PNR检测在糖尿病肾脏疾病模型大鼠诊断及药效评价中的应用

doi:10.12300/j.issn.1674-5817.2024.044

实验动物与比较医学 ›› 2024, Vol. 44 ›› Issue (5): 543-549.DOI: 10.12300/j.issn.1674-5817.2024.044

PNR检测在糖尿病肾脏疾病模型大鼠诊断及药效评价中的应用

张乃群¹(), 袁飘漂¹, 曹琳茸¹, 应娜¹, 杨涛涛²()()

^1.余姚市第四人民医院, 余姚 315470
^2.宁波市医疗中心李惠利医院, 宁波 315000

收稿日期:2024-03-13 修回日期:2024-08-04 出版日期:2024-11-06 发布日期:2024-10-25
通讯作者: 杨涛涛（1989—），男，硕士，主管中药师，从事医院药学和药理学研究。E-mail： 843397827@qq.com。ORCID： 0009-0000-7268-9685
作者简介:张乃群（1979—），女，本科，副主任药师，主要从事医院药学和药理学研究。E-mail： 326943179@qq.com
基金资助:
余姚市卫生健康科技计划项目“基于U-PNR检测探究缬沙坦和卡托普利对DKD蛋白尿一级预防作用研究”(2022YYB13);浙江省药学会医院药学专项科研资助项目“基于尿PNR检测的DN早期诊断及缬沙坦的干预作用研究”(2017ZYY24)

Application of PNR Detection in the Diagnosis and Drug-efficacy Evaluation of Diabetic Kidney Disease in Rats

ZHANG Naiqun¹(), YUAN Piaopiao¹, CAO Linrong¹, YING Na¹, YANG Taotao²()()

^1.The No. 4 People's Hospital of Yuyao, Yuyao 315470, China
^2.Ningbo Medical Center Lihuili Hospital, Ningbo 315000, China

Received:2024-03-13 Revised:2024-08-04 Published:2024-10-25 Online:2024-11-06
Contact: YANG Taotao (ORCID: 0009-0000-7268-9685), E-mail: 843397827@qq.com

摘要/Abstract

摘要：

目的通过监测肾小球足细胞相关分子podocin和nephrin的mRNA比值（podocin mRNA： nephrin mRNA ratio，PNR）在大鼠糖尿病肾脏疾病（diabetic kidney disease，DKD）早期模型中的变化，并将其与尿白蛋白-肌酐比值（urinary albumin to creatinine ratio，U-ACR）对比，评价PNR作为DKD早期诊断指标的可行性；并比较缬沙坦和福辛普利钠对DKD的早期干预作用。方法采用尾静脉注射链脲霉素（streptozotocin，STZ）（60 mg/kg）建立DKD大鼠模型，监测对比PNR与U-ACR的早期变化，适时给予缬沙坦和福辛普利钠干预；采用苏木精-伊红（hematoxylin and eosin staining，HE）染色观察肾小球结构形态，透射电子显微镜观察肾小球足细胞超微结构。结果造模后PNR在第9天达临界值（≥1），较U-ACR在第15天达临界值（≥30 mg/g）更早；造模后第9天给予缬沙坦和福辛普利钠干预，发现其对U-ACR均有显著降低作用（P＜0.05），且缬沙坦低剂量优于高剂量（P＞0.05），而福辛普利钠高剂量优于低剂量（P＞0.05）；低剂量的缬沙坦和福辛普利钠均对PNR有明显降低作用（P＜0.05），高剂量未见明显影响；缬沙坦和福辛普利钠干预对肾小球结构及足细胞排布均具有一定的改善和维持作用。结论 PNR较U-ACR更早变化，对大鼠DKD早期诊断具有一定的提示作用，缬沙坦和福辛普利钠早期干预对大鼠DKD有一定的治疗作用。

关键词: PNR, 尿白蛋白-肌酐比值, 糖尿病肾脏疾病, 蛋白尿预防, 大鼠

Abstract:

Objective This study aims to monitor the mRNA ratio of podocin to nephrin (PNR) in glomerular podocytes of early diabetic kidney disease (DKD) rat models. The feasibility of using PNR as an early diagnostic indicator for DKD was evaluated by comparing it with the urinary albumin-to-creatinine ratio (U-ACR). Additionally, the early intervention effects of valsartan and fosinopril sodium on DKD were compared. Methods The DKD rat model was established by caudal intravenous injection of streptozotocin (STZ) at a dosage of 60 mg/kg. The early changes in PNR and U-ACR were monitored and compared, followed by timely intervention with valsartan and fosinopril sodium. Hematoxylin and eosin staining (HE) was used to observe glomerular structure, while transmission electron microscopy examined the ultrastructure of glomerular podocytes. Results PNR reached the critical value(≥1) on day 9 after modeling, earlier than U-ACR, which reached the critical value(≥30 mg/g) on day 15. Intervention with valsartan and fosinopril sodium on day 9 after modeling significantly reduced U-ACR (P < 0.05), with low-dose valsartan showing better results than high-dose (P>0.05), while high-dose fosinopril sodium outperformed low-dose (P>0.05). Both low doses of valsartan and fosinopril sodium significantly reduced PNR (P<0.05), with no significant effect observed for high doses. The interventions with valsartan and fosinopril sodium improved and maintained glomerular structure and podocyte arrangement. Conclusion PNR changes earlier than U-ACR, indicating its potential as an early diagnostic marker for DKD in rats. Early intervention with valsartan and fosinopril sodium demonstrates a therapeutic effect on DKD in rats.

