| 孙佳,沈军.抗microRNA-21治疗对糖尿病肾病小鼠模型肾脏疾病进展的实验研究[J].浙江中西医结合杂志,2025,35(1): |
| 抗microRNA-21治疗对糖尿病肾病小鼠模型肾脏疾病进展的实验研究 |
| Experimental study of anti-microRNA-21 therapy on renal disease progression in diabetic nephropathy mice model |
| 投稿时间：2024-01-29 修订日期：2024-11-14 |
| DOI： |
| 中文关键词: 糖尿病肾病 小鼠 肾脏损伤 microRNA-21 细胞凋亡 |
| 英文关键词:Diabetic nephropathy Mice Kidney damage microRNA-21 Apoptosis. |
| 基金项目:余杭区医疗卫生科研项目(2019伦审研024号) |
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| 中文摘要: |
| 目的 探讨抗microRNA-21(miRNA-21)治疗对糖尿病肾病(diabeticnephropathy,DN)小鼠模型肾脏损伤的作用。方法 通过腹腔注射链脲佐菌素构建DN模型小鼠。小鼠分为正常组、DN 组、DN+miR-21-MM-ASO(antisense oligonucleotides)组和DN+miR-21-ASO组。全自动生化分析仪检测小鼠24 h尿蛋白(Urine protein,Upro)、血肌酐(Serum creatinine,Scr)和血清尿素氮(blood urea nitrogen,BUN)的含量。PAS染色观察小鼠肾脏糖原的沉积,苏木素-伊红染色观察小鼠肾脏的病理学变化和TUNEL染色观察小鼠肾脏组织的细胞凋亡情况。实时定量PCR检测小鼠肾脏组织中miRNA-21的相对表达水平。Western blot检测Bcl-2、Bax和cleaved caspase-3蛋白的表达水平。结果 与正常组相比,DN组小鼠肾脏组织中miRNA-21表达显著上调(P < 0.05)且24 h Upro含量、Scr和BUN水平显著增高；然而,DN组小鼠腹腔注射LNA(locked nucleic acid)-miRNA-21 ASO后,与DN+miR-21-MM-ASO组相比,DN+miR-21-ASO组小鼠肾脏组织中miRNA-21的表达受到抑制(P < 0.05)并且小鼠的24 h Upro、Scr和BUN水平明显降低。PAS 染色显示,与DN+miR-21-MM-ASO组相比,DN+miR-21-ASO组小鼠系膜和基底膜区PAS染色阳性物质明显减少且红染区的区域较少。同时,HE染色表明,与正常组相比,DN组小鼠表现出肾小球体积增大和肾小管刷状缘脱离等肾脏损伤,注射LNA-miRNA-21 ASO后,DN+miR-21-ASO组小鼠肾脏损伤显著改善。此外,TUNEL染色表明,与正常组相比,DN组小鼠肾脏组织细胞凋亡显著增加,而与DN+miR-21-MM-ASO组相比,DN+miR-21-ASO组细胞凋亡显著降低且Western blot检测也显示Bax和cleaved caspase-3表达下调,Bcl-2表达上调。结论 抗microRNA-21治疗可以明显改善糖尿病肾病模型小鼠的肾脏损伤。 |
| 英文摘要: |
| Objective To investigate the effect of anti-microRNA-21 (miRNA-21) treatment on renal injury in diabetic nephropathy (DN) mouse model. Methods DN model mice were constructed by intraperitoneal injection of streptozotocin (STZ). Mice were divided into normal group, DN group, DN+miR-21-MM-ASO (antisense oligonucleotides) group and DN+miR-21-ASO group. The 24 h urinary protein (Upro), serum creatinine (Scr)and blood urea nitrogen(BUN) in mice were detected by automatic biochemical analyzer. PAS staining was used to observe glycogen deposition in mouse kidney, hematoxylin-eosin staining was used to observe pathological changes in mouse kidney, and TUNEL staining was used to observe apoptosis in mouse kidney. The relative expression level of miRNA-21 in mouse kidney tissue was detected by real-time quantitative PCR. The protein expressions of Bcl-2, Bax and cleaved caspase-3 were detected by Western blot. Results Compared with the normal group, the expression of miRNA-21 in renal tissue of DN group was significantly up-regulated (P < 0.05), and the 24 h Upro content, Scr and BUN levels were significantly increased; however, after intraperitoneal injection of LNA (locked nucleic acid) -miRNA-21 ASO in DN group, compared with DN+miR-21-MM-ASO group, the expression of miRNA-21 in kidney tissue of DN+miR-21-ASO group was inhibited (P < 0.05), and the 24 h Upro, Scr and BUN levels of mice were significantly decreased. PAS staining showed that compared with DN+miR-21-MM-ASO group, the PAS positive substances in mesangial and basement membrane areas of DN+miR-21-ASO group were significantly reduced, and the red staining area was less. At the same time, HE staining showed that compared with the normal group, the DN group showed renal injury such as glomerular volume enlargement and brush border detachment of renal tubules. After injection of LNA-miRNA-21 ASO, the renal injury of DN+miR-21-ASO group was significantly improved. In addition, TUNEL staining showed that compared with the normal group, the apoptosis of kidney tissue cells in DN group was significantly increased. Compared with DN+miR-21-MM-ASO group, the apoptosis of kidney tissue cells in DN+miR-21-ASO group was significantly decreased. Western blot detection also showed that Bax and cleaved caspase-3 expressions were down-regulated, and Bcl-2 expression was up-regulated. Conclusion Anti-microRNA-21 treatment can significantly improve renal damage in diabetic nephropathy model mice. |
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