Gao Jingying, Xia Lixia, Wei Yuanyuan
Department of Pediatrics, Shanxi Medical University, Taiyuan 030001, China.
Pediatric Internal Medicine, Children's Hospital of Shanxi Province, Shanxi Medical University, Taiyuan 030001, China.
Korean J Physiol Pharmacol. 2022 May 1;26(3):165-174. doi: 10.4196/kjpp.2022.26.3.165.
As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.
由于氧化苦参碱调节葡萄糖代谢的机制尚不清楚，本研究探讨了氧化苦参碱对INS-1细胞焦亡的影响。采用流式细胞术检测细胞焦亡和活性氧（ROS）生成。还通过透射电子显微镜和乳酸脱氢酶（LDH）释放来研究细胞焦亡。使用蛋白质印迹法检测蛋白质水平，并通过酶联免疫吸附测定法检测白细胞介素（IL）-1β和IL-18的分泌。还评估了半胱天冬酶-1活性以及核因子κB（NF-κB）和核因子（红系衍生2）样2蛋白（Nrf2）的DNA结合活性。在高糖高脂处理的INS-1细胞（HG + PA）中，半胱天冬酶-1活性和LDH含量以及含Nod样受体家族吡咯结构域3、Gsdmd-N、半胱天冬酶-1、含CARD的凋亡相关斑点样蛋白、IL-1β和IL-18水平均升高。此外，P65蛋白水平在细胞核中升高但在细胞质中降低。氧化苦参碱减弱了这些作用并抑制了高糖高脂诱导的ROS生成。氧化苦参碱处理后，HG + PA细胞中核Nrf2和血红素加氧酶-1（HO-1）水平的升高进一步增加，而细胞质中Nrf2和Keleh样ECH相关蛋白水平降低。此外，氧化苦参碱降低了HG + PA细胞中p65升高的转录活性，而Nrf2的转录活性增加。结果表明，氧化苦参碱抑制INS-1细胞焦亡是其调节葡萄糖代谢的关键因素，涉及抑制NF-κB途径和激活Nrf2/HO-1途径。
