College of Pharmacy, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China (mainland).
Department of Traditional Chinese Medicine (TCM) Pharmacy, Zhejiang Integrated Traditional and Western Medicine Hospital, Hangzhou, Zhejiang, China (mainland).
Med Sci Monit. 2020 Jun 3;26:e922943. doi: 10.12659/MSM.922943.
BACKGROUND Shen Qi Wan (SQW) as a well-known formula for the amelioration of kidney yang deficiency syndrome (KYDS), and it has been widely employed in traditional Chinese medicine (TCM). This study aimed to investigate the effect and underlying mechanism of SQW medicated serum on proliferation and migration in NRK-52E cells. MATERIAL AND METHODS We employed the real-time cell analysis (RTCA) system to investigate the effect of SQW medicated serum on proliferation and migration in NRK-52E cells. In addition, the migration was further investigated by using a wound-healing assay. The mRNA and protein expression level of aquaporin 1 (AQP1) of NRK-52E cells with SQW medicated serum-treated were quantified by real-time quantitative polymerase chain reaction (q-PCR) and western blot assay, respectively. Furthermore, NRK-52E cells were transfected with lentivirus AQP1-RNAi to assess migratory cell abilities in vitro. RESULTS The migratory abilities of NRK-52E cells were significantly increased after SQW medicated serum treatment (P<0.05), and no significant difference in cell proliferation. In addition, SQW medicated serum was significantly upregulated the mRNA and protein expression level of AQP1 in NRK-52E cells (P<0.05). Additionally, the in vitro metastasis test proved that knockdown of AQP1 suppressed migratory abilities according to RTCA and wound healing test while was reversed by SQW medicated serum (P<0.05). CONCLUSIONS Our study demonstrates that SQW medicated serum effectively promotes the migration of NRK-52E cells by increasing AQP1 expression, and AQP1 may be as a therapeutic target of SQW for renal injury treatment under KYDS.
参芪丸(SQW)是一种治疗肾阳虚证(KYDS)的著名方剂,已在中医中广泛应用。本研究旨在探讨 SQW 含药血清对 NRK-52E 细胞增殖和迁移的作用及机制。
我们采用实时细胞分析(RTCA)系统研究 SQW 含药血清对 NRK-52E 细胞增殖和迁移的影响。此外,还通过划痕愈合试验进一步研究了迁移作用。采用实时定量聚合酶链反应(q-PCR)和 Western blot 法分别定量检测 SQW 含药血清处理的 NRK-52E 细胞中水通道蛋白 1(AQP1)的 mRNA 和蛋白表达水平。此外,还通过慢病毒 AQP1-RNAi 转染 NRK-52E 细胞,评估体外迁移细胞能力。
SQW 含药血清处理后 NRK-52E 细胞的迁移能力显著增加(P<0.05),细胞增殖无显著差异。此外,SQW 含药血清明显上调 NRK-52E 细胞 AQP1 的 mRNA 和蛋白表达水平(P<0.05)。此外,体外转移试验证明,根据 RTCA 和划痕愈合试验,AQP1 的敲低抑制了迁移能力,而 SQW 含药血清则逆转了这一现象(P<0.05)。
我们的研究表明,SQW 含药血清通过增加 AQP1 的表达有效促进 NRK-52E 细胞的迁移,AQP1 可能是 SQW 在 KYDS 下治疗肾损伤的治疗靶点。