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补肾活血化痰方改善雄激素致不孕大鼠高雄激素血症的机制探讨
俞瑾1△,张洁2,3△,韩洁2,雷玲2,俞超芹2*,翟东霞2*
0
(1. 上海中医药大学,上海 201203
2. 第二军医大学长海医院中医科,上海 200433
共同第一作者
*通信作者)
摘要:
目的 观察补肾活血化痰方(BHHR)对雄激素致不孕大鼠(ASR)高雄激素血症的治疗作用及治疗前后ASR卵巢内雄激素合成酶和代谢酶的表达量的变化,并探讨其可能的作用机制。方法 SD雌性大鼠于9日龄在颈背部皮下一次性注射丙酸睾丸酮(1.25 mg)制备ASR模型。将造模成功的ASR随机分为3组,分别为模型组(蒸馏水10 mL/kg灌胃)、二甲双胍治疗组(0.1 g/kg灌胃)和BHHR治疗组(10 mL/kg灌胃),每组各13只。另设正常对照组(n=10)。疗程(28 d)结束后观察大鼠的性周期恢复情况、体质量及卵巢质量/体质量比值。采用放射免疫分析法测定血清睾酮(T)水平;免疫组织化学定量方法检测卵巢3β-羟甾脱氢酶、细胞色素P450 17α-羟化酶/17,20-裂解酶和P450芳香化酶的表达量。结果 (1)性周期恢复:BHHR治疗组及二甲双胍治疗组中恢复性周期的大鼠数量多于模型组(P<0.01)。 (2)体质量:模型组大鼠体质量高于正常对照组(P<0.01);BHHR治疗组与二甲双胍治疗组大鼠体质量均较模型组降低(P<0.01)。(3)卵巢质量/体质量比值:模型组大鼠卵巢质量/体质量比值高于正常对照组、BHHR治疗组及二甲双胍治疗组(P<0.01)。(4)血清T水平:模型组大鼠血清T值高于正常对照组、BHHR治疗组及二甲双胍治疗组(P<0.01)。(5)免疫组织化学结果:与模型组和正常对照组相比,BHHR治疗组大鼠P450arom表达升高 (P<0.01);BHHR治疗组3β-羟甾脱氢酶、细胞色素P450 17α-羟化酶/17,20-裂解酶的表达与模型组间差异无统计学意义(P>0.05)。结论 BHHR可以降低ASR模型大鼠血清T水平,其机制可能是通过上调雄激素代谢酶P450arom的表达而实现的。
关键词:  补肾活血化痰方  雄激素增多症  3β-羟甾脱氢酶  类固醇17α-羟化酶  P450芳香化酶
DOI:10.3724/SP.J.1008.2013.00498
投稿时间:2013-02-20修订日期:2013-03-27
基金项目:国家自然科学基金(30371838,30672742, 30873353), 国家自然科学基金重点项目(30930113), 国家自然科学基金青年科学基金(81001535).
Mechanism of Bushen huoxue huatan recipe improving hyperandrogenism in androgen-induced sterile rats
YU Jin1△,ZHANG Jie2,3△,HAN Jie2,LEI Ling2,YU Chao-qin2*,ZHAI Dong-xia2*
(1. Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2. Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
Co-first authors.
*Corresponding authors.)
Abstract:
Objective To observe the therapeutic effect of Bushen huoxue huatan recipe (BHHR) on androgen-induced sterilized rats (ASR) and the expression of androgen synthase and metabolic enzymes in the ovary of ASR before and after treatment with BHHR, and to discuss the possible mechanism. Methods Female SD rats of 9-day old were injected subcutaneously with testosterone propionate (1.25 mg) to create model. The model rats were randomly divided into 3 groups: model group (treated with distilled water by gastrogavage, 10 mL/kg), metformin therapy group (gastrogavage, 0.1 g/kg) and BHHR therapy group (gastrogavage, 10 mL/kg), with 13 animals in each group. Ten rats with normal estrous cycle served as normal controls. The body mass, sexual cycle recovery and ovary mass/body mass ratio were observed after 28-day treatment. Serum testosterone level was measured by radioimmunoassay; the expressions of 3β-hydroxylsteroid dehyrogenase (3β-HSD), cytochrome P450 17α-hydroxylase/17, 20-lyase (CYP17) and P450 aromatase (P450arom) in ovary were detected by quantitative immunohistochemistry method. Results (1) Significantly more rats in the metformin and BHHR groups had sexual cycle recovery compared with that in the model group (P<0.01). (2) The body mass of rats in the model group was significantly heavier than that in the normal control group (P<0.01), and those of the BHHR and metformin groups were significantly lighter than that of ASR model group (P<0.01). (3) Ovary mass/body mass ratio of the model rats was significantly higher than those of the other three groups (P<0.01). (4) Serum testosterone level in the model group was significantly higher than those in the other three groups (P<0.01). (5) Compared with the model group and the normal group, the P450arom expression in the BHHR group was significantly increased (P<0.01), while no significant difference was found in 3β-HSD and CYP17 expression between the BHHR group and the model group (P>0.05). Conclusion BHHR can reduce serum testosterone levels in ASR rat, which might be through up-regulating the expression of the androgen metabolism enzyme P450arom.
Key words:  Bushen huoxue huatan recipe  hyperandrogenism  3beta-hydroxysteroid dehydrogenase  steroid 17alpha-hydroxylase  cytochrome P450 aromatase