Zhigang Chang1, Yongqing Li, Wei He, Baoling Liu, Xiuzhen Duan, Ihab Halaweish, Ted Bambakidis, Baihong Pan, Yingjian Liang, Vahagn C Nikolian, Patrick Georgoff, Hasan B Alam. 1. From the Department of Surgical ICU (Z.C.), Beijing Hospital, Beijing, China; Department of Cardiothoracic Surgery (W.H.), Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China; Department of Surgery (Z.C., Y.L., W.H., B.L., I.H., T.B., B.P., Y.L., V.C.N., P.G., H.B.A.), University of Michigan Hospital, Ann Arbor, Michigan; The First Hospital (Y.L.), China Medical University, Shengyang, China; and Department of Pathology (X.D.), Loyola University Medical Center, Maywood, Illinois.
Abstract
BACKGROUND: We recently discovered that Tubastatin-A, a histone deacetylase (HDAC6) inhibitor, can improve survival in a rodent model of hemorrhagic shock (HS), but mechanisms remain poorly defined. In this study, we investigated whether Tubastatin-A could protect intestinal tight junction (TJ) in HS. METHODS: In an in-vivo study with Wistar-Kyoto rats, the rats underwent HS (40% blood loss) followed by Tubastatin-A (70 mg/kg) treatment, without fluid resuscitation. The experimental groups were (1) sham (no hemorrhage, no treatment), (2) control (hemorrhage, without treatment), and (3) treatment (hemorrhage with Tubastatin-A administration). Six hours after hemorrhage, ileum was harvested. Whole cell lysate were analyzed for acetylated α-tubulin (Ac-tubulin), total tubulin, acetylated histone 3 at lysine 9 (Ac-H3K9), β-actin, claudin-3 and zonula occludens 1 (ZO-1) proteins by Western blot. Histological effects of Tubastatin-A on small bowel were examined. In an in-vitro study, human intestinal epithelial cells (Caco-2) were divided into three groups: (1) sham (normoxia), (2) control (anoxia, no treatment), and (3) treatment (anoxia, treatment with Tubastatin-A). After 12 hours in an anoxia chamber, the cells were examined for Ac-tubulin and Ac-H3K9, cellular viability, cytotoxicity, claudin-3 and ZO-1 protein expression, and transwell permeability study. RESULTS: Tubastatin-A treatment significantly attenuated HS-induced decreases of Ac-tubulin, Ac-H3K9, ZO-1 and claudin-3 proteins in small bowel in-vivo (p < 0.05). In cultured Caco-2 cells, anoxia significantly decreased cellular viability (p < 0.001) and increased cytotoxicity (p < 0.001) compared to the sham group, while Tubastatin-A treatment offered significant protection (p < 0.0001). Moreover, expression of claudin-3 was markedly decreased in vitro compared to the sham group, whereas this was significantly attenuated by Tubastatin-A (p < 0.05). Finally, anoxia markedly increased the permeability of Caco-2 monolayer cells (p < 0.05), while Tubastatin-A significantly attenuated the alteration (p < 0.05). CONCLUSION: Inhibition of HDAC6 can induce Ac-tubulin and Ac-H3K9, promote cellular viability, and prevent the loss of intestinal tight junction proteins during HS and anoxia.
BACKGROUND: We recently discovered that Tubastatin-A, a histone deacetylase (HDAC6) inhibitor, can improve survival in a rodent model of hemorrhagic shock (HS), but mechanisms remain poorly defined. In this study, we investigated whether Tubastatin-A could protect intestinal tight junction (TJ) in HS. METHODS: In an in-vivo study with Wistar-Kyoto rats, the rats underwent HS (40% blood loss) followed by Tubastatin-A (70 mg/kg) treatment, without fluid resuscitation. The experimental groups were (1) sham (no hemorrhage, no treatment), (2) control (hemorrhage, without treatment), and (3) treatment (hemorrhage with Tubastatin-A administration). Six hours after hemorrhage, ileum was harvested. Whole cell lysate were analyzed for acetylated α-tubulin (Ac-tubulin), total tubulin, acetylated histone 3 at lysine 9 (Ac-H3K9), β-actin, claudin-3 and zonula occludens 1 (ZO-1) proteins by Western blot. Histological effects of Tubastatin-A on small bowel were examined. In an in-vitro study, human intestinal epithelial cells (Caco-2) were divided into three groups: (1) sham (normoxia), (2) control (anoxia, no treatment), and (3) treatment (anoxia, treatment with Tubastatin-A). After 12 hours in an anoxia chamber, the cells were examined for Ac-tubulin and Ac-H3K9, cellular viability, cytotoxicity, claudin-3 and ZO-1 protein expression, and transwell permeability study. RESULTS:Tubastatin-A treatment significantly attenuated HS-induced decreases of Ac-tubulin, Ac-H3K9, ZO-1 and claudin-3 proteins in small bowel in-vivo (p < 0.05). In cultured Caco-2 cells, anoxia significantly decreased cellular viability (p < 0.001) and increased cytotoxicity (p < 0.001) compared to the sham group, while Tubastatin-A treatment offered significant protection (p < 0.0001). Moreover, expression of claudin-3 was markedly decreased in vitro compared to the sham group, whereas this was significantly attenuated by Tubastatin-A (p < 0.05). Finally, anoxia markedly increased the permeability of Caco-2 monolayer cells (p < 0.05), while Tubastatin-A significantly attenuated the alteration (p < 0.05). CONCLUSION: Inhibition of HDAC6 can induce Ac-tubulin and Ac-H3K9, promote cellular viability, and prevent the loss of intestinal tight junction proteins during HS and anoxia.
Authors: Mark A Rivieccio; Camille Brochier; Dianna E Willis; Breset A Walker; Melissa A D'Annibale; Kathryn McLaughlin; Ambreena Siddiq; Alan P Kozikowski; Samie R Jaffrey; Jeffery L Twiss; Rajiv R Ratan; Brett Langley Journal: Proc Natl Acad Sci U S A Date: 2009-11-02 Impact factor: 11.205
Authors: Edwin F de Zoeten; Liqing Wang; Kyle Butler; Ulf H Beier; Tatiana Akimova; Hong Sai; James E Bradner; Ralph Mazitschek; Alan P Kozikowski; Patrick Matthias; Wayne W Hancock Journal: Mol Cell Biol Date: 2011-03-28 Impact factor: 4.272
Authors: Ting Zhao; Yongqing Li; Roderick T Bronson; Baoling Liu; George C Velmahos; Hasan B Alam Journal: Surgery Date: 2014-06-16 Impact factor: 3.982
Authors: Aaron M Williams; Isabel S Dennahy; Umar F Bhatti; Ben E Biesterveld; Nathan J Graham; Yongqing Li; Hasan B Alam Journal: Shock Date: 2019-09 Impact factor: 3.454
Authors: Umar F Bhatti; Aaron M Williams; Ranganath G Kathawate; Panpan Chang; Jing Zhou; Ben E Biesterveld; Zhenyu Wu; Julia Dahl; Baoling Liu; Yongqing Li; Hasan B Alam Journal: Trauma Surg Acute Care Open Date: 2019-09-17