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目的:研究雷公藤内酯醇药物血清对体外培养的大鼠肾上腺皮质细胞分泌功能的影响。方法:将SD大鼠30只随机分成对照组、泼尼松组和低、中、高剂量雷公藤内酯醇组,相应药物灌胃7d制备药物血清。分离正常大鼠肾上腺皮质细胞,分别加入药物血清常规培养48h后,采用免疫组织化学法观察增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达,并计算PCNA阳性细胞数;透射电子显微镜观察肾上腺皮质细胞的超微结构;酶联免疫吸附测定法检测细胞培养上清液中皮质酮水平;实时荧光定量聚合酶链式反应法检测肾上腺皮质细胞3β-羟基类固醇脱氢酶(3β-hydroxysteroi ddehydrogenase,3β-HSD)mRNA表达水平。结果:各个浓度雷公藤内酯醇药物血清组肾上腺皮质细胞培养上清液中皮质酮水平及细胞中3β-HSDmRNA表达水平较对照组明显升高;中、高剂量雷公藤内酯醇组PCNA阳性细胞比率高于对照组(P<0.05),泼尼松组上清液中皮质酮水平、细胞中3β-HSDmRNA表达水平和PCNA阳性细胞比率均低于对照组(P<0.05)。结论:雷公藤内酯醇对体外培养的大鼠肾上腺皮质细胞有刺激作用,可以促使肾上腺皮质细胞分泌皮质酮。
Objective: To study the effects of triptolide serum on secretion of adrenocortical cells in vitro. Methods: Thirty SD rats were randomly divided into control group, prednisone group and low, medium and high doses of triptolide group. The corresponding drugs were administrated for 7 days to prepare drug serum. Adrenal cortical cells were isolated from normal rats. The serum was cultured for 48 hours. The proliferating cell nuclear antigen (PCNA) expression was detected by immunohistochemistry and the number of PCNA positive cells was counted. The adrenal cortex The ultrastructure of the cells was detected by enzyme-linked immunosorbent assay (ELISA). The level of corticosterone in the cell culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The 3β-hydroxysteroid dehydrogenase -HSD) mRNA expression level. Results: The levels of corticosterone and the level of 3β-HSD mRNA in adrenal cortical cell culture supernatant of triptolide drug serum group were significantly higher than those in control group. The ratio of PCNA positive cells in medium and high dose triptolide group The levels of corticosterone, 3β-HSD mRNA and PCNA positive cells in the supernatant of prednisone group were significantly lower than those in the control group (P <0.05). Conclusion: Triptolide stimulates rat adrenocortical cells cultured in vitro and can induce adrenocortical cells to secrete corticosterone.