Wang Weisong, Long Hongping, Huang Wei, Zhang Ting, Xie Lihua, Chen Cheng, Liu Jianhe, Xiong Dan, Hu Wei
Graduate School, Hunan University of Chinese Medicine, Changsha, China.
Experiment Center of Medical Innovation, The First Hospital of Hunan University of Chinese Medicine, Changsha, China.
Front Pharmacol. 2020 Dec 3;11:587663. doi: 10.3389/fphar.2020.587663. eCollection 2020.
Diabetic nephropathy (DN), a leading cause of end-stage renal disease, is associated with high morbidity and mortality rates worldwide and the development of new drugs to treat DN is urgently required. Bu-Shen-Huo-Xue (BSHX) decoction is a traditional Chinese herbal formula, made according to traditional Chinese medicine (TCM) theory, and has been used clinically to treat DN. In the present study, we established a high-fat diet/streptozotocin-induced diabetic mouse model and treated the mice with BSHX decoction to verify its therapeutic effects . Ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was applied to analyze the chemical composition and active compounds of BSHX decoction. Markers of podocyte epithelial-mesenchymal transition and the Rac1/PAK1/p38MAPK signaling pathway were evaluated to investigate the mechanism underlying function of BSHX decoction. BSHX decoction effectively alleviated diabetic symptoms, according to analysis of the renal function indicators, serum creatinine, blood urea nitrogen, serum uric acid, and urinary albumin excretion rate, as well as renal histopathology and ultrastructural pathology of DN mice. We identified 67 compounds, including 20 likely active compounds, in BSHX decoction. The podocyte markers, nephrin and podocin, were down-regulated, while the mesenchymal markers, α-SMA and FSP-1, were up-regulated in DN mouse kidney; however, the changes in these markers were reversed on treatment with BSHX decoction. GTP-Rac1 was markedly overexpressed in DN mice and its levels were significantly decreased in response to BSHX decoction. Similarly, levels of p-PAK1 and p-p38MAPK which indicate Rac1 activation, were reduced on treatment with BSHX decoction. Together, our data demonstrated that BSHX decoction ameliorated renal function and podocyte epithelial-mesenchymal transition via inhibiting Rac1/PAK1/p38MAPK signaling pathway in high-fat diet/streptozotocin-induced diabetic mice. Further, we generated a quality control standard and numerous potential active compounds from BSHX decoction for DN.
糖尿病肾病（DN）是终末期肾病的主要原因，在全球范围内发病率和死亡率都很高，因此迫切需要开发治疗DN的新药。补肾活血（BSHX）汤是一种按照中医理论制成的传统中药配方，已在临床上用于治疗DN。在本研究中，我们建立了高脂饮食/链脲佐菌素诱导的糖尿病小鼠模型，并用BSHX汤治疗小鼠以验证其治疗效果。采用超高效液相色谱-四极杆飞行时间质谱联用技术（UPLC-Q-TOF-MS）分析BSHX汤的化学成分和活性成分。评估足细胞上皮-间质转化标志物和Rac1/PAK1/p38MAPK信号通路，以研究BSHX汤发挥作用的机制。根据对肾功能指标、血清肌酐、血尿素氮、血清尿酸和尿白蛋白排泄率的分析，以及DN小鼠的肾脏组织病理学和超微结构病理学，BSHX汤有效缓解了糖尿病症状。我们在BSHX汤中鉴定出67种化合物，其中包括20种可能的活性化合物。在DN小鼠肾脏中，足细胞标志物nephrin和podocin下调，而间质标志物α-SMA和FSP-1上调；然而，用BSHX汤治疗后这些标志物的变化得到逆转。GTP-Rac1在DN小鼠中明显过表达，用BSHX汤处理后其水平显著降低。同样，表明Rac1激活的p-PAK1和p-p38MAPK水平在用BSHX汤处理后降低。总之，我们的数据表明，在高脂饮食/链脲佐菌素诱导的糖尿病小鼠中，BSHX汤通过抑制Rac1/PAK1/p38MAPK信号通路改善肾功能和足细胞上皮-间质转化。此外，我们还为DN生成了BSHX汤的质量控制标准和许多潜在的活性化合物。
