title: Secretory pathway Ca(2+)-ATPase isoform 1 knockdown promotes Golgi apparatus stress injury in a mouse model of focal cerebral ischemia-reperfusion: In vivo and in vitro study.
abstract: The present study was designed to investigate the potential role of secretory pathway Ca(2+)-ATPase isoform 1(SPCA1) in experimental focal cerebral ischemia-reperfusion injury. Cerebral ischemia-reperfusion was induced by transient middle cerebral artery occlusion (MCAO) for 2h s in Sprague-Dawley rats, and then the expression levels of SPAC1 mRNA and protein were determined. Results showed that SPCA1 level was transiently increased 1 day after reperfusion in peri-infarction area, while markedly increased in infarction core on 3day and 7 day after reperfusion. Then a SPCA1 lentivirus was used to achieve knockdown of SPCA1 gene: Ca(2+) transporting type 2C, member 1 (ATP2C1) gene. It has been observed that SPCA1 knockdown by lentivirus markedly increased cerebral infarction volume in vivo. Meanwhile, SPCA1 knockdown also facilitated per-oxidative production, including nitric oxide (NO) and 3-nitrotyrosine (3-NT) and decreased the expression of total superoxide dismutase (SOD) and manganese superoxide dismutase (MnSOD). Moreover, in vitro study showed that SPCA1 knockdown increased hydrogen peroxide (H2O2)-induced lactate dehydrogenase (LDH) leakage dose-dependently, and elevated caspase3 level in neuro-2a (N2a) cells. In addition, SPCA1 knockdown increased H2O2-induced production of nitric oxide and 3-NT dose-dependently, and reversed the increased activity of total SOD and MnSOD in neuro-2a cells. In conclusion, the present study indicated that SPCA1 could suppress over active Golgi apparatus (GA) stress thus attenuate cerebral ischemia-reperfusion injury.
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Changjie
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Wenna
authors.initials: W
authors.lastname: Peng
authors.affiliation: Department of Neurology, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yin
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Kong
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Chunna
authors.initials: C
authors.lastname: Lan
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Rehabilitation, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
authors.forename: Rumi
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Neurology, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China. Electronic address: huzhiping2014@163.com.
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Hu
keywords: Cerebral ischemia–reperfusion; Golgi apparatus stress; MCAO/R; Nitric oxide; OGD/R; SPCA1