硫化氢调控BDNF-TrkB通路诱导HepG2细胞阿霉素耐受性

Hydrogen sulfide induces tolerance of HepG2 cells to adriamycin by modulation of BDNF-TrkB pathway

目的探讨硫化氢(hydrogen sulfide,H2S)对HepG2细胞阿霉素(adriamycin,ADM)耐受性产生的促进及其机制。方法以NaHS为H2S的供体,体外培养HepG2细胞,CCK-8法检测ADM对HepG2细胞的IC50值,碘化丙啶(propidium iodide,PI)染色后流式细胞仪检测细胞凋亡,Western blot检测HepG2细胞脑源性神经营养因子(brain derived neurotrophic factor,BDNF)的表达。结果 NaHS(100、200、400μmol·L-1)预处理HepG2细胞30 min能呈浓度依赖性地升高ADM处理72 h后对HepG2细胞的IC50值;200μmol·L-1NaHS预处理HepG2细胞30 min能明显降低0.32 mg·L-1ADM处理24 h后对HepG2细胞凋亡的诱导作用;NaHS(100、200、400μmol·L-1)作用HepG2细胞24h后,细胞BDNF的表达量明显增加;10 nmol·L-1BDNF受体(酪氨酸激酶受体B,tropomyosin-relatedkinase B,TrkB)阻断剂(K252a)预处理30 min可明显降低200μmol·L-1NaHS对HepG2细胞ADM IC50的增强作用和对ADM(0.32mg·L-1,24 h)诱导细胞凋亡的抑制作用。结论 H2S可促进HepG2细胞ADM耐受性的产生,其机制可能与其上调BDNF-TrkB通路有关。

Aim To investigate whether hydrogen sul- fide （H2S） induces the production of tolerance of HepG2 cells to adriamycin and the underlying mecha- nisms. Methods NariS was used as the donor of H2S. HepG2 cells were cultured in vitro. Cell Count- ing Kit-8 （CCK-8） assay was used to detect IC50 of ADM to HepG2 cells. The apoptosis of cells was detec- ted by flow cytometry （FCM） after propidium iodide （PI） staining. Western blot assay was used to detect the protein expression of brain derived neurotrophic factor （BDNF）. Results Pretreatment of HepG2 cells with NariS （ 100,200,400 μmol·L^-1 ） for 30 rain could notably increase IC50 of ADM to HepG2 cells in a concentration-dependent manner. Pretreatment of HepG2 cells with 200 μmol·L^-1 of H2S for 30 min significantly inhibited the apoptosis of HepG2 cells in-duced by treatment with 0.52 mg· L^-1 of ADM for 24 h. After treatment with NariS （ 100,200,400 μmol·L^-1 ） for 24 h, the expression of BDNF in HepG2 cells was markedly upregulated. Preteatment of HepG2 cells with 10 nmol · L^-1 of K252a, a specific inhibitor of tropomyosin-related kinase B （TrkB, the receptor of BDNF） for 30 min significantly reduced the increase in IC50 of adriamycin to HepG2 cells and the inhibition of ADM （0. 32 mg·L^-1, 24 h）-induced apoptosis of HepG2 cells caused by 200 μmol·L^-1 of NariS. Conclusion H2S can induce the production of toler- ance of HepG2 cells to adriamycin, which is involved in the upregulation of BDNF-TrkB pathway.