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目的探讨大黄及大黄蒽醌类有效成分对胰腺腺泡细胞AR42J损伤的保护作用。方法取生长良好的AR42J细胞,以1×105·mL-1浓度接种于6孔板中,设空白对照组、模型组、大黄含药血清(10%,5%,2.5%,1.25%,0.625%)组,每组3个复孔,培养24 h待细胞贴壁后弃上清,空白对照组加入含10%空白血清培养液、模型组用含10%空白血清培养液配置造模液(雨蛙素终浓度为10-7mol·L-1,脂多糖终浓度为10 mg·L-1)、大黄含药血清组分别加入不同浓度的含药血清,继续培养24 h后,检测细胞上清液中淀粉酶含量,取细胞检测细胞内胰蛋白酶、脂肪酶的含量。另取AR42J细胞,以1×105·mL-1浓度接种于6孔板中,设正常对照组、模型组、大黄素、大黄酸、芦荟大黄素、大黄酚、大黄素甲醚组,上述大黄蒽醌类成分分别设10,20,40,80滋mol·L-1共4个剂量,每组3个复孔,在37℃、5%CO2条件下,分别培养3,6,12,24 h后,取细胞上清液,检测上清液中淀粉酶的含量。结果大黄含药血清在2.5%到10%浓度范围内,能显著抑制AR42J细胞内蛋白酶、脂肪酶含量,降低细胞培养上清液中淀粉酶的水平。对大黄蒽醌的研究结果提示,大黄素在模型复制3 h后即能显著降低AR42J细胞分泌淀粉酶,且呈明显的剂量依赖性,在24 h内,大黄素各剂量组均表现出较强的抑制淀粉酶的作用;芦荟大黄素及大黄酸在40和80滋mol·L-1剂量下也能显著降低细胞上清液中淀粉酶水平,而大黄素甲醚及大黄酚作用较弱。结论大黄蒽醌是大黄保护胰腺腺泡细胞损伤的重要物质基础,其中,大黄素的作用最强。
Objective To investigate the protective effect of the active components of rhubarb and anthraquinones on the injury of pancreatic acinar cells AR42J. Methods AR42J cells with good growth were inoculated into 6-well plates at the concentration of 1 × 105 · mL-1. The blank control group, model group, rhubarb serum containing 10%, 5%, 2.5%, 1.25%, 0.625 %) Group. Each group of three replicate wells were cultured 24 h until the cells adherent to the supernatant, the blank control group was added with 10% blank serum culture medium, the model group with 10% blank serum culture solution configuration fluid ( The final concentration of rabegin was 10-7mol·L-1, the final concentration of lipopolysaccharide was 10 mg · L-1). The rhubarb serum-containing groups were added with different concentrations of serum containing drugs, cultured for 24 h, the detection of cell supernatant Liquid amylase content, take cells to detect intracellular trypsin, lipase content. In addition, AR42J cells were inoculated into 6-well plates at the concentration of 1 × 105 · mL-1. The normal control group, model group, emodin, rhein, aloe-emodin, chrysophanol, The anthraquinones were divided into 4 doses of 10, 20, 40, and 80 mol L-1 respectively. Three replicate wells of each group were cultured at 37 ℃ and 5% CO2 for 3, 6, 12 and 24 After h, the cell supernatant was taken and the amount of amylase in the supernatant was measured. Results Rhubarb serum containing 2.5% to 10% concentration range, can significantly inhibit AR42J intracellular protease, lipase content, reduce the level of amylase in cell culture supernatant. The results of anthraquinone on rhubarb suggest that emodin can significantly reduce the amylase secretion in AR42J cells 3 h after model replication, and shows a dose-dependent manner. Emodin in each dose group showed a strong effect within 24 h Of the inhibition of amylase; aloe-emodin and rhein at 40 and 80? L -1 dose can significantly reduce the level of amylase in the cell supernatant, and the role of macrolide and chrysophanol weak. Conclusion Rhubarb anthraquinone is an important material basis for rhubarb to protect pancreatic acinar cells. Among them, emodin has the strongest effect.