Key words: PNR, Urinary albumin-to-creatinine ratio (U-ACR), Diabetic kidney disease, Proteinuria prevention, Rats

中图分类号:

R587.1

张乃群,袁飘漂,曹琳茸,等. PNR检测在糖尿病肾脏疾病模型大鼠诊断及药效评价中的应用[J]. 实验动物与比较医学, 2024, 44(5): 543-549. DOI: 10.12300/j.issn.1674-5817.2024.044.

ZHANG Naiqun,YUAN Piaopiao,CAO Linrong,et al. 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. DOI: 10.12300/j.issn.1674-5817.2024.044.

图/表 3

表1 STZ造模后大鼠U-ACR和PNR值的变化 (xˉ±s, n=3)

Table 1 Changes of U-ACR and PNR values in rats after STZ modeling

造模后时间 Time after molding/d	U-ACR/(mg·g^-1)		PNR
造模后时间 Time after molding/d	正常组 Normal	模型组 Model	正常组 Normal	模型组 Model
3	12.69±3.69	ND	0.85±0.40	0.39±0.19
5	13.29±4.05	ND	0.88±0.31	0.64±0.45
7	13.02±3.7	ND	0.86±0.37	0.66±0.10
9	12.14±3.27	9.45±6.23	0.91±0.27	1.26±0.11
11	11.96±3.21	18.19±8.75	0.90±0.25	1.70±0.65
13	13.88±4.02	ND	0.89±0.30	1.63±0.63
15	12.73±3.98	72.59±19.51	0.87±0.38	1.25±0.17
17	13.01±2.98	77.68±11.16	0.91±0.42	2.54±0.20

表2 药物干预对糖尿病模型大鼠U-ACR和PNR的影响 (xˉ±s, n=10)

Table 2 Effects of drug intervention on U-ACR and PNR in diabetic model rats

组别 Group	U-ACR/(mg·g^-1)		PNR
组别 Group	给药前 0 week	给药4周 4 weeks	给药前 0 week	给药4周 4 weeks
正常对照 Normal control (0.9%NS)	11.69±3.15	11.45±3.25	0.92±0.21	0.89±0.17
模型对照 Model control (0.9%NS)	17.96±6.45	896.12±62.89 ^**	1.24±0.11	1.64±0.08 ^**
缬沙坦低剂量 Valsartan 25 mg/kg	18.08±7.03	408.99±78.03 ^△△	1.16±0.14	0.53±0.13 ^△△
缬沙坦高剂量 Valsartan 50 mg/kg	18.86±6.90	480.97±94.34 ^△△	1.14±0.24	1.28±0.44
福辛普利钠低剂量 Fosinopril 30 mg/kg	18.73±6.82	314.93±67.81 ^△△	1.26±0.22	0.92±0.28 ^△△
福辛普利钠高剂量 Fosinopril 60 mg/kg	17.86±5.92	236.61±43.55 ^△△	1.17±0.21	2.14±0.29

图1 各组大鼠肾脏组织的HE染色（A～F，×400）和透射电子显微镜观察（A'～F'，×5 000）注：A、A' 为正常对照组（0.9% NaCl）；B、B' 为模型对照组（0.9% NaCl）；C、C' 为缬沙坦低剂量组（Valsartan 25 mg/kg）；D、D' 为缬沙坦高剂量组（Valsartan 50 mg/kg）；E、E' 为福辛普利钠低剂量组（Fosinopril 30 mg/kg）；F、F' 为福辛普利钠高剂量组（Fosinopril 60 mg/kg）。B图中，箭头①为肾小球形态轮廓，箭头②为系膜区基质沉积；B’图中，箭头①为足细胞基底膜弥漫性不规则增厚，箭头②③为足细胞“拉链状”结构布局破损、模糊不成型。

Figure 1 HE staining (A-F， ×400) and transmission electron microscopy (A'-F'， ×5 000) of renal tissues in each group of ratsNote：A and A’ represent the normal control group (0.9% NaCl), B and B' represent the model control group (0.9% NaCl), C and C' represent the low-dose Valsartan group (Valsartan 25 mg/kg), and D and D' represent the high-dose Valsartan group (Valsartan 50 mg/kg). E and E' represent the low-dose fosinopril sodium group (Fosinopril 30 mg/kg), and F and F’represent the high-dose fosinopril sodium group (Fosinopril 60 mg/kg). In fig. B, the arrow ① indicates the glomerular contour, the arrow ② indicates the mesangial matrix deposition. In fig. B', the arrow ① indicates the diffuse irregular thickening of the podocyte membrane, while the arrows ② and ③ indicate damage and fuzziness of the “zipper” layout of podocytes.

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PNR检测在糖尿病肾脏疾病模型大鼠诊断及药效评价中的应用

Application of PNR Detection in the Diagnosis and Drug-efficacy Evaluation of Diabetic Kidney Disease in Rats

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