大黄及大黄蒽醌类有效成分对胰腺腺泡细胞损伤的保护作用

Protective Effects of Radix et Rhizoma Rhei and Its Anthraquinone Effective Components on Pancreatic Acinar Cell Injury

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 effects of Radix et Rhizoma Rhei （RRR） and its anthraquinone effective components on injury of pancreatic acinar AR42J cells. Methods Rat pancreatic acinar AR42J cells were cultured in 6-well plates at the concentration of 1 ×10^5·mL^-1, and were divided into normal control group（ 10 % blank serum） , cute pancreatitis （AP） model group （10 % blank serum culture with 10-7 mol·L^-1 of cerulein and 10 mg·L^-1 of lipoplysaeeharide added）, RRR-containing serum（10 %, 5 %, 2.5 %, 1.25 %, 0.625 % medicated seruum） groups. After culturing for 24 continuous hours, the amylase content in the cell supernatant, and the cellular trypsin and lipase were measured. In the other experiment, AR42J cells were cultured in 6-well plates at the concentration of 1 ×10^5 ·mL^-1, and were divided into normal control group model group, rhubarb anthraquinone groups（emodin, rhein, aloe-emodin, ehrysophanol, and physcion at final concentration of 10, 20, 40, 80 μmol·L^-1, respectively）. After culturing for 3, 6, 12, 24 hours under the conditions of 37 ℃ and 5 % C02, the level of amylase in cell supernatant was measured. Results Within the dose range of 2.5 % to 10 %, RRR-containing serum could inhibit the content of the trypsin and lipase in AR42J cells significantly, and at the same time, could decrease the level of amylase in the supernatant of cell culture fluid. The results of rhubarb anthraquinone showed that emodin couldsignificantly decrease the secretion of amylase in AR42J cells after modeling for 3h, presenting an obvious dose-dependent manner. Within the limits of 24h, emodin at different concentrations showed strong inhibitory effects on amylase levels. Aloe-emodin and rhein at the concentrations of 40 and 80 μmol· L^-1 could also significantly reduce amylase levels in the cell supernatant, but the effects of the physcion and chrysophanol were weak. Conclusion Anthraquinone is the important material of Radix et Rhizoma Rhei which could protect pancreatic acinar cells from damage, and the protective effect of emodin is the strongest.