Yan Zijie, Zhang Lin, Kang Yu, Liu Shuman, Li Xiaoyan, Li Lidan, Rui Kai, Xiao Man, Xie Yiqiang
College of Traditional Chinese Medicine, Hainan Medical University, Haikou, China.
Heilongjiang Academy of Traditional Chinese Medicine, Harbin, China.
Front Med (Lausanne). 2024 Jul 3;11:1424644. doi: 10.3389/fmed.2024.1424644. eCollection 2024.
Diabetic kidney disease (DKD), one of the microvascular complications in patients with diabetes mellitus, is a common cause of end-stage renal disease. Cellular senescence is believed to be an essential participant in the pathogenesis of DKD. Although there is evidence that Alpiniae oxyphyllae fructus (AOF) can ameliorate DKD progression and organismal senescence, its ability to ameliorate renal cellular senescence in DKD as well as active components and molecular mechanisms remain to be explored.
This study aimed to investigate the role of AOF in the treatment of cellular senescence in DKD and to explore its active components and potential molecular mechanisms.
The pharmacological efficacy of AOF in ameliorating cellular senescence in DKD was assessed by establishing DKD mouse models and HK-2 cells under high glucose stress. UHPLC-QTOF-MS was used to screen the active compounds in AOF, which were used in conjunction with network pharmacology to predict the molecular mechanism of AOF in the treatment of cellular senescence in DKD.
experiments showed that AOF reduced GLU, mAlb, Scr, BUN, MDA, SOD levels, and ameliorated renal pathological damage and renal cell senescence in DKD mice. experiments showed that AOF-containing serum improved the decline in HK-2 cell viability and alleviated cellular senescence under high glucose intervention. The results of the UHPLC-QTOF-MS screened 26 active compounds of AOF. The network pharmacological analyses revealed that Cubebin, 2',6'-dihydroxy-4'-methoxydihydrochalcone, Chalcone base + 3O,1Prenyl, Batatasin IV, and Lucidenolactone were the five core compounds and TP53, SRC, STAT3, PIK3CA, and AKT1 are the five core targets of AOF in the treatment of DKD. Molecular docking simulation results showed that the five core compounds had good binding ability to the five core targets. Western blot validated the network pharmacological prediction results and showed that AOF and AOF-containing serum down-regulate the expression of TP53, and phosphorylation of SRC, STAT3, PIK3CA, and AKT.
Our study shows that AOF may delay the development of cellular senescence in DKD by down-regulating the levels of TP53, and phosphorylation of SRC, STAT3, PIK3CA, and AKT.
糖尿病肾病（DKD）是糖尿病患者微血管并发症之一，是终末期肾病的常见病因。细胞衰老被认为是DKD发病机制中的重要参与者。尽管有证据表明益智仁（AOF）可改善DKD进展和机体衰老，但其改善DKD肾细胞衰老的能力以及活性成分和分子机制仍有待探索。
本研究旨在探讨AOF在治疗DKD细胞衰老中的作用，并探索其活性成分和潜在分子机制。
通过建立DKD小鼠模型和高糖应激下的HK-2细胞，评估AOF改善DKD细胞衰老的药理作用。采用超高效液相色谱-四极杆飞行时间质谱（UHPLC-QTOF-MS）筛选AOF中的活性化合物，并结合网络药理学预测AOF治疗DKD细胞衰老的分子机制。
实验表明，AOF降低了DKD小鼠的血糖（GLU）、微量白蛋白（mAlb）、血肌酐（Scr）、尿素氮（BUN）、丙二醛（MDA）水平，提高了超氧化物歧化酶（SOD）水平，改善了肾脏病理损伤和肾细胞衰老。实验表明，含AOF血清改善了高糖干预下HK-2细胞活力的下降，并减轻了细胞衰老。UHPLC-QTOF-MS结果筛选出AOF的26种活性化合物。网络药理学分析显示，二氢山姜素、2',6'-二羟基-4'-甲氧基二氢查耳酮、查耳酮碱+3O,1异戊烯基、落葵薯素IV和亮菌内酯是五种核心化合物，而TP53、SRC、信号转导和转录激活因子3（STAT3）、磷脂酰肌醇-3激酶催化亚基α（PIK3CA）和蛋白激酶B（AKT1）是AOF治疗DKD的五个核心靶点。分子对接模拟结果表明，这五种核心化合物与五个核心靶点具有良好的结合能力。蛋白质免疫印迹法验证了网络药理学预测结果，表明AOF和含AOF血清下调了TP53的表达以及SRC、STAT3、PIK3CA和AKT的磷酸化水平。
我们的研究表明，AOF可能通过下调TP53水平以及SRC、STAT3、PIK3CA和AKT的磷酸化来延缓DKD细胞衰老的发展。
