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本文旨在研究硫化氢(hydrogen sulfide,H2S)对大鼠心脏成纤维细胞增殖的抑制作用。以原代培养新生大鼠心脏成纤维细胞(neonatal ratcardiac fibroblasts,NRCFs)为研究对象,用不同浓度血管紧张素II(angiotensin Ⅱ,Ang Ⅱ)或胎牛血清(fetalbovineserum,FBS)刺激NRCFs,建立NRCFs增殖模型。不同浓度硫氢化钠(NaHS,H2S的供体)处理该NRCFs增殖模型后,采用5’-溴-2’-脱氧尿嘧啶(5’-bromo-2’-deoxyuridine,BrdU)掺入法检测NRCFs增殖情况,用2’,7’-二氯荧光素乙酰乙酸盐(2’,7’-di-chlorofluorescein diacetate,DCFH-DA)荧光探针法检测细胞活性氧类(reactive oxygen species,ROS)水平。结果显示,较低浓度的NaHS(1×105mol/L)能促进FBS(2%、10%)对NRCFs的诱导增殖作用,但对Ang Ⅱ(1×107mol/L)所引起的NRCFs增殖的作用不明显,而较高浓度NaHS(5×105、1×104mol/L)则显著抑制10%FBS和Ang Ⅱ(1×107mol/L)对NRCFs的诱导增殖作用;1×104mol/LNaHS可显著降低Ang Ⅱ(1×107mol/L)诱导升高的细胞内ROS水平。以上结果表明,H2S在一定浓度范围内具有抑制NRCFs增殖的作用,其机制可能与H2S降低细胞内ROS有关。
This article aims to study the inhibitory effect of hydrogen sulfide (H2S) on the proliferation of rat cardiac fibroblasts. NRCFs were stimulated with different concentration of angiotensin Ⅱ (Ang Ⅱ) or fetal bovine serum (FBS) in primary cultured neonatal rat cardiac fibroblasts (NRCFs) to establish NRCFs Proliferation model. The proliferation of NRCFs was induced by different concentrations of sodium hydrosulfide (NaHS, donor of H2S), and then the proliferation of NRCFs was detected by using 5’- bromo-2’-deoxyuridine (BrdU) The reactive oxygen species (ROS) was detected by fluorescence probe with 2 ’, 7’-di-chlorofluorescein diacetate (DCFH-DA) Level. The results showed that NaHS at a low concentration (1 × 105mol / L) promoted the proliferation of NRCFs induced by FBS (2%, 10%), but had no effect on the proliferation of NRCFs induced by Ang Ⅱ (1 × 107mol / L) (5 × 105, 1 × 104mol / L) significantly inhibited the proliferation of NRCFs induced by 10% FBS and Ang Ⅱ (1 × 107mol / L), while the concentration of 1 × 104mol / Ang Ⅱ (1 × 107mol / L) induced elevated intracellular ROS levels. The above results show that H2S can inhibit the proliferation of NRCFs in a certain concentration range, and its mechanism may be related to the decrease of intracellular ROS by H2S.