: 17339545
references: Nature. 2003 Jun 12;423(6941):762-9
references: 12802337
references: Neurology. 2005 Feb 22;64(4):654-9
references: 15728287
references: J Neurochem. 2009 May;109 Suppl 1:17-23
references: 19393004
references: J Clin Neurosci. 2002 Nov;9(6):618-26
references: 12604269
references: J Biol Chem. 2010 Oct 15;285(42):32182-91
references: 20688914
references: Lancet Neurol. 2012 Mar;11(3):261-71
references: 22341034
references: Ann Neurol. 1985 Mar;17(3):267-72
references: 3922283
references: Free Radic Res. 2003 May;37(5):471-80
references: 12797466
references: Biol Pharm Bull. 2007 Sep;30(9):1674-9
references: 17827719
references: PLoS One. 2013 Jun 07;8(6):e66382
references: 23762489
references: Eur J Pharmacol. 2009 Feb 14;604(1-3):111-6
references: 19135050
references: J Clin Endocrinol Metab. 2007 Mar;92(3):1129-36
references: 17213280
references: Diabetes Metab. 2010 Oct;36 Suppl 3:S31-8
references: 21211733
references: Diabetes. 2007 Jan;56(1):127-36
references: 17192474
references: Diabetes. 2002 Oct;51(10):2903-10
references: 12351425
references: Brain Res Brain Res Rev. 1997 Apr;23(3):210-8
references: 9164671
references: Eur J Neurosci. 2012 Oct;36(7):2899-905
references: 22817531
references: FASEB J. 2005 May;19(7):786-8
references: 15716393
references: Mol Cancer. 2008 May 12;7:37
references: 18474106
references: PLoS One. 2011;6(9):e24648
references: 21935433
references: Obesity (Silver Spring). 2006 Feb;14 Suppl 1:9S-15S
references: 16642957
references: J Stroke Cerebrovasc Dis. 2012 Nov;21(8):890-7
references: 21757374