Efficacy of DZ1462, a Novel Sodium-phosphate Transporter Inhibitor, on 5/6 Nephrectomy-induced Hyperphosphatemia Model Rats

doi:10.12300/j.issn.1674-5817.2021.138

Abstract

Abstract:

Objective To study the efficacy of DZ1462, a novel sodium-phosphate transporter inhibitor, on rat hyperphosphatemia models established by 5/6 nephrectomy.Methods Totally 156 rats were randomly selected into four groups. Rats fed a normal diet were control group, named as group Ⅰ (n=6); rats fed a normal diet after 5/6 nephrectomy were named as group Ⅱ (n=60); rats fed a high phosphate diet after 5/6 nephrectomy were named as group Ⅲ (n=60); rats fed a high phosphate diet after sham surgery were named as group Ⅳ (n=30). The molding cycle was 10 weeks. Serum Pi was detected and the number of animal deaths was recorded every two weeks. Hematoxylin-eosin (HE) staining was performed to observe the change in kidney pathology, and to screen animal models with high phosphorus blood syndrome. Totally 18 model rats that met the inclusion criteria (all of group Ⅲ) were selected and randomly assigned to three groups: the model control group recorded as the G2 group; the DZ1462 administration group (30 mg/kg, tid, 21 d) recorded as the G3 group; the Sevelamer administration group (250 mg/kg, tid, 21 d) recorded as the G4 group. In addition, the normal control group was set as the G1 group. Serum phosphate levels were measured using a kit.Results In the 8^th and 10^th weeks, compared to group Ⅰ, serum phosphorus in group Ⅲ showed a significant difference (P < 0.01). The kidneys in group Ⅲ had obvious glomerular sclerosis, renal tubular atrophy, degeneration, interstitial inflammation, fibrosis, and calcification. Similarly to chronic kidney disease accompanied by hyperphosphatemia, the animal model was established successfully. At each time point, the serum phosphorus inhibition rate of the G3 group was significantly higher than that of the G4 group (P < 0.05).Conclusion DZ1462, as a novel small-molecule inhibitor of intestinal sodium and phosphorus transporter, can effectively inhibit intestinal phosphorus ion absorption in rat hyperphosphatemia model, and is expected to become a potential drug for the clinical treatment of hyperphosphatemia.

Key words: 5/6 Nephrectomy, Hyperphosphatemia, Sodium-phosphate transporter inhibitors, DZ1462, Rats

CLC Number:

Q95-33

Xiao LU,Lin ZHANG,Hui JI,et al. Efficacy of DZ1462, a Novel Sodium-phosphate Transporter Inhibitor, on 5/6 Nephrectomy-induced Hyperphosphatemia Model Rats[J]. Laboratory Animal and Comparative Medicine, 2022, 42(3): 187-193. DOI: 10.12300/j.issn.1674-5817.2021.138.

Figures/Tables 5

Table 1 Mortality of different group rats within 10 weeks after modeling

分组 Group	动物数量 Animal n	第1周 1^st Week		第3周 3^rd Week		第5周 5^th Week		第7周 7^th Week		第10周 10^th Week
分组 Group	动物数量 Animal n	No.	Rd%	No.	Rd%	No.	Rd%	No.	Rd%	No.	Rd%
Ⅰ	6	0	0	0	0	0	0	0	0	0	0
Ⅱ	60	6	10	3	15	0	15	6	25	5	30
Ⅲ	60	8	13	2	16	2	20	2	23	10	40
Ⅳ	30	0	0	1	3	1	6	0	6	0	6

Figure1 Serum phosphorus levels in different groups of ratsNote： GⅠ， normal rats with a normal diet （groupⅠ）；GⅡ， 5/6 nephrectomy rats with a normal diet （group Ⅱ）；GⅢ， 5/6 nephrectomy rats with a high Pi diet （group Ⅲ）；GⅣ， sham surgery rats with a high Pi diet （group Ⅳ）. **Represent serum Pi has a highly significant difference between Group Ⅰ and Group Ⅲ at the 8th and 10th week （P < 0.01）.

Figure 2 Pathological evaluation of kidney tissue in 5/6 nephrectomy model rats （HE staining）Note： GⅠ is normal rats with a normal diet （groupⅠ）， GⅢ is 5/6 nephrectomy rats with a high Pi diet （groupⅢ）. The magnification of the upper and the lower is 100 and 200， respectively. The solid arrow shows the interstitial fibrosis and inflammatory， and the dashed arrow shows glomerular sclerosis.

