1. 南京中医药大学医学与生命科学学院,江苏,南京,210023
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张伟伟, 陈逸琳, 宋文婷, 等. 水提、醇提二至丸对小鼠脾脏和卵巢细胞的影响[J]. 上海中医药杂志, 2019,53(6):80-85.
ZHANG Weiwei, CHEN Yilin, SONG Wenting, et al. Effects of water and alcohol extracts of Erzhi Pills on splenic and ovarian cells in mice[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):80-85.
张伟伟, 陈逸琳, 宋文婷, 等. 水提、醇提二至丸对小鼠脾脏和卵巢细胞的影响[J]. 上海中医药杂志, 2019,53(6):80-85. DOI: 10.16305/j.1007-1334.2019.06.020.
ZHANG Weiwei, CHEN Yilin, SONG Wenting, et al. Effects of water and alcohol extracts of Erzhi Pills on splenic and ovarian cells in mice[J]. Shanghai Journal of Traditional Chinese Medicine, 2019,53(6):80-85. DOI: 10.16305/j.1007-1334.2019.06.020.
目的:探讨水提、醇提二至丸对小鼠脾脏和卵巢颗粒细胞相互作用的影响。 方法:取小鼠脾单个核细胞和卵巢颗粒细胞体外培养。①二至丸水提组在脾细胞中加入浓度为0.96 g/L的二至丸水提物,二至丸醇提组加入浓度为1.920 g/L的醇提物,阳性对照组加入浓度为0.1 g/L的香菇多糖,空白对照组加入培养液;②二至丸水提组在卵巢颗粒细胞中加入浓度为0.96 g/L的二至丸水提物,二至丸醇提组加入浓度为0.72 g/L的醇提物,阳性对照组加入浓度为0.2 g/L的香菇多糖,空白对照组加入培养液。采用MTT检测细胞增殖情况;ELISA检测培养上清中IL-1、E,2,的浓度;荧光实时定量PCR(QRT-PCR)检测IL-1受体(IL-1R)、雌激素受体(ERα)的mRNA表达水平;流式细胞术检测IL-1R、ERα的蛋白水平。 结果:①与空白对照组比较,二至丸水提物、醇提物对脾细胞、卵巢颗粒细胞均具有明显的促增殖作用(P,<,0.01),且醇提物对脾细胞的增殖作用强于水提物(P,<,0.05)。②水提、醇提二至丸均能促进卵巢颗粒细胞产生更多的E,2,(P,<,0.05),促进脾细胞产生更多的IL-1(P,<,0.01)。③与空白对照组比较,二至丸水提、醇提组脾细胞ERα的表达水平显著升高(P,<,0.01),此结果与香菇多糖组一致(P,>,0.05),且二至丸醇提物组ERα的表达水平高于二至丸水提组(P,<,0.05)。④二至丸水提组和醇提组卵巢颗粒细胞表面IL-1R的表达水平显著升高(P,<,0.05),与香菇多糖效果一样(P,>,0.05),但两者之间差异无统计学意义(P,>,0.05)。 结论:二至丸水提物、醇提物可通过直接调控卵巢颗粒细胞和脾细胞对小鼠生殖内分泌和免疫系统发挥良好的双向促进作用,且二至丸醇提物的效果优于水提物。
Objective:To explore the effects of water and alcohol extracts of Erzhi Pills on splenic cells and ovarian granulose cells in mice. MethodsThe splenic mononuclear cells and ovarian granulosa cells were cultured in vitro. ①Water extract of Erzhi Pills at a concentration of 0.96 g/L was added to the splenic cells in the water extract group, alcohol extract of Erzhi Pills at a concentration of 1.920 g/L in the alcohol extract group, lentinan at a concentration of 0.1 g/L in the positive control group and culture medium in the blank control group. ②Water extract of Erzhi Pills at a concentration of 0.96 g/L was added to the ovarian granulose cells in the water extract group, alcohol extract of Erzhi Pills at a concentration of 0.72 g/L in the alcohol extract group, lentinan at a concentration of 0.2 g/L in the positive control group and culture medium in the blank control group. Cell proliferation was detected by MTT; ELISA was used to determine the concentrations of IL-1 and E,2, in the culture supernatant; fluorescence quantitative real-time PCR (QRT-PCR) was used to detect mRNA expression of IL-1 receptor (IL-1R) and estradiol receptor (ERα); the levels of IL-1R and ERα were detected by flow cytometry. Results:①Compared with the blank control group, water and alcohol extracts of Erzhi Pills promoted proliferation of the splenic cells and ovarian granulose cells significantly (P,<,0.01), and alcohol extract showed a stronger effect than water extract in promoting proliferation of the splenic cells (P,<,0.05). ②Water and alcohol extracts of Erzhi Pills promoted the ovarian granulose cells to produce more E,2, (P,<,0.05) and the splenic cells to produce more IL-1 (P,<,0.01). ③Compared with the blank control group, the expression level of ERα in the splenic cells increased significantly in the water and alcohol extract groups (P,<,0.01), which was consistent with the results in the lentinan group (P,>,0.05), and the expression level of ERα in the alcohol extract was higher than that in the water extract group (P,<,0.05). ④The expression of IL-1R on the surface of ovarian granulose cells increased significantly both in the water and alcohol extract groups (P,<,0.05), consistent with the results in the lentinan group (P,>,0.05), but there were no statistically significant differences between them (P,>,0.05). Conclusion:The water and alcohol extracts of Erzhi Pills have a good bi-directional promoting effect on the reproductive, endocrine and immune systems of mice through direct regulation of ovarian granulose cells and splenic cells, and the effect of alcohol extract was better than that of water extract.
脾细胞卵巢颗粒细胞细胞培养二至丸水提醇提
splenic cellsovarian granulose cellscell cultureErzhi Pillswater extractalcohol extract
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