Department of Basic Medicine, Anyang Vocational and Technical College, Anyang Henan 455000.
Second Department of Pediatrics, First Affiliated Hospital of Xinxiang Medical College, Weihui Henan 453100.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2022 Jan 28;47(1):26-34. doi: 10.11817/j.issn.1672-7347.2022.201001.
Nephrotic syndrome is a common disease of the urinary system. The aim of this study is to explore the effect of astragalus polysaccharides (APS) on multidrug resistance gene 1 (MDR1) and P-glycoprotein 170 (P-gp170) in adriamycin nephropathy rats and the underlying mechanisms.
A total of 72 male Wistar rats were divided into a control group, a model group, an APS low-dose group, an APS high-dose group, an APS+micro RNA (miR)-16 antagomir group and an APS+miR-16 antagomir control group, with 12 rats in each group. Urine protein (UP) was detected by urine analyzer, and serum cholesterol (CHOL), albumin (ALB), blood urea nitrogen (BUN), and creatinine (SCr) were detected by automatic biochemical analyzer; serum interleukin-6 (IL-6), IL-1β, tumor necrosis factor α (TNF-α) levels were detected by ELISA kit; the morphological changes of kidney tissues were observed by HE staining; the levels of miR-16 and MDR1 mRNA in kidney tissues were detected by real-time RT-PCR; the expression levels of NF-κB p65, p-NF-κB p65, and P-gp170 protein in kidney tissues were detected by Western blotting; and dual luciferase was used to verify the relationship between miR-16 and NF-κB.
The renal tissue structure of rats in the control group was normal without inflammatory cell infiltration. The renal glomeruli of rats in the model group were mildly congested, capillary stenosis or occlusion, and inflammatory cell infiltration was obvious. The rats in the low-dose and high-dose APS groups had no obvious glomerular congestion, the proliferation of mesangial cells was significantly reduced, and the inflammatory cells were reduced. Compared with the high-dose APS group and the APS+miR-16 antagomir control group, there were more severe renal tissue structure damages in the APS + miR-16 antagomir group. Compared with the control group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1β, TNF-α, and MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 in the model group were significantly increased (all <0.05); the levels of ALB and miR-16 were significantly decreased (both <0.05). Compared with the model group, the levels of UP, CHOL, BUN, SCr, IL-6, IL-1β, TNF-α, and MDR1 mRNA, and the protein levels of pNF-κB p65 and P-gp170 in the low-dose and high-dose APS groups were significant decreased (all <0.05); and the levels of ALB and miR-16 were significantly increased (both <0.05). Compared with APS+miR-16 antagomir control group, the UP, CHOL, BUN, SCr, IL-6, IL-1β, and TNF-α levels, MDR1 mRNA, and the protein levels of p-NF-κB p65 and P-gp170 were significantly increased (all <0.05). The levels of ALB and miR-16 were significantly decreased in the APS+miR-16 antagomir group compared with the APS+miR-16 antagomir control group (both <0.05).
APS can regulate the miR-16/NF-κB signaling pathway, thereby affecting the levels of MDR1 and P-gp170, and reducing the inflammation in the kidney tissues in the adriamycin nephropathy rats.
肾病综合征是一种常见的泌尿系统疾病。本研究旨在探讨黄芪多糖（APS）对阿霉素肾病大鼠多药耐药基因 1（MDR1）和 P-糖蛋白 170（P-gp170）的影响及其机制。
将 72 只雄性 Wistar 大鼠随机分为对照组、模型组、APS 低剂量组、APS 高剂量组、APS+微小 RNA（miR）-16 反义寡核苷酸组和 APS+miR-16 反义寡核苷酸对照组，每组 12 只。用尿分析仪检测尿蛋白（UP），用自动生化分析仪检测血清胆固醇（CHOL）、白蛋白（ALB）、血尿素氮（BUN）和肌酐（SCr）；用酶联免疫吸附试验试剂盒检测血清白细胞介素-6（IL-6）、IL-1β、肿瘤坏死因子-α（TNF-α）水平；用 HE 染色观察肾组织形态学变化；用实时 RT-PCR 检测肾组织 miR-16 和 MDR1 mRNA 水平；用 Western blot 检测肾组织 NF-κB p65、p-NF-κB p65 和 P-gp170 蛋白表达水平；用双荧光素酶报告基因验证 miR-16 与 NF-κB 的关系。
对照组大鼠肾组织结构正常，无炎性细胞浸润。模型组大鼠肾小球轻度充血，毛细血管狭窄或闭塞，炎性细胞浸润明显。APS 低剂量组和高剂量组大鼠肾小球无明显充血，系膜细胞增生明显减少，炎性细胞减少。与高剂量 APS 组和 APS+miR-16 反义寡核苷酸对照组比较，APS+miR-16 反义寡核苷酸组大鼠肾组织结构损伤更严重。与对照组比较，模型组 UP、CHOL、BUN、SCr、IL-6、IL-1β、TNF-α和 MDR1 mRNA 水平及 p-NF-κB p65 和 P-gp170 蛋白水平均显著升高（均<0.05），ALB 和 miR-16 水平显著降低（均<0.05）。与模型组比较，APS 低剂量组和高剂量组 UP、CHOL、BUN、SCr、IL-6、IL-1β、TNF-α和 MDR1 mRNA 水平及 p-NF-κB p65 和 P-gp170 蛋白水平均显著降低（均<0.05），ALB 和 miR-16 水平均显著升高（均<0.05）。与 APS+miR-16 反义寡核苷酸对照组比较，APS+miR-16 反义寡核苷酸组 UP、CHOL、BUN、SCr、IL-6、IL-1β和 TNF-α水平、MDR1 mRNA 水平及 p-NF-κB p65 和 P-gp170 蛋白水平均显著升高（均<0.05），ALB 和 miR-16 水平均显著降低（均<0.05）。
APS 可通过调控 miR-16/NF-κB 信号通路，影响 MDR1 和 P-gp170 的水平，减少阿霉素肾病大鼠肾组织的炎症反应。
