新型钠磷转运蛋白抑制剂DZ1462在5/6肾切除高磷血症模型大鼠中的药效研究

doi:10.12300/j.issn.1674-5817.2021.138

摘要/Abstract

摘要：

目的探讨新型肠道钠磷转运蛋白小分子抑制剂DZ1462在大鼠5/6肾切除高磷血症动物模型中的降磷效果。方法首先随机选取156只Wistar大鼠，分为4组：第Ⅰ组6只为正常对照，饲喂正常饲料；第Ⅱ组60只，经过5/6肾切除后饲喂正常饲料；第Ⅲ组60只，经过5/6肾切除后饲喂高磷饲料；第Ⅳ组30只为假手术组，即只打开腹腔，不摘除肾脏，闭合伤口后饲喂高磷饲料。造模周期为10周。每两周检测各组大鼠血清磷含量，记录大鼠死亡数量，并进行HE染色观察肾脏病理变化，筛选高磷血症动物模型。选取符合入组标准的18只模型大鼠（均来源于第Ⅲ组），随机分为3组：模型对照组，记为G2组；DZ1462给药组（每次30 mg/kg，每日3次），记为G3组；商品化的降血磷药物碳酸司维拉姆片（Sevelamer）给药组（每次250 mg/kg，每日3次），记为G4组。每组6只，连续给药21 d。另外设正常对照组，记为G1组。运用无机磷检测试剂盒分析各组大鼠的血清磷水平变化。结果在手术建模后的第8周和第10周，5/6肾切除手术+高磷饲料饲喂的第Ⅲ组大鼠血清磷水平均明显高于正常对照第Ⅰ组（P＜0.01）；第Ⅲ组大鼠的肾脏出现了明显的肾小球硬化、肾小管萎缩、退化、间质炎症、纤维化和钙化，与人慢性肾病并发的高磷血症相似，提示肾性高磷血症动物模型建立成功。用药后，在各测量时间点上，DZ1462组大鼠的血清磷抑制率显著高于Sevelamer组（P＜0.05）。结论 DZ1462作为一种新型肠道钠磷转运蛋白小分子抑制剂，在大鼠高磷血症模型中能够有效抑制肠道磷离子的吸收，有望成为临床上治疗高磷血症的一种潜在有效药物。

关键词: 5/6肾切除手术, 高磷血症, 钠磷转运蛋白抑制剂, DZ1462, 大鼠

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

中图分类号:

Q95-33

卢晓, 张林, 季辉, 江善祥. 新型钠磷转运蛋白抑制剂DZ1462在5/6肾切除高磷血症模型大鼠中的药效研究[J]. 实验动物与比较医学, 2022, 42(3): 187-193.

Xiao LU, Lin ZHANG, Hui JI, Shanxiang JIANG. 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.

图/表 5

表1 各组大鼠在建模10周内的死亡率

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

图1 不同组别大鼠血清磷测试结果注：GⅠ，第Ⅰ组正常大鼠饲喂正常饲料；GⅡ，第Ⅱ组5/6肾切除大鼠饲喂正常饲料；GⅢ，第Ⅲ组5/6肾切除大鼠饲喂高磷饲料；GⅣ，第Ⅳ组假手术大鼠饲喂高磷饲料。**表示第Ⅰ组和第Ⅲ组在第8周和第10周时血清磷含量差异有极显著的统计学意义（P<0.01）。

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）.

图2 5/6肾切除手术模型大鼠的肾脏组织病理评估（HE染色）注：GⅠ为第Ⅰ组正常大鼠饲喂正常饲料，GⅢ为第Ⅲ组5/6肾切除高磷血症模型大鼠。上方图片放大倍数为100，下方图片放大倍数为200；实线箭头示间质炎症纤维化，虚线箭头示肾小球硬化。

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.

图 3 用药后不同组大鼠血清磷水平的比较注：G1为不手术+正常饲料喂养的正常对照大鼠；G2为5/6肾切除高磷血症模型对照组大鼠饲喂正常饲料；G3为5/6肾切除高磷血症模型大鼠经灌胃给予DZ1462 30 mg/kg并饲喂高磷饲料；G4为5/6肾切除高磷血症模型大鼠经灌胃给予盐酸酸司维拉姆（Sevelamer）250 mg/kg并饲喂高磷饲料。**表示G2组大鼠在用药第0、7、14、21天（即D0、D7、D14、D21）血清磷水平均与G1正常对照组显著增高（P < 0.01）。

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）.

《实验动物与比较医学》2022年征订启事

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