College of Basic Medicine, Henan University of Traditional Chinese Medicine, Zhengzhou 450000, China.
The Second School of Clinical Medicine of Henan University of Traditional Chinese Medicine, Zhengzhou 450011, China.
Curr Pharm Biotechnol. 2020;21(12):1204-1212. doi: 10.2174/1389201021666200416094318.
To investigate the effect of Danggui-Shaoyao-San (DSS)-containing serum on the renal tubular Epithelial-Mesenchymal Transition (EMT) of Diabetic Nephropathy (DN) in high glucose- induced HK-2 cells and its mechanism.
20 rats were randomly divided into four groups: blank control group, DSS low dose group (DSS-L), DSS middle dose group (DSS-M), and DSS high dose group (DSS-H). DSS was administrated to the corresponding group (7g/kg/d, 14g/kg/d and 21g/kg/d) for 7 consecutive days, and the same volume of saline was given to the blank control group by gavage. The rat drug-containing serum was successfully prepared. HK-2 cells were divided into five groups: blank control group, model group, DSS-L, DSS-M, DSS-H, according to the corresponding drug and dose of each treatment group. Protein and mRNA levels of Jagged1, Notch1, Hes5, Notch Intracellular Domain (NICD), E-cadherin, alpha- Smooth Muscle Actin (α-SMA) and vimentin at 24h, 48h and 72h were detected by Western Blot and RT-qPCR.
The protein and mRNA levels of Jagged1, Notch1, Hes5, NICD, α-SMA and vimentin in the treatment groups were remarkably decreased compared with the model group (P<0.05), and the protein and mRNA levels of E-cadherin were notably increased (P<0.05) by Western Blot and RT-qPCR.
Our results demonstrated that DSS could prevent DN by ameliorating renal tubular EMT through inhibition of the Notch signaling pathway.
探讨当归芍药散（DSS）含药血清对高糖诱导的 HK-2 细胞肾小管上皮间质转化（EMT）的影响及作用机制。
20 只大鼠随机分为 4 组：空白对照组、DSS 低剂量组（DSS-L）、DSS 中剂量组（DSS-M）、DSS 高剂量组（DSS-H）。连续 7 天，相应组给予 DSS（7g/kg/d、14g/kg/d 和 21g/kg/d）灌胃，空白对照组给予等体积生理盐水。成功制备大鼠含药血清。根据各治疗组的相应药物和剂量，将 HK-2 细胞分为 5 组：空白对照组、模型组、DSS-L、DSS-M、DSS-H。采用 Western blot 和 RT-qPCR 检测 Jagged1、Notch1、Hes5、Notch 细胞内结构域（NICD）、E-钙黏蛋白、α-平滑肌肌动蛋白（α-SMA）和波形蛋白在 24h、48h 和 72h 的蛋白和 mRNA 水平。
与模型组相比，治疗组 Jagged1、Notch1、Hes5、NICD、α-SMA 和波形蛋白的蛋白和 mRNA 水平明显降低（P<0.05），E-钙黏蛋白的蛋白和 mRNA 水平明显升高（P<0.05）。
DSS 可通过抑制 Notch 信号通路改善肾小管 EMT，从而预防糖尿病肾病。
