Lai Xihu, Kang Xincong, Zeng Luman, Li Jian, Yang Yan, Liu Dongbo
Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Agricultural University, Changsha, Hunan 410128, P, R, China.
BMC Complement Altern Med. 2014 Jan 9;14:10. doi: 10.1186/1472-6882-14-10.
Excessive apoptosis of β-cell is closely related to diabetes mellitus. Chronic exposure to high glucose causes β-cell dysfunction and apoptosis in diabetes. Thorn grape (Vitis davidii Foex.) has been used to treat diabetes in Traditional Chinese medicine for many years. In our previous research, thorn grape seeds oil (TGSO) showed promising anti-diabetic effects in animal models. However, it is unknown whether TGSO played an anti-apoptotic role in the anti-diabetic effects and the mechanism regarding signal transduction pathway is unclear either.
The rattus pancreatic β-cell line RIN-m5F was treated with/without TGSO which was extracted by supercritical carbon dioxide (CO2) fluid extraction and analyzed by Gas Chromatography/Mass Spectrometry (GC/MS). Cell apoptosis was detected by fluorescence activated cell sorting (FACS), insulin secretion was assayed by Enzyme-Linked Immunosorbent Assay (ELISA), and the apoptosis-related genes expressions were evaluated by quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR).
TGSO, containing 87.02% unsaturated fatty acids (UFAs), significantly reduced pancreatic β-cell apoptosis and protected the insulin secretion impaired by high glucose. The expressions of pro-apoptotic genes such as iNOS, Caspase-3, ATF-3, JNK, p38 and Fas were down-regulated while the anti-apoptotic genes Akt and Bcl-2/Bax were up-regulated.
The results indicated that TGSO protected β-cells from high glucose-induced apoptosis and its protective activity may be linked to mitochondrial pathway, endoplasmic reticulum (ER) stress pathway and Fas signal pathway, which implied that TGSO might be an effective complementary or alternative medicine to reduce β-cell apoptosis and dysfunction.
β细胞过度凋亡与糖尿病密切相关。在糖尿病中,长期暴露于高糖环境会导致β细胞功能障碍和凋亡。刺葡萄(Vitis davidii Foex.)在传统中医中用于治疗糖尿病已有多年。在我们之前的研究中,刺葡萄籽油(TGSO)在动物模型中显示出有前景的抗糖尿病作用。然而,TGSO在抗糖尿病作用中是否发挥抗凋亡作用以及信号转导途径的相关机制尚不清楚。
用超临界二氧化碳(CO2)流体萃取法提取的TGSO处理或不处理大鼠胰腺β细胞系RIN-m5F,并通过气相色谱/质谱联用(GC/MS)进行分析。通过荧光激活细胞分选(FACS)检测细胞凋亡,采用酶联免疫吸附测定(ELISA)法检测胰岛素分泌,并通过定量逆转录聚合酶链反应(qRT-PCR)评估凋亡相关基因的表达。
TGSO含有87.02%的不饱和脂肪酸(UFA),显著减少胰腺β细胞凋亡,并保护因高糖受损的胰岛素分泌。促凋亡基因如诱导型一氧化氮合酶(iNOS)、半胱天冬酶-3(Caspase-3)、活化转录因子3(ATF-3)、应激活化蛋白激酶(JNK)、p38和Fas的表达下调,而抗凋亡基因蛋白激酶B(Akt)和Bcl-2/Bax上调。
结果表明,TGSO保护β细胞免受高糖诱导的凋亡,其保护活性可能与线粒体途径、内质网(ER)应激途径和Fas信号途径有关,这意味着TGSO可能是减少β细胞凋亡和功能障碍的一种有效的补充或替代药物。