【摘要】 目的探讨尿酸对原代培养的乳鼠心肌成纤维细胞(CFs)胶原合成及炎性分泌的影响,以及microRNA-155(miR-155)在其中的作用。方法分离培养乳鼠CFs,并用不同浓度(0、200、400、600、800μmol/L)的尿酸处理。采用MTT法检测CFs的增殖情况,羟脯氨酸试剂盒检测胶原含量,酶联免疫吸附试验(ELISA)法检测炎性因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达水平。将miR-155inhibitor转染入CFs,选择前述实验结果较为稳定的600μmol/L尿酸刺激CFs 24h,采用荧光定量PCR检测miR-155、IL-6、IL-1β、TNF-α、Ⅰ型胶原(Col-Ⅰ)、Ⅲ型胶原(Col-Ⅲ)的表达水平。结果与0μmol/L尿酸组相比较,400、600、800μmol/L尿酸组CFs的增殖和胶原含量增加(F=18.46、11.82,P<0.05),200、400、600、800μmol/L尿酸组CFs IL-6、IL-1β、TNF-α的表达增加(F=29.32～69.76,P<0.05)。miR-155inhibitor可明显抑制尿酸诱导的IL-6、IL-1β、TNF-α、Col-Ⅰ、Col-Ⅲ表达增加(F=72.74～275.32,P<0.05)。结论尿酸可以通过miR-155促进CFs的胶原合成及炎性分泌。更多还原

【Abstract】 Objective To investigate the effects of uric acid on collagen synthesis and inflammatory secretion in primary cultured neonatal rat cardiac fibroblasts(CFs)and the role of microRNA-155(miR-155)in the process. Methods The CFs of neonatal rats were isolated and treated with different concentrations of uric acid(0,200,400,600,and 800 μmol/L).The proliferation of CFs was evaluated by MTT assay,the collagen content was determined by hydroxyproline assay kit,and the expression of inflammatory factors interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)was determined by enzyme-linked immunosorbent assay.The CFs were then transfected with miR-155 inhibitor and stimulated with 600 μmol/L uric acid,which yielded stable results in the above test,for 24 h.The expression of miR-155,IL-6,IL-1β,TNF-α,collagen typeⅠ(Col-Ⅰ),and collagen type Ⅲ(Col-Ⅲ)was determined by quantitative real-time PCR. Results Compared with the group treated with 0μmol/L uric acid,the groups treated with 400,600,and 800 μmol/L uric acid had significantly increased proliferation and collagen content of CFs(F=18.46 and 11.82,P<0.05),and the groups treated with 200,400,600,and 800 μmol/L uric acid had significantly increased expression of IL-6,IL-1β,and TNF-αin CFs(F=29.32-69.76,P<0.05).However,the increased expression of IL-6,IL-1β,TNF-α,Col-Ⅰ,and Col-Ⅲ induced by uric acid was significantly inhibited by miR-155 inhibitor(F=72.74-275.32,P<0.05). Conclusion Uric acid acts through miR-155 to promote collagen synthesis and inflammatory secretion of CFs.更多还原