雄激素通过HIF-1α/BNIP3信号通路介导肾小管上皮细胞凋亡在草酸钙肾结石形成中的机制研究

The incidence of kidney stones is very high, seriously affecting the quality of life of patients. Calcium oxalate kidney stone is the most common type. Previous studies indicated that apoptosis of renal tubular epithelia cell (TEC) and androgen might play a key role in the development of nephrolithiasis. However, mechanisms are still unknown. HK-2 cells were transfected with Lentiviruses expressing AR shRNA targeting AR, and cells under different treatments were collected for microarray analysis. BNIP3 were identified in most differentially expressed genes and were regulated by androgen.The use of bioinformatics were proved that HIF-1α may be brige molecule between androgen and BNIP3. In addition,Assays were performed to prove HIF-1α increased BNIP3 expression by combination with its promoter region.These results indicate that testosterone induce renal TEC cell apoptosis by activating HIF-1α/BNIP3 pathway.This research program is intended to explore the effects and the underlying mechanism of testosterone-induced renal TEC cell apoptosis on HIF-1α/BNIP3 pathway in vitro and in vivo by using co-immunoprecipitation and point mutation, dual luciferase reporter gene,protein Mass Spectrometry and androgen receptor knockout mouse model. We will carry out the following researches. (1)The role of BNIP3 in androgen-induced apoptosis; (2)The mechanism of HIF-1α on Regulation of BNIP3 Expression. Therefore providing a new theory for the prevention and treatment of calcium oxalate and kidney stones.

肾结石发病率高,严重影响患者的生活质量,其中草酸钙结石最常见.肾小管上皮细胞凋亡是结石形成的中心环节,雄激素可通过雄激素受体（AR）促进肾小管上皮细胞凋亡,但具体机制不明.前期我们通过基因芯片筛查过表达及干扰AR后肾小管上皮细胞基因表达谱变化,结果发现BCL2和腺病毒E1B19kDa相互作用蛋白3（BNIP3）表达差异明显,且受雄激素正向调控.进一步利用生物信息学预测HIF-1α可能是雄激素与BNIP3作用的桥梁分子,我们前期研究证实HIF-1α能够增强BNIP3启动子活性并上调其表达.据此推测:雄激素可能通过HIF-1α/BNIP3信号通路促进肾小管上皮细胞凋亡.本研究拟从体内外利用染色质免疫共沉淀及点突变,双荧光素酶报告基因,蛋白质谱,雄激素受体敲除小鼠模型等方法阐明①BNIP3在雄激素诱导细胞凋亡中的作用②HIF-1α调控BNIP3表达的机制,从而为草酸钙肾结石的防治提供新的理论依据

肾结石疾病在全世界范围都很常见，其发病率及复发率均高，常可引起肾功能的损害，造成极大危害。近年来，随着结石微创治疗技术的日渐成熟，已经基本能解决所有部位的结石。然而关于结石形成机制目前仍不明，因此，探究其发病机制，对于预防和治疗肾结石有重要意义。经典的结石形成理论认为肾小管上皮细胞凋亡及损伤是肾结石形成的中心环节。本研究利用体内外实验研究了BNIP3介导肾小管上皮细胞凋亡的机制，并且探究EZH2抑制剂能减轻高草酸刺激环境下诱导的肾损伤。从而为草酸钙肾结石的防治提供新的理论依据。相关研究发表SCI论文一篇（Xiaomin Gao#, Yonghan Peng#, Ziyu Fang#, et al. Inhibition of EZH2 ameliorates hyperoxaluria-induced kidney injury through the JNK/FoxO3a pathway, Life Sciences (2021) IF：5.03）。