Department of Nephrology, The People's Hospital of Guangxi Zhuang Autonomous Region, Qingxiu, China.
Department of Sci-Tech Novelty Retrieval, Guangxi Medical Information Institute, Qingxiu, China.
Kidney Blood Press Res. 2019;44(6):1339-1351. doi: 10.1159/000502524. Epub 2019 Oct 29.
This study aimed to investigate the effect of norcantharidin (NCTD) on human mesangial cells (HMCs) apoptosis in vitro and further examine its molecular mechanism.
HMCs were divided into 5 groups: control group, 25% fetal bovine serum (FBS)-treated group, and NCTD groups (NCTD [2.5, 5 and 10 µg/mL] + 25% FBS, respectively). Cell proliferation was determined by MTT assay, while apoptosis was evaluated by Hoechest 33258 staining, the level of cytochrome c, immunohistochemistry, and apoptotic-related proteins/gene expression.
Cell viability was inhibited in NCTD-treated HMCs in a dose-dependent manner. The number of apoptotic cells and the content of cytochrome c were significantly increased by NCTD treatment but that of mitochondrial membrane was decreased. Moreover, the expression of bcl-2 and caspase-3 was prompted by NCTD, but the expression of bax, MMP-2, and MMP-9 in 25% FBS-treated HMCs was inhibited. In addition, NCTD markedly unregulated the expression of apoptosis-related gene/protein, including p-Erk1/2, phosphorylated-Jun N-terminal kinase (JNK), p-p38, and p53.
NCTD enhances 25% FBS-treated HMC apoptosis in vitro, and this effect may be attributed to the modulation of the ERK, JNK, and p38 mitogen-activated protein kinase signaling pathways.
本研究旨在探讨去甲斑蝥素（NCTD）对体外人肾小球系膜细胞（HMCs）凋亡的影响，并进一步探讨其分子机制。
将 HMC 分为 5 组：对照组、25%胎牛血清（FBS）处理组和 NCTD 组（NCTD [2.5、5 和 10 µg/mL] + 25% FBS）。通过 MTT 法测定细胞增殖，Hoechest 33258 染色评估细胞凋亡，检测细胞色素 c 水平、免疫组化和凋亡相关蛋白/基因表达。
NCTD 处理的 HMC 细胞活力呈剂量依赖性抑制。NCTD 处理可显著增加细胞凋亡数和细胞色素 c 含量，减少线粒体膜的含量。此外，NCTD 可促进 bcl-2 和 caspase-3 的表达，但抑制 25% FBS 处理的 HMC 中 bax、MMP-2 和 MMP-9 的表达。此外，NCTD 明显调节凋亡相关基因/蛋白的表达，包括 p-Erk1/2、磷酸化-Jun N 末端激酶（JNK）、p-p38 和 p53。
NCTD 增强了体外 25% FBS 处理的 HMC 凋亡，这种作用可能归因于 ERK、JNK 和 p38 丝裂原激活蛋白激酶信号通路的调节。
