摘要
目的探讨体外培养人眼小梁细胞是否存在血红素氧合酶1(HO1)的表达及其抗氧化损伤作用。方法体外培养人眼小梁细胞,应用逆转录聚合酶链反应(RT PCR)检测小梁细胞中HO1mRNA的表达,免疫组织化学和Westernblot方法对HO1蛋白进行检测。向细胞培养液中加入H2O2,RT PCR和Westernblot检测小梁细胞HO1mRNA和蛋白的变化。分别预先加入HO1诱导剂氯化血红素(Hemin)和抑制剂锌原卟啉Ⅸ(ZnPPⅨ)1h后,通过MTT法检测小梁细胞在400μmol/LH2O2作用时的存活率。结果人眼小梁细胞存在HO1mRNA和蛋白。H2O2对小梁细胞HO1mRNA和蛋白的诱导呈浓度依赖性。Hemin可浓度依赖性地提高小梁细胞在H2O2作用时的存活率,HO1抑制剂ZnPPⅨ则浓度依赖性地降低小梁细胞在H2O2作用时的存活率。结论人眼小梁细胞存在HO1,并可被H2O2所诱导。HO1表达升高可提高小梁细胞抗氧化损伤的能力。
Objective To study the expression of heme oxygenase-1 （HO-1） in the cultured human trabecular meshwork cells （HTMCs） in vitro and the resistance against oxidant-induced injury. Methods HTMCs of the fourth generation were cultured in vitro. Reverse transcriptase-polymerase chain reaction （RT-PCR） was employed for detection of HO-1 mRNA in HTMCs. Immunohistochemical stain and Western blot were used to detect the expression of HO-1 protein. After H2O2 was added into the culture solution, the HO-1 mRNA levels were quantified by RT-PCR and the relative amounts of HO-1 protein measured by Western blot. After HTMCs were pretreated for 1h with HO-1 inducer heroin or HO-1 inhibitor zinc protoporphyrin IX （ZnPP-Ⅸ）, MTT assay was used to determine the survival rate of HTMCs in the presence of 400 mol/L H2O2. Results HO-1 was expressed in HTMCs. H2O2 induced HO-1 mRNA and protein in a dose-dependent manner. The survival rate of HTMCs in the presence of 400 mol/L H2O2 was increased in a dose-dependent manner by hemin, and was decreased in a dosedependent manner by ZnPP-Ⅸ. Conclusion HO-1 exists in the cultured HTMCs and can be induced by H2O2. The up-reguIation of HO-1 in HTMCs can increase the ability of HTMCs against oxidant-induced injury.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2005年第4期478-481,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
