Chen Hongshu, Zhou Xiaojie, Yang Yuanxiao, Feng Yaorong
The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang 310006, China.
Academy of Chinese Medical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, China.
Evid Based Complement Alternat Med. 2023 Feb 6;2023:3352353. doi: 10.1155/2023/3352353. eCollection 2023.
Shen Qi Wan (SQW) is the most classic prescription for the clinical therapy of chronic kidney disease in China. Nevertheless, the function of SQW in renal interstitial fibrosis (RIF) has not been clearly clarified. Our purpose was to explore the protective function of SQW on RIF.
After intervention with SQW-containing serum alone at increasing concentrations (2.5, 5, and 10%) or in combination with siNotch1, the transforming growth factor-beta (TGF-)-induced HK-2 cell viability, extracellular matrix (ECM)-, epithelial-mesenchymal transition (EMT), and Notch1 pathway-associated protein expressions were assessed by cell counting kit-8, qRT-PCR, western blot, and immunofluorescence assays.
SQW-containing serum intensified the viability of TGF--mediated HK-2 cells. Besides, it augmented the collagen II and E-cadherin levels, and weakened the fibronectin, -SMA, vimentin, N-cadherin, and collagen I levels in HK-2 cells triggered by TGF-. Moreover, it is found that TGF- led to the upregulation of Notch1, Jag1, HEY1, HES1, and TGF- in HK-2 cells, which was partially offset by SQW-containing serum. Furthermore, cotreatment of SQW-containing serum and Notch1 knockdown further apparently alleviated the Notch1, vimentin, N-cadherin, collagen I, and fibronectin levels in HK-2 cells induced by TGF-.
Collectively, these findings elucidated that SQW-containing serum attenuated RIF via restraining EMT through the repression of the Notch1 pathway.
肾芪丸(SQW)是中国慢性肾脏病临床治疗中最经典的方剂。然而,SQW在肾间质纤维化(RIF)中的作用尚未明确阐明。我们的目的是探讨SQW对RIF的保护作用。
用不同浓度(2.5%、5%和10%)的含SQW血清单独干预,或与siNotch1联合干预后,通过细胞计数试剂盒-8、qRT-PCR、蛋白质免疫印迹和免疫荧光分析评估转化生长因子-β(TGF-β)诱导的HK-2细胞活力、细胞外基质(ECM)、上皮-间质转化(EMT)及Notch1信号通路相关蛋白表达。
含SQW血清增强了TGF-β介导的HK-2细胞活力。此外,它提高了HK-2细胞中Ⅱ型胶原蛋白和E-钙黏蛋白水平,并降低了TGF-β诱导的HK-2细胞中纤连蛋白、α-平滑肌肌动蛋白(α-SMA)、波形蛋白、N-钙黏蛋白和Ⅰ型胶原蛋白水平。此外,发现TGF-β导致HK-2细胞中Notch1、Jag1、HEY1、HES1和TGF-β上调,而含SQW血清可部分抵消这种上调。此外,含SQW血清与Notch1基因敲低共同处理进一步明显降低了TGF-β诱导的HK-2细胞中Notch1、波形蛋白、N-钙黏蛋白、Ⅰ型胶原蛋白和纤连蛋白水平。
总体而言,这些发现表明含SQW血清通过抑制Notch1信号通路抑制EMT,从而减轻RIF。