Yang Huan, Male Musa, Li Yang, Wang Ning, Zhao Chenming, Jin Shan, Hu Juncheng, Chen Zhiqiang, Ye Zhangqun, Xu Hua
Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095# Jie Fang Avenue, Wuhan, 430030, China.
Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
BMC Nephrol. 2018 Jul 6;19(1):167. doi: 10.1186/s12882-018-0980-8.
Substrate reduction therapy with analogs reduces the accumulation of substrates by inhibiting the metabolic pathways involved in their biosynthesis, providing new treatment options for patients with primary hyperoxalurias (PHs) that often progress to end-stage renal disease (ESRD). This research aims to evaluate the inhibition efficacy of Hydroxy-L-proline (HYP) analogs against calcium oxalate (CaOx) crystal formation in the Drosophila Melanogaster (D. Melanogaster) by comparing them with Pyridoxine (Vitamin B6).
Three stocks of Drosophila Melanogaster (W, CG3926 RNAi, and Act5C-GAL4/CyO) were utilized. Two stocks (CG3926 RNAi and Act5C-GAL4 /CyO) were crossed to generate the Act5C > dAGXT RNAi recombinant line (F generation) of D. Melanogaster which was used to compare the efficacy of Hydroxy-L-proline (HYP) analogs inhibiting CaOx crystal formation with Vitamin B as the traditional therapy for primary hyperoxaluria.
Nephrolithiasis model was successfully constructed by downregulating the function of the dAGXT gene in D. Melanogaster (P-Value = 0.0045). Furthermore, the efficacy of Hydroxy-L-proline (HYP) analogs against CaOx crystal formation was demonstrated in vivo using D. Melanogaster model; the results showed that these L-Proline analogs were better in inhibiting stone formation at very low concentrations than Vitamin B (IC = 0.6 and 1.8% for standard and dietary salt growth medium respectively) compared to N-acetyl-L-Hydroxyproline (IC = 0.1% for both standard and dietary salt growth medium) and Baclofen (IC = 0.06 and 0.1% for standard and dietary salt growth medium respectively). Analysis of variance (ANOVA) also showed that Hydroxy-L-proline (HYP) analogs were better alternatives for CaOx inhibition at very low concentration especially when both genetics and environmental factors are intertwined (p < 0.0008) for the dietary salt growth medium and (P < 0.063) for standard growth medium.
Addition of Hydroxy-L-Proline analogs to growth medium resulted in the reduction of CaOx crystals formation. These analogs show promise as potential inhibitors for oxalate reduction in Primary Hyperoxaluria.
使用类似物进行底物还原疗法通过抑制参与其生物合成的代谢途径来减少底物的积累，为常进展至终末期肾病（ESRD）的原发性高草酸尿症（PHs）患者提供了新的治疗选择。本研究旨在通过将羟基-L-脯氨酸（HYP）类似物与吡哆醇（维生素B6）进行比较，评估其对果蝇中草酸钙（CaOx）晶体形成的抑制效果。
使用了三株果蝇（W、CG3926 RNAi和Act5C-GAL4/CyO）。将两株（CG3926 RNAi和Act5C-GAL4 /CyO）进行杂交，以产生果蝇的Act5C > dAGXT RNAi重组品系（F代），该品系用于将羟基-L-脯氨酸（HYP）类似物抑制CaOx晶体形成的效果与作为原发性高草酸尿症传统疗法的维生素B进行比较。
通过下调果蝇中dAGXT基因的功能成功构建了肾结石模型（P值 = 0.0045）。此外，使用果蝇模型在体内证明了羟基-L-脯氨酸（HYP）类似物对CaOx晶体形成的抑制效果；结果表明，与N-乙酰-L-羟基脯氨酸（标准和膳食盐生长培养基的IC均为0.1%）和巴氯芬（标准和膳食盐生长培养基的IC分别为0.06%和0.1%）相比，这些L-脯氨酸类似物在非常低的浓度下抑制结石形成的效果优于维生素B（标准和膳食盐生长培养基的IC分别为0.6%和1.8%）。方差分析（ANOVA）还表明，羟基-L-脯氨酸（HYP）类似物在非常低的浓度下是抑制CaOx的更好选择，特别是当遗传和环境因素相互交织时（膳食盐生长培养基的p < 0.0008，标准生长培养基的P < 0.063）。
在生长培养基中添加羟基-L-脯氨酸类似物可减少CaOx晶体的形成。这些类似物有望成为原发性高草酸尿症中草酸盐减少的潜在抑制剂。