Figure 3 Comparison of serum phosphorus levels among different groups of rats after drug treatmentNote： G1 is normal rats with a normal diet； G2 is 5/6 nephrectomy and high diet model rats received a normal diet； G3 is 5/6 nephrectomy and high diet model rats received a high Pi diet and DZ1462 30 mg/kg； G4 is 5/6 nephrectomy and high diet model rats received a high Pi diet and Sevelamer 250 mg/kg. **Represent serum Pi in G2 is significantly higher than that in G1 on days 0， 7， 14， and 21 （P < 0.01）.

References 25

1	CHINYERE I R, MOUKABARY T, HUTCHINSON M D, et al. Progression of infarct-mediated arrhythmogenesis in a rodent model of heart failure[J]. Am J Physiol Heart Circ Physiol, 2021, 320(1): H108-H116. DOI:10.1152/ajpheart.00639. 2020 .
2	FOURNIER M L, CLÉMENT T, AUSSUDRE J, et al. Contusion rodent model of traumatic brain injury: controlled cortical impact[J]. Methods Mol Biol, 2021, 2193:49-65. DOI:10.1007/978-1-0716-0845-6_6 .
3	MARRA A. Animal models for drug development for MRSA[J]. Methods Mol Biol, 2020, 2069:253-266. DOI:10.1007/978-1-4939-9849-4_17 .
4	YOUNG F G. Growth and diabetes in normal animals treated with pituitary (anterior lobe) diabetogenic extract[J]. Biochem J, 1945, 39(5):515-536. DOI:10.1042/bj0390515 .
5	SARKAR D, AGRAWAL A, PAL D K. Clinical assessment of stabilisation of renal function after nephrectomy[J]. Urologia, 2021, 88(3):223-226. DOI:10.1177/0391560320987799 .
6	CHANTLER C, LIEBERMAN E, HOLLIDAY M A. A rat model for the study of growth failure in uremia[J]. Pediatr Res, 1974, 8(2):109-113. DOI:10.1203/00006450-197402000-00007 .
7	CZÉH B, SIMON M. Benefits of animal models to understand the pathophysiology of depressive disorders[J]. Prog Neuropsychopharmacol Biol Psychiatry, 2021, 106:110049. DOI:10.1016/j.pnpbp.2020.110049 .
8	PÉREZ-LÓPEZ L, BORONAT M, MELIÁN C, et al. Animal models and renal biomarkers of diabetic nephropathy[J]. Adv Exp Med Biol, 2021, 1307:521-551. DOI:10.1007/5584_2020_527 .
9	MCCARRON A, PARSONS D, DONNELLEY M. Animal and cell culture models for cystic fibrosis: which model is right for your application? [J]. Am J Pathol, 2021, 191(2):228-242. DOI:10.1016/j.ajpath.2020.10.017 .
10	ROBINSON N B, KRIEGER K, KHAN F M, et al. The Current state of animal models in research: a review[J]. Int J Surg, 2019, 72:9-13. DOI:10.1016/j.ijsu.2019.10.015 .
11	GUO W, WANG L Y, HAN X, et al. Effect of parathyroid hormone on intestinal mucosal sodium dependent phosphorus transporter[J]. Iran J Kidney Dis, 2021, 1(1):48-55.
12	ASKARI H, SEIFI B, KADKHODAEE M. Evaluation of renal-hepatic functional indices and blood pressure based on the progress of time in a rat model of chronic kidney disease[J]. Nephrourol Mon, 2016, 8(3): e37840. DOI:10.5812/numonthly. 37840 .
13	LEVEY A S, STEVENS L A, CORESH J. Conceptual model of CKD: applications and implications[J]. Am J Kidney Dis, 2009, 53(3 ): S4-S16. DOI:10.1053/j.ajkd.2008.07.048 .
