CAJ | 학술논문

目的：：通过体外实验探讨硫醚环化 B 螺旋肽(thioether-cyclized helix B peptide,CHBP)对肾小管上皮细胞的保护作用及机制。方法：体外培养人近端肾小管上皮细胞(human renal proximal tubular cell line,HK-2)并分为 Control 组、H2 O2组和 CHBP 组。H2 O2组和 CHBP 组均通过500μmol/L H2 O2刺激4 h 建立氧化应激损伤模型。CHBP 组在建立氧化应激损伤模型1 h 前加入20 nmol/L 的 CHBP。500μmol/L H2 O2刺激4 h 后,检测细胞的活性、凋亡情况以及氧化应激损伤程度。采用 Western blotting 和 PCR 技术检测连接免疫球蛋白(binding immunoglobulin protein,BiP)、C/EBP 同源蛋白(C/EBP ho-mologous protein,CHOP)以及下游 NFE2相关因子2(NF-E2-related factor 2,Nrf2)信号通路中的分子表达水平,判断内质网应激水平。结果：CHBP 预处理可以减轻氧化应激导致的细胞毒性损伤；CCK-8试剂盒和原位末端标记法(TUNEL)检测发现,CHBP 预处理后细胞活性增加,凋亡减少,氧化应激和内质网应激水平下降。CHBP 组 Nrf2和血红素氧合酶1(heme oxygenase-1,HO-1)水平较 H2 O2组显著升高。结论：CHBP 预处理可以减轻 HK-2细胞的氧化应激损伤,这一保护作用可能是通过抑制内质网应激和激活下游 Nrf2信号通路实现的。
목적：：통과체외실험탐토류미배화 B 라선태(thioether-cyclized helix B peptide,CHBP)대신소관상피세포적보호작용급궤제。방법：체외배양인근단신소관상피세포(human renal proximal tubular cell line,HK-2)병분위 Control 조、H2 O2조화 CHBP 조。H2 O2조화 CHBP 조균통과500μmol/L H2 O2자격4 h 건립양화응격손상모형。CHBP 조재건립양화응격손상모형1 h 전가입20 nmol/L 적 CHBP。500μmol/L H2 O2자격4 h 후,검측세포적활성、조망정황이급양화응격손상정도。채용 Western blotting 화 PCR 기술검측련접면역구단백(binding immunoglobulin protein,BiP)、C/EBP 동원단백(C/EBP ho-mologous protein,CHOP)이급하유 NFE2상관인자2(NF-E2-related factor 2,Nrf2)신호통로중적분자표체수평,판단내질망응격수평。결과：CHBP 예처리가이감경양화응격도치적세포독성손상；CCK-8시제합화원위말단표기법(TUNEL)검측발현,CHBP 예처리후세포활성증가,조망감소,양화응격화내질망응격수평하강。CHBP 조 Nrf2화혈홍소양합매1(heme oxygenase-1,HO-1)수평교 H2 O2조현저승고。결론：CHBP 예처리가이감경 HK-2세포적양화응격손상,저일보호작용가능시통과억제내질망응격화격활하유 Nrf2신호통로실현적。
Objective:To explore the protective effect of thioether-cyclized helix B peptide (CHBP)on renal tubular epithelial cells and its mechanism by experiment in vitro.Methods:Human renal proximal tubular epithelial cell line HK-2 cells were cul-tured in vitro and divided into control group,H2 O2 group and CHBP group.Oxidative stress damage models were established by stimulation with 500 μmol/L H2 O2 for 4 h.In CHBP group,20 nmol/L CHBP was added to the culture medium 1 h prior to the establishment of oxidative stress model.After 4 h stimulation with 500 μmol/L H2 O2 ,cell viability,situation of apopto-sis,and severity of oxidative stress damage were measured.The expressions of Binding immunoglobulin protein(BiP),C/EBP homologous protein(CHOP),and the molecular in downstream NF-E2-related factor 2 (Nrf2)signaling pathway were detected by Western blotting and real-time PCR analysis,so as to evaluate the level of endoplasmic reticulum(ER)stress.Results:Oxi-dative stress induced cytotoxic injury could be alleviated by CHBP pretreatment.Kit-8 (CCK-8 )assay and TUNEL showed that,after CHBP pretreatment,cell viability increased,while cell apoptosis,oxidative stress and ER stress decreased.The levels of Nrf2 and heme oxygenase-1(HO-1)were significantly higher in CHBP group than those in H2 O2 group.Conclusions:CHBP pretreatment could alleviate oxidative stress damage of HK-2 cells,and the protective effect may be achieved by inhibi-ting ER stress and activating downstream Nrf2 signaling pathway.