Department of Endocrinology, Guangzhou University of Chinese Medicine, No.1 South Second Street, Fei'e West Road, Baiyun District, Guangzhou, 510405, Guangdong Province, China.
Mol Biotechnol. 2023 Oct;65(10):1632-1643. doi: 10.1007/s12033-022-00651-5. Epub 2023 Feb 3.
Diabetic encephalopathy (DE) is one of the major chronic complications of diabetes mellitus. This study aims to investigate the inhibitory effect of berberine (BBR) on the damage of PC12 cells induced by high glucose (HG). Differentiated PC12 cells were treated with different concentrations of glucose/BBR. The cell morphology, cell viability, lactate dehydrogenase (LDH) activity, apoptosis, oxidative stress (OS), mitochondrial structure, mitochondrial membrane potential (MMP), mitochondrial complex I-V activity, and adenosine triphosphate (ATP) levels were evaluated. The mRNA and protein levels of the Keap1/Nrf2/ARE pathway-related genes were assessed by RT-qPCR and Western blot. High-dose BBR and HG jointly treated-PC12 cells were treated with Nrf2-specific inhibitor ML385 to further verify whether Nrf2 was the target of BBR. The results showed that BBR inhibited cell damage, OS, and mitochondrial dysfunction induced by HG. The inhibitory effect of high BBR was more significant. The Keap1/Nrf2/ARE pathway was inhibited in PC12 cells induced by HG. BBR could activate the Keap1/Nrf2/ARE pathway, thus up-regulating the expression levels of antioxidant enzymes. ML385 antagonized the ameliorating effect of BBR on OS and mitochondrial dysfunction. The conclusion is that BBR can activate the Keap1/Nrf2/ARE pathway, upregulate the expression patterns of antioxidant enzymes, and reduce cell damage, OS, and mitochondrial dysfunction of PC12 cells induced by HG.
糖尿病性脑病（DE）是糖尿病的主要慢性并发症之一。本研究旨在探讨小檗碱（BBR）对高糖（HG）诱导的 PC12 细胞损伤的抑制作用。用不同浓度的葡萄糖/BBR 处理分化的 PC12 细胞。评估细胞形态、细胞活力、乳酸脱氢酶（LDH）活性、细胞凋亡、氧化应激（OS）、线粒体结构、线粒体膜电位（MMP）、线粒体复合物 I-V 活性和三磷酸腺苷（ATP）水平。通过 RT-qPCR 和 Western blot 评估 Keap1/Nrf2/ARE 通路相关基因的 mRNA 和蛋白水平。用 Nrf2 特异性抑制剂 ML385 处理高剂量 BBR 和 HG 共同处理的 PC12 细胞，进一步验证 Nrf2 是否是 BBR 的靶点。结果表明，BBR 抑制 HG 诱导的细胞损伤、OS 和线粒体功能障碍。高浓度 BBR 的抑制作用更为显著。HG 诱导的 PC12 细胞中 Keap1/Nrf2/ARE 通路受到抑制。BBR 可激活 Keap1/Nrf2/ARE 通路，从而上调抗氧化酶的表达水平。ML385 拮抗了 BBR 对 OS 和线粒体功能障碍的改善作用。结论是，BBR 可激活 Keap1/Nrf2/ARE 通路，上调抗氧化酶的表达模式，减轻 HG 诱导的 PC12 细胞的细胞损伤、OS 和线粒体功能障碍。
