低氧诱导因子-1α对牦牛肾小管上皮细胞中自噬相关因子表达的调控

HIF-1αregulates the expression of autophagy related factors in yak renal tubular epithelial cells

本研究旨在探究长期高海拔低氧环境对牦牛肾小管上皮细胞(RTECs)中自噬相关因子表达的影响。通过应用免疫组织化学、免疫荧光、qRT-PCR、Western-blot试验,比较了成年牦牛和黄牛肾脏中HIF-1α的分布和表达情况。免疫组织化学和免疫荧光染色结果显示,HIF-1α在成年牦牛和黄牛肾脏组织中均呈阳性表达,主要分布在肾小管,集合管上皮细胞、肾小管上皮细胞中。Western-blot和qRT-PCR结果表明,成年牦牛肾脏中HIF-1α的表达显著高于黄牛(P<0.0001)。使用原代培养的牦牛RTECs,通过MDC自噬染色等试验方法,研究了HIF-1α药物抑制和过表达对自噬相关因子表达作用。使用药物DMOG诱导RTECs中的HIF-1α的表达上调时,BNIP3、Beclin-1蛋白和m RNA的表达量升高(P<0.0001),而caspase-3降低(P<0.0001);使用药物LW6诱导HIF-1α的表达下调时,BNIP3、Beclin-1蛋白和m RNA的表达量降低(P<0.001),而caspase-3(P<0.0001)升高。研究结果表明HIF-1α是调控牦牛肾脏中自噬相关因子表达的重要基因。

This study aimed to investigate the impact of long-term high-altitude hypoxic environments on the expression of autophagy-related factors in renal tubular epithelial cells(RTECs)of yaks.Through the application of immunohistochemistry,immunofluorescence,q RT-PCR,Western-blot,and other experiments,the distribution and expression of HIF-1αin the kidneys of adult yaks and domestic cattle were compared.Immunohistochemical and immunofluorescence staining results demonstrated positive expression of HIF-1αin kidney tissues of adult yaks and cattle,predominantly distributed in renal tubules,collecting duct epithelial cells,and renal tubular epithelial cells.WB and q RT-PCR results indicated significantly higher expression of HIF-1αin the kidneys of adult yaks compared to cattle(P<0.0001).Using primary cultured yak RTECs and experimental methods such as MDC autophagy staining,the regulatory effects of drug inhibition and overexpression of HIF-1αon the expression of autophagy and apoptosis-related factors were investigated.When inducing the upregulation of HIF-1αexpression in RTECs using the drug DMOG,the expression levels of BNIP3,Beclin-1 protein,and m RNA increased(P<0.0001),while caspase-3 decreased(P<0.0001);conversely,when inducing the downregulation of HIF-1αexpression using the drug LW6,the expression levels of BNIP3,Beclin-1 protein,and m RNA decreased(P<0.001),while caspase-3 increased(P<0.0001).The results of the study indicate that HIF-1αis an important gene regulating the expression of autophagy-related factors in yak kidneys.