摘要
目的:SLC34a2基因编码的钠依赖性磷转运蛋白Na Pi-IIb在磷的吸收中起重要作用,减少CKD患者磷的肠道吸收可能是治疗其高磷血症的潜在方案。本研究旨在探索萝卜硫素(sulforaphane,SFN)能否通过拮抗孕烷X受体(pregnane X receptor,PXR)抑制人源性结肠癌细胞中Na Pi-IIb表达,为高磷血症寻找新治疗靶点。方法:RT-qPCR检测3种结肠癌细胞系HT29、LOVO及SW480中PXR及SLC34a2 mRNA表达量;MTT法确定SFN及利福平(Rifampicin,RFP)的适宜作用浓度和时间;RT-qPCR和Westernblot分别检测药物处理后细胞系SLC34a2 mRNA和NaPi-IIb的表达情况;观察探讨SFN、PXR及Na Pi-IIb三者之间的关系。结果:(1)10μmol/L和20μmol/L SFN均可抑制LOVO细胞中SLC34a2 mRNA及NaPi-IIb表达,且后者的抑制效果更强。(2)10μmol/L SFN可降低由10μmol/L RFP诱导的SLC34a2 mRNA和Na Pi-IIb高表达。结论:SFN对SLC34a2 mRNA及NaPi-IIb表达存在确切抑制作用;SFN可竞争性抑制RFP激活PXR引起的Na Pi-IIb升高,表明SFN对NaPi-IIb的抑制作用可通过SFN-/-PXR-NaPi-IIb通路实现。
Objective ： NaPi - IIb sodium - dependent phosphate transporter protein, which is encoded by SLC34a2 gene, plays a dominant role in intestinal phosphorus absorption. The reduction of intestinal phosphorus absorption is a potential treatment tot hyperphosphatemia in CKD patients. This paper aimed to investigate whether sulforaphane （SFN） could reduce the expression of NaPi - lib through antagonizing pregnane X receptor （PXR）. Methods： RT - qPCR was conducted to examine the expression levels of PXR and SLC34a2 in three colon cancer cell lines, HT29 ,LOVO and SW480. MTT assay was conducted to determine the proper dose and time for SFN and Rifampicin（RFP） treatment. Then, RT - qPCR and westernblot were conducted to examine the expression level of SLC34a2 mRNA and NaPi - lib, respectively. Furthermore, the relationship among SFN, PXR and NaPi - Ilb was revealed. Re- sults：（ 1 ） SFN with concentration levels of 10 p, mol/L and 20 μmol/L significantly reduced the expression of SLC34a2 mRNA anti NaPi- lib in LOVO cells. Moreover, SFN with higher concentration level showed stronger suppresion effects. （2） 10 μmol/L SFN attenuated the increase of SLC34a2 mRNA and NaPi - lib induced by 10 μmol/L RFP. Conclusion： SFN carl reduce the expression of NaPi - IIb and this effect is realized by a potential signal pathway SFN - / - PXR - NaPi -IIb.
出处
《中国中西医结合肾病杂志》
2016年第7期578-581,共4页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
陕西省社会发展科技攻关项目(No.2013K12-040-01)
