标题：灵芝提取物及其含药血清对胰岛β细胞的影响

      作者：张强，杨宏杰，何燕铭，张曾    (上海中医药大学附属岳阳中西医结合医院内分泌科，上海 200437)

      卷次： 2018年29卷9期

      【摘要】 目的 研究灵芝提取物灵芝蛋白多糖(FYGL)对胰岛β细胞是否有保护作用。方法 (1)配置不同浓度的FYGL (0.01 mg/mL、0.05 mg/mL、0.1 mg/mL)，培养min6细胞株，接种于96孔版，细胞贴壁后加入上述不同剂量FYGL干预72 h，XTT法检测各组细胞活性；(2)中药血清药理学方法制备大鼠含药血清。15只健康SPF级Wistar雄性大鼠，随机分成五组，分别为对照组(磷酸盐缓冲液)、西格列汀2 mg/kg组、FYGL低剂量组(75 mg/kg)、FYGL中剂量组(250 mg/kg)、FYGL高剂量组(450 mg/kg)，每组3只，连续10 d灌胃给药后处死取血清，同组3只血清合并备后续实验用；同上方法培养min6细胞株，加入含有10%上述含药血清的DMEM培养液干预72 h，XTT法检测各组细胞活性；(3)同上方法培养min6细胞株，加入不同浓度(50 μmol/L、100 μmol/L、250 μmol/L、500 μmol/L)棕榈酸(PA)作用72 h，XTT法检测各组细胞活性；(4)同上方法培养min6细胞株，加入400 μmol/L棕榈酸(PA)及10%上述含药血清的DMEM培养液共同干预72 h，XTT法检测各组细胞活性。结果 (1)与对照组[(100±5.62)%]相比，FYGL呈剂量依赖性促进min6细胞增殖[(131.14±12.53)%、(174.48±17.09)%、(259.28±9.51%)%]，差异均有显著统计学意义(P<0.01)；(2)与对照组[(100±10.14)%]相比，西格列汀及 FYGL含药血清呈剂量依赖性促进min6细胞增殖[(119.27±5.27)%、(112.5±24.15)%、(125.65±1.52)%、(133.98±9.95)%]，西格列汀及中高剂量FYGL组比较差异均有显著统计学意义(P<0.01)；(3)与对照组[(100±9.26)%]相比，250 μmol/L及500 μmol/L PA可诱导min6细胞凋亡[(75.64±7.74)%、(53.63±9.61)%]，差异均有显著统计学意义(P<0.01)；(4)与对照组相比，400 μmol/L PA可明显促进min6细胞凋亡[(100±5.76)% vs (56.89±1.86)%]，差异有显著统计学意义(P<0.01)；与 PBS血清组相比，西格列汀含药血清组及FYGL含药血清组可抑制 PA导致的min6细胞凋亡，且以 FYGL 250 mg/kg作用最显著[(56.89±1.86%) vs (88.53±24.84)%]，差异均有统计学意义(P<0.01)。结论 灵芝提取物FYGL促进min6细胞增殖且能抑制PA诱导min6细胞凋亡作用，可能作为一种具有保护胰岛β细胞作用的潜在降糖药物。
      【关键词】 2型糖尿病；灵芝提取物FYGL；min6细胞；细胞凋亡
      【中图分类号】 R587.1 【文献标识码】 A 【文章编号】 1003—6350（2018）09—1188—04

Effects of Ganoderma lucidum extract and the extract-containing serum on islet β cells.
ZHANG Qiang, YANGHong-jie, HE Yan-ming, ZHANG Zeng. Department of Endocrinology, Yueyang Hospital of Integrated Traditional Chineseand Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, CHINA
【Abstract】 Objective To explore whether Ganoderma lucidum extract Ganoderma lucidum polysaccharide(FYGL) had protective effects on islet β cells. Methods (1) Different concentrations of FYGL (0.01 mg/mL, 0.05 mg/mL,0.1 mg/mL) were used to cultivate min6 cell lines in 96-well plates. After the cells were adhered, the above different dosesof FYGL were added for the intervention of 72 hours, and XTT assay was used to detect the cell viability. (2) Serum Phar-macology of Chinese Medicine was applied to prepare FYGL-containing rat serum. Fifteen healthy SPF Wistar male ratswere randomly divided into 5 groups: control group (phosphate buffer saline, PBS), sitagliptin 2 mg/kg group, FYGL lowdose group (75 mg/kg), FYGL middle dose group (250 mg/kg), and FYGL high dose group (450 mg/kg), with three rats ineach group. The rats were sacrificed 10 days after gavage. Serum was collected from the rats in the same group. The min6cell line was cultured in the same manner as above, and the cells were added with DMEM medium with FYGL-containingserum (10%) for 72 hours. XTT assay was used to detect the cell viability. (3) The min6 cell line was cultured in the samemanner as above, and cells were treated with different concentrations (50 μmol/L, 100 μmol/L, 250 μmol/L, 500 μmol/L)of palmitic acid (PA) for 72 hours. XTT assay was used to detect the cell viability. (4) The min6 cell line was cultured inthe same manner as above, and cells were incubated with 400 μmol/L PA and DMEM medium with FYGL-containing se-rum (10%) for 72 hours. XTT assay was used to detect the cell viability. Results (1) Compared with the control group(100±5.62)%, FYGL significantly promoted the proliferation of min6 cells in a dose-dependent manner: (131.14±12.53)%in FYGL low dose group, (174.48±17.09)% in FYGL middle dose group, (259.28±9.51) in FYGL high dose group (P<0.01). (2) Compared with the control group (100±10.14)%, sitagliptin and FYGL-containing serum increased the prolifera-tion of min6 cells in a dose-dependent manner: (119.27±5.27)%, (112.5±24.15)%, (125.65±1.52)%, (133.98±9.95%), andthere were significant differences between sitagliptin and medium and high dose FYGL group (P<0.01). (3) Comparedwith the control group (100±9.26)%, 250 μmol/L and 500 μmol/L PA could significantly induce the apoptosis of min6cells: (75.64±7.74)%, (53.63±9.61)%, P<0.01. (4) Com