海藻玉壶汤中海藻不同品种与甘草加减应用对甲状腺肿大模型大鼠甲状腺Bax、Bcl-2 mRNA表达的影响

Effects of different species of haizao in Haizao Yuhu Decoction with or without gancao on the expressions of Bax,Bcl-2 mRNA in rats with goiter

目的探讨含高剂量反药组合的海藻玉壶汤中海藻不同品种与甘草加减应用对甲状腺肿大模型大鼠甲状腺Bax、Bcl-2 mRNA表达的影响。方法选取Wistar大鼠144只,雌雄各半,随机分为空白组、模型组、阳性药优甲乐组(优甲乐组)、含羊栖菜的海藻玉壶汤全方组(羊栖菜全方组)、含海蒿子的海藻玉壶汤全方组(海蒿子全方组)、含羊栖菜的海藻玉壶汤去甘草组(羊栖菜去甘草组)、含海蒿子的海藻玉壶汤去甘草组(海蒿子去甘草组)、海灌玉壶汤去海藻组(全方去海藻组)、海藻玉壶汤去海藻甘草组(全方去海藻甘草组),共计9组。造模期间,除空白组外,其余各组大鼠灌服丙基硫氧嘧啶(PTU)10 mg/kg,每日1次,共给造模药14 d。造模后给予治疗药物,周期为28 d,其中,空白组与模型组给予等量蒸馏水灌胃,阳性药组灌服优甲乐,给药剂量为20μg/kg,每日1次,其余各给药组给予相应药液灌胃,大鼠每日给药1次,给药剂量为海蒿子/羊栖菜去甘草组12.6 g/kg,全方去海藻组13.5 g/kg,海蒿子/羊栖菜全方组18.0 g/kg,全方去海藻甘草组8.1 g/kg。在给药期间,为了稳定模型,除空白组外,每隔1 d,其余各组灌服PTU,剂量同前。给药结束后计算各组大鼠甲状腺系数,观察甲状腺组织病理形态,用RT-PCR法检测甲状腺组织中Bax、Bcl-2 mRNA的表达。结果与空白组比较,各组大鼠甲状腺系数明显升高(P<0.01);与模型组比较除优甲乐组外其余各给药组甲状腺系数明显降低(P<0.01)。甲状腺组织病理形态观察显示模型组与优甲乐组较空白组变化明显,甲状腺组织小叶排列紊乱,滤泡大小不一,滤泡上皮细胞弥漫性增生和肥大,其余各给药组有纠正作用,其中全方(海蒿子/羊栖菜)组组织病理形态最接近空白组。与空白组比较,模型组Bax mRNA表达降低(P<0.05);与模型组比较海藻玉壶汤全方及加减应用各组Bax mRNA表达升高(P<0.05,P<0.01)。各组Bcl-2 mRNA表达无差异。与空白组比较,模型组Bax/Bcl-2降低(P<0.05),与模型组比较,海藻玉壶汤全方及加减应用各组数值均升高(P<0.05,P<0.01)。结论含高剂量反药组合的海藻玉壶汤中不同品种海藻与甘草加减应用,对甲状腺肿大模型大鼠甲状腺系数及甲状腺组织形态有纠正作用;其作用机制可能与其升高Bax/Bcl-2从而促进甲状腺细胞凋亡相关。

Objective To investigate the effects of different species of Haizao(haihaozi, Sargassum Pallidum or yangqicai, Sargassum Fusiforme) in Haizao Yuhu(HZYH) Decoction with or without gancao(Radix Glycyrrliizae) on the expressions of Bax, Bcl-2 mRNA in goiter rats. Methods Rats(n =144) were randomly divided into 9 groups: blank group, model group, positive drug euthyrox(20 μg/kg euthyrox) group, HZYH Decoction with yangqicai without gancao group(12.6 g/kg), HZYH Decoction with haihaozi without gancao group(12.6 g/kg), HZYH Decoction without haizao group(13.5 g/kg), HZYH Decoction group with haihaozi(18.0 g/kg), HZYH Decoction group with yangqicai(18.0 g/kg), and HZYH Decoction without Haizao and Gancao group(8.1 g/kg). During the modeling period, except for the blank group, the other groups of rats were given propyl thiouracil 10 mg/kg once a day for 14 days. The investigational treatment was administered after modeling for 28 days. Among them, the blank group and the model group were given the same amount of distilled water for gavage. The rat model of goiter was induced with propyltluouracil(PTU). Except for the blank group, the others were correspondingly given interventional medicine respectively. Thyroid index of rats and pathological morphology in thyroid tissue were evaluated. The gene expressions of Bax and Bcl-2 were respectively detected. Results The levels of thyroid index in model group and eutbyrox group were significantly increased compared with blank group(P<0.01). Compared with model group, thyroid index decreased in all other groups(P<0.01). Pathological observation of thyroid tissue showed that model group and euthyrox changed significantly compared with blank group. The lobulations of thyroid gland was disordered;the follicle size was different;the thyroid follicular epithelium cells presented with diffuse hyperplasia and hypertrophy. These changes were rectified to different degrees in other groups. The histopathological morphology of HZYH Decoction with haihaozi or yangqicai group was the closest to blank group. The expression of Bax mRNA in model group was decreased compared with blank group(P<0.05). Its expressions were increased in HZYH Decoction with or without haizao and gancao groups compared with model group(P<0.05, P<0.01). There was no difference in the expression of Bcl-2 mRNA between groups. Bax/Bcl-2 in model groups was decreased compared with blank group(P<0.05). Bax/Bcl-2 were increased in HZYH Decoction with or without haizao and gancao groups compared with model group(P<0.05, P<0.01). Conclusion Different varieties of haizao in Haizao Yuhu Decoction containing high-dose antagonistic medicinal combination with modification have corrective effect on thyroid index and pathological morphology in thyroid tissue in goiter models. Its mechanism may be related with upregulation of Bax/Bcl-2, which promotes apoptosis of thyroid cells.