14	HAN Y, LU J S, XU Y, et al. Rutin ameliorates renal fibrosis and proteinuria in 5/6-nephrectomized rats by anti-oxidation and inhibiting activation of TGFβ1-smad signaling[J]. Int J Clin Exp Pathol, 2015, 8(5):4725-4734.
15	GHOSH S S, MASSEY H D, KRIEG R, et al. Curcumin ameliorates renal failure in 5/6 nephrectomized rats: role of inflammation[J]. Am J Physiol Renal Physiol, 2009, 296(5): F1146-F1157. DOI:10.1152/ajprenal.90732.2008 .
16	JING W H, NUNES A C F, FARZANEH T, et al. Phosphate binder, ferric citrate, attenuates Anemia, renal dysfunction, oxidative stress, inflammation, and fibrosis in 5/6 nephrectomized CKD rats[J]. J Pharmacol Exp Ther, 2018, 367(1):129-137. DOI:10.1124/jpet.118.249961 .
17	王婷玉. 虫草保肾颗粒对5/6肾切除大鼠钙磷代谢的影响[D]. 哈尔滨: 黑龙江省中医药科学院, 2019.
	WANG T Y. Effect of Chongcao Baoshen Granules on calcium and phosphor metabolism with 5/6 nephrectomy rats[D]. Harbin: Heilongjiang College of Traditional Chinese Medicine, 2019.
18	BAO Y W, YUAN Y, CHEN J H, et al. Kidney disease models: tools to identify mechanisms and potential therapeutic targets[J]. Zool Res, 2018, 39(2):72-86. DOI:10.24272/j.issn.2095-8137.2017.055 .
19	RANI L, GAUTAM N K. Drosophila renal organ as a model for identification of targets and screening of potential therapeutic agents for diabetic nephropathy[J]. Curr Drug Targets, 2018, 19(16):1980-1990. DOI:10.2174/1389450119666180808114533 .
20	HAMZAOUI M, DJERADA Z, BRUNEL V, et al. 5/6 nephrectomy induces different renal, cardiac and vascular consequences in 129/Sv and C57BL/6JRj mice[J]. Sci Rep, 2020, 10(1):1524. DOI:10.1038/s41598-020-58393-w .
21	李鑫宇, 李丹, 赵学智. 一步法5/6肾切除大鼠加高磷饮食快速建立肾衰竭致血管钙化动物模型法[J]. 中国中西医结合肾病杂志, 2010, 11(10):862-864, 后插二. DOI:10.3969/j.issn.1009-587X.2010.10.006 .
	LI X Y, LI D, ZHAO X Z. One-step approach of 5/6 nephrectomy plus high phosphorus diet to rapidly establish rat models of vessel calcification induced by renal failure[J]. Chinese Journal of Integrated Traditional and Western Nephrology, 2010, 11(10):862-864. DOI:10.3969/j.issn.1009-587X.2010.10.006 .
22	RADLOFF J, LATIC N, PFEIFFENBERGER U, et al. A phosphate and calcium-enriched diet promotes progression of 5/6-nephrectomy-induced chronic kidney disease in C57BL/6 mice[J]. Sci Rep, 2021, 11(1):14868. DOI:10.1038/s41598-021-94264-8 .
23	TSAO C W, HSU Y J, CHANG T C, et al. A high phosphorus diet impairs testicular function and spermatogenesis in male mice with chronic kidney disease[J]. Nutrients, 2020, 12(9):2624. DOI:10.3390/nu12092624 .
24	LARSSON T E, KAMEOKA C, NAKAJO I, et al. NPT-Ⅱb inhibition does not improve hyperphosphatemia in CKD[J]. Kidney Int Rep, 2017, 3(1):73-80. DOI:10.1016/j.ekir.2017.08.003 .
25	MARUYAMA S, MARBURY T C, CONNAIRE J, et al. NaPi-Ⅱb inhibition for hyperphosphatemia in CKD hemodialysis patients[J]. Kidney Int Rep, 2020, 6(3):675-684. DOI:10.1016/j.ekir.2020.12.017 .

