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Yang-Jie Zhao,Huang Xingrui,Wen Fasheng,Huang Xinglong,Liu Zhixiao,Zhang Youxiang 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.2
Glutathione S-transferases (GSTs) are multifunctional detoxification enzymes which play important roles in protecting the organisms from environmental stress and are widely used as biomarkers for environmental bio monitoring studies. In this study, We investigated the GST activity in Protohermes costalis larvae, aquatic insects mainly found in oligotrophic fresh water environments, in response to cadmium (Cd) exposure. Eight GST genes in the larvae were identified and their expression patterns under Cd stress were determined by real-time quantitative polymerase chain reaction (RT-qPCR). The GST activity in the whole body was up-regulated by CdCl 2 in a dose-dependent manner and the midgut and malpighian tubules may be the main sites involved in GST activity regulation. The identified PcGSTs are members of cytosolic GST family containing the conserved glutathione-binding domain and substrate-binding domain. Six of them are highly enriched in malpighian tu bules, midgut or/and fat body. The expression level of PcGSTe1 was significantly up-regulated by 0.05, 0.1, 0.5, and 1 mM CdCl 2 treatments when compared with no Cd control. PcGSTs1 expression was significantly higher in 0.05 mM CdCl 2 and lower in 1 mM CdCl 2 compared with no Cd control. Other PcGSTs were up-regulated by different concentrations of CdCl 2 . Our results suggested that P. costalis midgut and malpighian tubules may be the main sites for GST activity regulation that induced by Cd in aqueous phase and increasing expression levels of different PcGST genes may be responsible for the GST activity up-regulation. Moreover, GST activity and gene expression in this insect may be used as biomarkers for future aquatic biomonitoring studies.
Ji Ningfei,Chen Zhongqi,Wang Zhengxia,Sun Wei,Yuan Qi,Zhang Xijie,Jia Xinyu,Wu Jingjing,Jiang Jingxian,Song Meijuan,Xu Tingting,Liu Yanan,Ma Qiyun,Sun Zhixiao,Bao Yanmin,Zhang Mingshun,Huang Mao 대한천식알레르기학회 2024 Allergy, Asthma & Immunology Research Vol.16 No.1
Purpose: The roles and mechanisms of long noncoding RNAs (lncRNAs) in T helper 2 (Th2) differentiation from allergic asthma are poorly understood. We aimed to explore a novel lncRNA, LincR-protein phosphatase 2 regulatory subunit B' gamma (PPP2R5C), in Th2 differentiation in a mouse model of asthma. Methods: LincR-PPP2R5C from RNA-seq data of CD4+ T cells of asthma-like mice were validated and confirmed by quantitative reverse transcription polymerase chain reaction, northern blotting, nuclear and cytoplasmic separation, and fluorescence in situ hybridization (FISH). Lentiviruses encoding LincR-PPP2R5C or shRNA were used to overexpress or silence LincR-PPP2R5C in CD4+ T cells. The interactions between LincR-PPP2R5C and PPP2R5C were explored with western blotting, chromatin isolation by RNA purification assay, and fluorescence resonance energy transfer. An ovalbumin-induced acute asthma model in knockout (KO) mice (LincR-PPP2R5C KO, CD4 conditional LincR-PPP2R5C KO) was established to explore the roles of LincR-PPP2R5C in Th2 differentiation. Results: LncR-PPP2R5C was significantly higher in CD4+ T cells from asthmatic mice ex vivo and Th2 cells in vitro. The lentivirus encoding LincR-PPP2R5C suppressed Th1 differentiation; in contrast, the short hairpin RNA (shRNA) lentivirus decreased LincR-PPP2R5C and Th2 differentiation. Mechanistically, LincR-PPP2R5C deficiency suppressed the phosphatase activity of the protein phosphatase 2A (PP2A) holocomplex, resulting in a decline in Th2 differentiation. The formation of an RNA-DNA triplex between LincR-PPP2R5C and the PPP2R5C promoter enhanced PPP2R5C expression and activated PP2A. LincR-PPP2R5C KO and CD4 conditional KO decreased Th2 differentiation, airway hyperresponsiveness and inflammatory responses. Conclusions: LincR-PPP2R5C regulated PPP2R5C expression and PP2A activity by forming an RNA-DNA triplex with the PPP2R5C promoter, leading to Th2 polarization in a mouse model of acute asthma. Our data presented the first definitive evidence of lncRNAs in the regulation of Th2 cells in asthma.