猪肝星状细胞内质网应激模型的建立及对泛素化的影响

doi:10.16431/j.cnki.1671-7236.2020.02.010

Abstract

Abstract: This study was conducted to screen the different concentrations and culture time of tunicamycin (TM) in porcine immortalized hepatic stellate cell,in order to model endoplasmic reticulum stress (ERS) successfully and explore the changes of ubiquitination under ERS.Porcine hepatic stellate cell was treated with TM with concentrations of 0,1,2,5,10 and 15 μg/mL,respectively,and cultured for 2,4,8,16,24 and 36 h.TM concentration and culture time were initially screened by detection of cell inhibition rate,and finally selected by detection of cell cycle and ERS marker genes and proteins expression,in order to model ERS successfully.In addition,the expression of ubiquitin related genes in porcine liver stellate cells under ERS were detected.The results showed that 5 μg/mL TM culture for 8 or 24 h might model ERS via detection of cell inhibition rate.ERS marker gene expressions were all extremely significantly upregulated by 5 μg /mL TM,cultured for 8 or 24 h (P < 0.01).In terms of ERS marker protein expression,only ATF4 increased significantly (P < 0.05),and only G2/M phase of cell proportion in cell cycle significantly decreased at 8 h of culture (P < 0.05).At 24 h of culture,ERS marker proteins were significantly increased (P < 0.05),and cell cycle arrest occurred in G1 phase,leading to extremely significant declines in cell proportion in S phase and G2/M phase (P < 0.01).The result indicated that ERS model was successfully constructed treated by 5 μg/mL TM for 24 h.Ubiquitin related genes UBA2 and UBE2E expressions were extremely significantly increased (P < 0.01).In conclusion,the ERS model was successfully established and ubiquitylation mechanism was initiated by cultured with 5 μg/mL TM for 24 h in porcine hepatic stellate cell.

Key words: tunicamycin; porcine hepatic stellate cell; endoplasmic reticulum stress; ubiquitination

CLC Number:

Q256

XIN Hairui, ZHANG Chuang, HAO Yue, GU Xianhong. A Establishment of Endoplasmic Reticulum Stress Model of Porcine Hepatic Stellate Cell and Its Effect on Ubiquitination[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 399-406.

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A Establishment of Endoplasmic Reticulum Stress Model of Porcine Hepatic Stellate Cell and Its Effect on Ubiquitination

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