[1]	QIN Chao, LI Shuangxing, ZHAO Tingting, JIANG Chenchen, ZHAO Jing, YANG Yanwei, LIN Zhi, WANG Sanlong, WEN Hairuo. Study on the 90-day Feeding Experimental Background Data of SD Rats for Drug Safety Evaluation [J]. Laboratory Animal and Comparative Medicine, 2025, 45(4): 439-448.
[2]	LIU Liyu, JI Bo, LIU Xiaoxuan, FANG Yang, ZHANG Ling, GUO Tingting, QUAN Ye, LI Hewen, LIU Yitian. Exploration of Rat Fetal Lung Tissue Fixation Methods [J]. Laboratory Animal and Comparative Medicine, 2025, 45(4): 432-438.
[3]	LIU Zhiwei, YANG Ran, LIAN Hao, ZHANG Yu, JIN Lilun. Cartilage Protection and Anti-Inflammatory Effects of Fraxetin on Monosodium Iodoacetate-Induced Rat Model of Osteoarthritis [J]. Laboratory Animal and Comparative Medicine, 2025, 45(3): 259-268.
[4]	JIANG Meng, HAO Shulan, TONG Liguo, ZHONG Qiming, GAO Zhenfei, WANG Yonghui, WANG Xixing, JI Haijie. Dynamic Evaluation of Vinorelbine-Induced Phlebitis of Dorsalis Pedis Vein in a Rat Model [J]. Laboratory Animal and Comparative Medicine, 2025, 45(3): 251-258.
[5]	PAN Yicong, JIANG Wenhong, HU Ming, QIN Xiao. Optimization of Surgical Procedure and Efficacy Evaluation of Aortic Calcification Model in Rats with Chronic Kidney Disease [J]. Laboratory Animal and Comparative Medicine, 2025, 45(3): 279-289.
[6]	LIAN Hui, JIANG Yanling, LIU Jia, ZHANG Yuli, XIE Wei, XUE Xiaoou, LI Jian. Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding [J]. Laboratory Animal and Comparative Medicine, 2025, 45(2): 130-146.
[7]	YIN Yulian, MA Lina, TU Siyuan, CHEN Ling, YE Meina, CHEN Hongfeng. Establishment and Evaluation of a Rat Model of Non-Puerperal Mastitis [J]. Laboratory Animal and Comparative Medicine, 2024, 44(6): 587-596.
[8]	YANG Jin, YU Shiya, LIN Nan, FANG Yongchao, ZHAO Hu, QIU Jinwei, LIN Hongming, CHEN Huiyan, WANG Yu, WU Weihang. Effect of Modified Duodenal Exclusion Surgery on Glucose Metabolism in Rats with Type 2 Diabetes Mellitus [J]. Laboratory Animal and Comparative Medicine, 2024, 44(5): 523-530.
[9]	QI Longju, CHEN Shiyuan, LIAO Zehua, SHI Yuanhu, SUN Yuyu, WANG Qinghua. Transcriptomic Analysis of Menstrual Blood-Derived Stem Cells Transplantation Combined with Exercise Training in Promoting Spinal Cord Injury Recovery in Rats [J]. Laboratory Animal and Comparative Medicine, 2024, 44(5): 531-542.
[10]	ZHANG Naiqun, YUAN Piaopiao, CAO Linrong, YING Na, YANG Taotao. Application of PNR Detection in the Diagnosis and Drug-efficacy Evaluation of Diabetic Kidney Disease in Rats [J]. Laboratory Animal and Comparative Medicine, 2024, 44(5): 543-549.
[11]	ZHENG Yiqing, DENG Yasheng, FAN Yanping, LIANG Tianwei, HUANG Hui, LIU Yonghui, NI Zhaobing, LIN Jiang. Application Analysis of Animal Models for Pelvic Inflammatory Disease Based on Data Mining [J]. Laboratory Animal and Comparative Medicine, 2024, 44(4): 405-418.
[12]	XIAO Pan, WANG Hongyi, LU Lu, ZHANG Mei, CHEN Keming, SHEN Dongshuai, NIU Tingxian. Screening of Hypoxia-Sensitive and Hypoxia-Tolerant Wistar Rats and Preliminary Exploration of Hypoxia Sensitivity in Their G₁Generation [J]. Laboratory Animal and Comparative Medicine, 2024, 44(4): 374-383.
[13]	Xiaoyu ZHU, Hantao YUAN, Sibo LI. MicroRNA-887-3p Inhibited MDM4 Expression and Proliferation but Promoted Apoptosis of Intervertebral Disc Annulus Fibrosus Cells in Rats [J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 270-278.
[14]	Jinhua HU, Jingjie HAN, Min JIN, Bin HU, Yuefen LOU. Effects of Puerarin on Bone Density in Rats and Mice: A Meta-analysis [J]. Laboratory Animal and Comparative Medicine, 2024, 44(2): 149-161.
[15]	Liya ZHAO, Liju NI, Caiqin ZHANG, Jianping TANG, Yangzheng YAO, Yanyan NIE, Xiaoxue GU, Ying ZHAO. Establishing a Genetic Detection Protocol of Single Nucleotide Polymorphisms Panels in Inbred Rats Based on Multiplex PCR-LDR [J]. Laboratory Animal and Comparative Medicine, 2023, 43(5): 548-558.