TGF-β1-Mediated Activation of SERPINE1 is Involved in Hemin-Induced Apoptotic and Inflammatory Injury in HT22 Cells

Abstract

Tinggang Wang,1,* Haibin Lu,2,* Deqiang Li,2 Weichun Huang3 1Emergency Department, Affiliated Hospital of Zunyi Medical University, Zunyi, People’s Republic of China; 2Department of Critical Care Medicine, Daping Hospital, Army Medical University, Chongqing, People’s Republic of China; 3Radiology Department, First Affiliated Hospital of Army Medical University, Chongqing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weichun HuangRadiology Department, First Affiliated Hospital of Army Medical University, No. 30, Gao Tan Yan Main Street, Shapingba District, Chongqing, 400038, People’s Republic of ChinaTel [phone]Email [email protected]: Intracerebral hemorrhage (ICH) is a severe subtype of stroke with high mortality and morbidity. Serpin Family E Member 1 (SERPINE1) has been documented to be upregulated following ICH, however, the participation of SERPINE1 in the development of ICH has never been studied.Methods: Hemin was utilized to develop an in vitro model of ICH. Gene levels were evaluated by the use of quantitative reverse transcription polymerase chain reaction, Western blot, as well as enzyme-linked immunoassay assay. The activity of caspase-3 was determined using a commercial kit. Cell viability and apoptosis were assessed using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Terminal deoxynucleotidyl transferase (TdT) d UTP Nick-End Labeling assay.Results: SERPINE1 was upregulated in hemin-treated HT22 cells. Silencing of SERPINE1 attenuated hemin-induced inhibition of cell viability. Moreover, knockdown of SERPINE1 repressed hemin-induced apoptosis in HT22 cells, as evidenced by the decrease in the number of TUNEL positive cells, caspase-3 activity, and Bax expression, and the increase in Bcl-2 expression. Meanwhile, knockdown of SERPINE1 repressed hemin-induced inflammation in HT22 cells, as indicated by reduced levels of tumor necrosis factor-α, interleukin-6 (IL-6), IL-1β, and inducible nitric oxide synthase. We also found that transforming growth factor-beta 1 (TGF-β 1) induced SERPINE1 expression in a dose-dependent manner. Besides, SERPINE1 knockdown attenuated the effects of TGF-β 1 on hemin-induced neuronal damage.Conclusion: TGF-β 1-induced SERPINE1 activation exacerbated hemin-induced apoptosis and inflammation in HT22 cells, manifesting a novel mechanism for ICH progression.Keywords: intracerebral hemorrhage, serpin family E member 1, transforming growth factor-beta 1

Keywords

• intracerebral hemorrhage
• serpin family e member 1
• transforming growth factor-beta 1