目的 研究地黄、知母、黄柏单独及配伍使用对药源性阴虚证小鼠肾上腺皮质功能的保护作用。方法 72只雄性ICR小鼠随机分为对照组、模型组、地黄组、知母组、黄柏组、地黄-知母-黄柏组，每组12只。采用25 mg/（kg·d）氢化可的松ig给予小鼠，每日1次，连续5 d，复制药源性阴虚证模型，给药组分别ig给予地黄[9.0 g/（kg·d）]、知母[（6.0 g/（kg·d）]、黄柏[（5.0 g/（kg·d）]、地黄-知母-黄柏配伍[（6.7 g/（kg·d）]的水煎药液，连续给药5 d。透射电子显微镜检测肾上腺皮质超微结构；ELISA检测血清皮质酮；实时荧光定量PCR技术检测肾上腺类固醇激素合成酶基因（Star、Cyp11a1、Cyp21a1、Cyp11b1）、胆固醇摄取基因（Ldlr；Scarb1，蛋白名SRB1）、胆固醇合成限速酶基因（Hmgcr）、胆固醇储存基因（Acat1、Lipe）、胆固醇流出基因（Abca1、Abcg1）、胆固醇稳态基因（Nr1h3，蛋白名LXRα）表达变化；Western blotting检测肾上腺LDLR、SRB1、LIPE、ACAT1、CYP11A1、StAR、LXRα蛋白表达。结果 与对照组比较，模型组小鼠体质量、血清皮质酮水平均显著下降（P<0.05），肾上腺束状带细胞出现典型的脂滴融合现象、线粒体萎缩；氢化可的松显著抑制肾上腺皮质激素合成过程Star、Cyp21a1、Cyp11b1、Ldlr、Scarb1、Acat1基因表达（P<0.05），以及抑制StAR、LDLR、SBR1和LXRα蛋白表达，促进Abcg1基因表达（P<0.05）。与模型组比较，各药物治疗后肾上腺束状带细胞未见明显的脂滴融合；地黄、知母、黄柏单独治疗后显著抑制皮质酮分泌（P<0.05）；地黄-知母-黄柏配伍显著提升Star、Cyp11a1、Cyp21a1、Lipe、Abca1、Nr1h3基因表达（P<0.05），并增强CYP11A1、StAR、LDLR、SBR1和LXRα蛋白表达。结论 地黄、知母、黄柏单独给药及其配伍治疗可通过调节胆固醇稳态平衡及恢复部分受抑制的皮质激素合成酶，配伍给药更有助于改善受损伤的肾上腺皮质功能。

Objective To study the effects of Rehmanniae Radix (RR), Anemarrhenae Rhizoma (AR) and Phellodendri Chinensis Cortex (PCC) on adrenal function in mice with drug-induced yin deficiency syndrome. Methods A total of 72 male ICR mice were randomly divided into normal control group, hydrocortisone model group, RR treatment group, AR treatment group, PCC treatment group, and RR-AR-PCC compatibility treatment group, with12 mice in each group. The mice were given 25 mg/(kg·d) hydrocortisone by intragastric administration once a day for 5 consecutive days to replicate the drug-induced yin deficiency syndrome model, and then administrated 9 g/(kg·d) RR, 6.0 g/(kg·d) AR, 5.0 g/(kg·d) PCC, 6.7 g/(kg·d) RR-AR-PCC compatibility of water decoction liquid for 5 d. The adrenal cortex was examined by transmission electron microscopy; Serum corticosterone were measured by ELISA; The expressions of Star, Cyp11a1, Cyp21a1, Cyp11b1, Ldlr, Scarb1, Hmgcr, Acat1, Lipe, Abca1, Abcg1, Nr1h3 genes were detected by real-time quantitative PCR. The expressions of LDLR, SRB1, LIPE, ACAT1, CYP11A1, StAR, LXRα protein were detected by Western blotting. Results Compared with the normal control group, the body weight and serum corticosterone levels of the mice were significantly decreased after 5 d of administration of 25 mg/(kg·d) hydrocortisone (P < 0.05). Adrenal gland zona fasciculate cells appeared fat droplet fusion and mitochondrial atrophy. Hydrocortisone significantly inhibited the expression of Star, Cyp21a1, Cyp11b1, Ldlr, Scarb1 and Acat1 genes expression in adrenocortical hormone synthesis (P < 0.05), and inhibited the expression of StAR, LDLR, SBR1 and LXRα proteins (P < 0.05), and promoted the Abcg1 gene expression (P < 0.05). Compared with the model group, no obvious lipid droplet fusion was observed in the adrenal gland zona fasciculate cells after each drug treatment; RR, AR and PCC significantly inhibit corticosterone secretion after treatment alone (P < 0.05); The combination of RR, AR and PCC significantly increased the expression of Star, Cyp11a1, Cyp21a1, Lipe, Abca1, Nr1h3 genes (P < 0.05), and enhanced the expression of CYP11A1, StAR, LDLR, SBR1 and LXRα proteins (P < 0.05). Conclusion RR, AR and PCC combination can improve the damaged adrenal function more than their alone by regulating cholesterol homeostasis and restoring partially inhibited corticosteroid synthase.

R285.5

国家自然科学基金面上项目（81873212）；上海市科委项目（19140905000）