Toxicology and Pharmacology Laboratory, Department of Biotechnology, Faculty of Science and Humanities, SRM Institute of Science and Technology, Chengalpattu District, Kattankulathur 603203, Tamil Nadu, India.
Department of Chemistry, Faculty of Engineering and Technology, SRM Institute of Science and Technology, Chengalpattu District, Kattankulathur 603203, Tamil Nadu, India.
Tissue Cell. 2023 Jun;82:102104. doi: 10.1016/j.tice.2023.102104. Epub 2023 May 6.
Diabetes Mellitus is a metabolic disorder characterized by insulin dysfunction or failure of the pancreatic β-cells to produce insulin resulting in hyperglycemia. Adverse effects of hyperglycemic conditions continue to be common, reducing treatment adherence. Intensified therapies are required for the constant loss of endogenous islet reserve.
This study aimed to evaluate the effect of Nimbin semi-natural analogs (N2, N5, N7, and N8) from A. indica on high glucose-induced ROS and apoptosis with insulin resistance in L6 myotubes evaluated along with Wortmannin and Genistein inhibitors and the expression of key genes in the insulin signalling pathway.
The analogs were screened for anti-oxidant and anti-diabetic activity using cell-free assays; The ability of analogs to suppress ROS and prevent apoptosis induced by High glucose and uptake glucose and glycogen storage in L6 myotubes was evaluated using DCFH-DA, AO-PI and 2NBDG staining. Further, the glucose uptake was performed in the presence of Insulin Receptor Tyrosine Kinase (IRTK) inhibitors, and the expression of key genes PI3K, Glut-4, GS and IRTK in the insulin signalling pathway were evaluated.
The Nimbin analogs were not toxic to the L6 cells, and the analogs could scavenge ROS and suppress cellular damage induced due to high glucose. Enhanced glucose uptake was observed in N2, N5 and N7 compared to N8. The maximum activity of optimum concentration was found to be 100 μM. The N2, N5 and N7 showed an increase in IRTK, which is equivalent to insulin at a concentration of 100 µM. The IRTK inhibitor with Genistein (50 µM) confirmed the presence of IRTK-dependent glucose transport activation; it also supports the expression of key genes PI3K, Glut-4, GS and IRTK. As a result of PI3K activation, N2, N5, and N7 exhibited the insulin-mimetic effect by enhancing glucose uptake and glycogen conversion regulating glucose metabolism.
N2, N5 and N7 could therapeutically benefit against insulin resistance by glucose metabolism modulation, insulin secretion, β-cell stimulation, inhibition of gluconeogenic enzymes and ROS protection.
糖尿病是一种代谢紊乱，其特征是胰岛素功能障碍或胰腺β细胞无法产生胰岛素，导致高血糖。高血糖状态的不良后果仍然很常见，降低了治疗的依从性。需要强化治疗以弥补内源性胰岛储备的不断损失。
本研究旨在评估从印楝中提取的 Nimbin 半天然类似物（N2、N5、N7 和 N8）对高葡萄糖诱导的 ROS 和 L6 肌管胰岛素抵抗相关的细胞凋亡的影响，同时评估 Wortmannin 和 Genistein 抑制剂以及胰岛素信号通路关键基因的表达。
使用无细胞测定法筛选类似物的抗氧化和抗糖尿病活性；通过 DCFH-DA、AO-PI 和 2NBDG 染色评估类似物抑制高葡萄糖诱导的 ROS 和预防细胞凋亡以及在 L6 肌管中摄取葡萄糖和糖原储存的能力。此外，在胰岛素受体酪氨酸激酶（IRTK）抑制剂存在的情况下进行葡萄糖摄取，评估胰岛素信号通路中关键基因 PI3K、Glut-4、GS 和 IRTK 的表达。
Nimbin 类似物对 L6 细胞没有毒性，类似物可以清除 ROS 并抑制高葡萄糖引起的细胞损伤。与 N8 相比，N2、N5 和 N7 观察到葡萄糖摄取增加。发现最佳浓度的最大活性为 100 μM。N2、N5 和 N7 显示 IRTK 增加，相当于 100 μM 胰岛素的作用。用 Genistein（50 μM）作为 IRTK 抑制剂证实了 IRTK 依赖性葡萄糖转运激活的存在；它还支持关键基因 PI3K、Glut-4、GS 和 IRTK 的表达。由于 PI3K 激活，N2、N5 和 N7 通过增强葡萄糖摄取和调节葡萄糖代谢的糖原转化表现出胰岛素样作用。
N2、N5 和 N7 可通过调节葡萄糖代谢、胰岛素分泌、β细胞刺激、抑制糖异生酶和保护 ROS 来治疗胰岛素抵抗，从而带来获益。
