Duojie Renqing, Chen Min, Li Chenghua, Wang Fu, Wang Xiaobo, Meng Xianli, Cairang Nanjia, Hou Ya
Department of Tibetan Medicine, University of Xizang Medicine.
School of Pharmacy/School of Modern Chinese Medicine Industry, Chengdu University of Traditional Chinese Medicine; TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine.
J Vis Exp. 2025 Jun 27(220). doi: 10.3791/68319.
β-cell failure caused by mitochondrial dysfunction is the core feature of type 2 diabetes mellitus (T2DM). Tibetan medicine Jiangtangle, composed of Berberis kansuensis, Curcumaelongae Rhizoma, Malvae Fructus, Tribuli Fructus, Thlaspi arvense L., and Phyllanthus emblica L., can be used to remedy T2DM. However, its effects on mitochondrial function remain unclear. This study aims to clarify the pharmacological effects of Tibetan medicine Jiangtangle on mitochondria. At 7 days after pre-administration of Tibetan medicine Jiangtangle, rat serum was obtained by abdominal aorta blood collection. Rat pancreatic islet β-cells (RIN-m5F) were divided into control, model, and Jiangtangle-containing serum groups. The model and Jiangtangle-containing serum groups were induced for injury with DMEM medium containing 50 mM glucose for 24 h. The control group was treated with DMEM medium containing no glucose for 24 h. Then, medium was added to the control and model groups, while Jiangtangle-containing serum groups were incubated for 24 h with medium containing 2.5%, 5%, and 10% Jiangtangle-containing serum. The results of UPLC-ESI-Q-Orbitrap-MS indicated 90 compounds in Jiangtangle, among which 6 prototypical compounds entering the blood. CCK 8 was used to measure cell viability, Seahorse XFe was employed to evaluate mitochondrial energy metabolism and the effect of Jiangtangle-containing serum on the RIN-m5F cells hyperglycemic model was evaluated. Jiangtangle-containing serum augmented RIN-m5F cell viability, mitochondrial ATP level, glycolytic ATP level, basal glycolysis, compensatory glycolysis, OCR, proton leak, and maximal respiration.
线粒体功能障碍引起的β细胞功能衰竭是2型糖尿病(T2DM)的核心特征。藏药降糖乐由甘肃小檗、姜黄、冬葵子、刺蒺藜、遏蓝菜和余甘子组成,可用于治疗T2DM。然而,其对线粒体功能的影响尚不清楚。本研究旨在阐明藏药降糖乐对线粒体的药理作用。预先给予藏药降糖乐7天后,通过腹主动脉采血获取大鼠血清。大鼠胰岛β细胞(RIN-m5F)分为对照组、模型组和含降糖乐血清组。模型组和含降糖乐血清组用含50 mM葡萄糖的DMEM培养基诱导损伤24小时。对照组用不含葡萄糖的DMEM培养基处理24小时。然后,向对照组和模型组加入培养基,而含降糖乐血清组分别用含2.5%、5%和10%含降糖乐血清的培养基孵育24小时。超高效液相色谱-电喷雾电离-四极杆-轨道阱质谱(UPLC-ESI-Q-Orbitrap-MS)结果表明,降糖乐中含有90种化合物,其中6种原型化合物进入血液。采用CCK 8检测细胞活力,利用Seahorse XFe评估线粒体能量代谢,并评估含降糖乐血清对RIN-m5F细胞高血糖模型的影响。含降糖乐血清可提高RIN-m5F细胞活力、线粒体ATP水平、糖酵解ATP水平、基础糖酵解、代偿性糖酵解、氧消耗率、质子泄漏和最大呼吸。