| 引用本文:李祥宇1，崔涛1，姜月华2，李伟2.黄芪-丹参药对通过下调miR-466b-5p改善高血压肾损害[J].世界中医药,2023,(05):. |
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| 黄芪-丹参药对通过下调miR-466b-5p改善高血压肾损害 |
| Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma Herbal Pair Reduces Hypertensive Renal Damage by Down-regulating MiR-466b-5p |
| 投稿时间：2022-02-07 |
| DOI：10.3969/j.issn.1673-7202.2023.05.004 |
| 中文关键词: 高血压肾损害 小鼠 黄芪-丹参药对 微RNA测序 miR-466b-5p 肾脏病理 透明质酸酶2 |
| English Keywords:Hypertensive renal damage Mice Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma herbal pair MiRNA sequencing MiR-466b-5p Renal pathology HAS2 |
| 基金项目:国家自然科学基金面上项目（81673812） |
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| 中文摘要: |
| 目的：探讨黄芪-丹参药对通过调节微小核糖核酸-466b-5P（miR-466b-5p）改善高血压肾损害的机制。方法：基于微RNA（miRNA）测序寻找自发性高血压大鼠与Wistar-京都鼠（WKY）的差异基因，构建腺相关病毒转染小鼠，24只C57BL/6小鼠，随机选取12只尾静脉注射小鼠腺相关病毒miR-466b-5P（rAAV-miR-466b-5P）构建miR-466b-5p过表达组，余12只作为空白对照组注射腺相关病毒-9（AAV-9）空载体，转染6周后再各随机分为2组，每组6只，A组（黄芪-丹参+miR-466b-5P过表达组）和C组（黄芪-丹参+空白对照组）以黄芪配方颗粒：2.036 g/kg+丹参配方颗粒：0.255 g/kg灌胃；B组（miR-466b-5P过表达组）和D组（空白对照组）以相同体积生理盐水灌胃；灌胃28 d后观察各组小鼠尿β2-微球蛋白（β2-MG）、N-乙酰-β-D-葡萄糖苷酶（NAG）、微量白蛋白（mALB），血清胱抑素C（Cys-C）、血管紧张素Ⅱ（AngⅡ）、C反应蛋白（CRP）水平和肾脏病理，进行实时荧光定量(RT-qPCR)与蛋白质印迹法(Western Blotting)寻找其靶基因。结果：1）基于前期miRNA测序选定了miR-466b-5p；2）与D组比较，B组小鼠尿β2-MG、NAG、mALB、血清Cys-C、AngⅡ、CRP均显著升高（P<0.01）；与B组比较，A组小鼠血清Cys-C、CRP明显升高（P<0.05），尿β2-MG、NAG、mALB、血清AngⅡ无明显改变（P>0.05）；3）与D组比较，B组小鼠出现明显肾小球硬化、小管纤维化，A组较B组有明显改善；4）与D组比较，B组小鼠透明质酸酶2（HAS2）明显下调，而经黄芪-丹参干预后有所回调。结论：miR-466b-5p的过表达会导致高血压肾损害的发生，黄芪-丹参药对通过靶向HAS2下调miR-466b-5p改善高血压肾损害。 |
| English Summary: |
| To explore the mechanism of Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma herbal pair in reducing hypertensive renal damage by regulating miR-466b-5p.Methods:The differential genes between spontaneously hypertensive rats(SHR) and Wistar-Kyoto(WKY) rats were found by miRNA sequencing to construct adeno-associated virus-transfected rats.Twelve out of 24 C57BL/6 mice were randomly selected to be injected with adeno-associated virus miR-466b-5P(rAAV-miR-466b-5P) into the tail vein to construct a miR-466b-5p overexpression group，and the other 12 mice were assigned to the blank control group and were injected with AAV-9 empty carrier.Six weeks after the transfection，the two groups were randomly subdivided into two groups，with six mice in each group.Group A(Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma+miR-466b-5P overexpression group) and group C(Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma+blank control group) received Huangqi Formula Granules(2.036 g/kg) and Danshen Formula Granules(0.255 g/kg) by gavage.Group B(miR-466b-5P overexpression group) and group D(blank control group) were received the same volume of saline by gavage.After 28 d of intragastric administration，the urine β2-microglobulin(β2-MG)，N-acetyl-β-D-glucosidase(NAG)，microalbumin(mALB)，serum cystatin C(Cys-C)，angiotensin Ⅱ(AngⅡ)，and C-reactive protein(CRP) levels and renal pathology of mice were observed in each group，and Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and Western Blotting assay were performed to find the target genes.Results:1) MiR-466b-5p was selected based on previous miRNA sequencing. 2) The urine β2-MG，NAG，mALB，serum Cys-C，AngⅡ，and CRP levels in group B were higher than those in the group D(P<0.01).Compared with the results in group B，the serum Cys-C and CRP levels increased in group A(P<0.01)，while the urine β2-MG，NAG，mALB，and serum AngⅡ levels did not change significantly(P>0.05). 3) Compared with group D，group B showed significant glomerulosclerosis and tubular fibrosis，and group A showed significant improvement compared with group B. 4) The hyaluronidase 2(HAS2) was significantly down-regulated in group B，which was reversed after Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma intervention as compared with that in the group D.Conclusion:The overexpression of miR-466b-5p can cause hypertensive renal damage，and Astragali Radix-Salviae Miltiorrhizae Radix et Rhizoma can down-regulate miR-466b-5p by targeting HAS2 to reduce hypertensive renal damage. |
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