海藻玉壶汤加减海藻甘草反药组合对甲状腺肿大大鼠p53/p21/Caspase-3通路表达的影响  

Effect of Haizao Yuhutang Combined with Modified Sargassum and Glycyrrhizae Radix et Rhizoma on Expression of p53/p21/Caspase-3 Pathway in Rats with Goiter

作　　者：田颐 董肖 刘晓庆 曹灿[1] 吴美晶 徐向楠 廖文勇 陈绍红[1] 柳海艳[1] 于雪[1] 范盎然[1] 修琳琳[1] 钟赣生[1] TIAN Yi;DONG Xiao;LIU Xiaoqing;CAO Can;WU Meijing;XU Xiangnan;LIAO Wenyong;CHEN Shaohong;LIU Haiyan;YU Xue;FAN Angran;XIU Linlin;ZHONG Gansheng(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2024年第16期68-75,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274118)。

摘　　要：目的:探究海藻和甘草反药组合在2020年版《中华人民共和国药典》高限2倍剂量条件下于海藻玉壶汤中加减配伍应用对治疗甲状腺肿大的作用和p53/p21/胱天蛋白酶-3(Caspase-3)信号通路的影响。方法:Wistar大鼠128只,随机分为空白组、模型组、优甲乐组(20μg·kg^(-1))、海藻玉壶汤组(12.06 g·kg^(-1))、去海藻组(9.90 g·kg^(-1))、去甘草组(10.26 g·kg^(-1))、去海藻甘草组(8.10 g·kg^(-1))、海藻甘草组(3.96 g·kg^(-1)),每组16只。除空白组其余各组给予PTU复制甲状腺肿大病理模型,将优甲乐用作阳性药,其余各组给予相应药液,末次给药后12 h进行取材。观察各组甲状腺形态;计算各组大鼠甲状腺系数;检测各组大鼠血清中三碘甲状腺原氨酸(T3)、甲状腺素(T4)、游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺激素(TSH)的含量;苏木素-伊红(HE)染色观察甲状腺病理变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测甲状腺组织p53、p21、p38丝裂原活化蛋白激酶(p38 MAPK)、细胞周期蛋白D_(1)(Cyclin D_(1))、Caspase-3 mRNA表达情况;蛋白免疫印迹法(Western blot)检测甲状腺组织凋亡相关信号通路p53、p21、Caspase-3蛋白表达情况。结果:与空白组比较,模型组甲状腺系数显著增高(P<0.01),T3、T4、FT3、FT4水平均降低(P<0.01),TSH升高(P<0.01),甲状腺组织病理形态变化明显(P<0.05),p53、p21、Cyclin D_(1)、Caspase-3 mRNA表达均降低(P<0.05);与模型组比较,全方组甲状腺系数显著降低(P<0.01),T3、T4、FT3、FT4水平均上升(P<0.05),TSH下降(P<0.01),甲状腺组织病理形态明显恢复(P<0.05),p53、p21、Caspase-3 mRNA表达上升(P<0.05),p53、p21、Caspase-3蛋白表达量增加(P<0.05)。结论:在2020年版《中华人民共和国药典》高限2倍剂量条件下,含海藻和甘草反药组合于海藻玉壶汤中加减配伍应用对甲状腺肿大模型大鼠的甲状腺系数、甲状腺激素及甲状腺组织�Objective:To investigate the effect of the modified application of Sargassum and Glycyrrhizae Radix et Rhizoma combination that was two times the dose of the high limit in the Pharmacopoeia of the People's Republic of China 2020 in Haizao Yuhutang(HYT)on the goiter and tumor suppressor protein(p53),cell cycle-dependent protein kinase inhibitor(p21),and p53/p21/Caspase-3 signaling pathway.Method:A total of 128 Wistar rats were randomly divided into blank group,model group,euthyrox group(20μg·kg^(-1)),HYT group(12.06 g·kg^(-1)),group without Sargassum(9.90 g·kg^(-1)),group without Glycyrrhizae Radix et Rhizoma(10.26 g·kg^(-1)),group without Sargassum and Glycyrrhizae Radix et Rhizoma(8.10 g·kg^(-1)),and group with Sargassum and Glycyrrhizae Radix et Rhizoma(3.96 g·kg^(-1)),with 16 rats in each group.All groups,except for the blank group,were given PTU to replicate the pathology model of goiter,with euthyrox as the positive drug,and the other groups were given the corresponding medicinal solution.The sample was collected 12 hours after the last administration.The morphology of the thyroid in each group was observed,and the thyroid coefficients of rats in each group were calculated.The levels of triiodothyronine(T3),thyroxine(T4),free triiodothyronine(FT3),free thyroxine(FT4),and thyroid-stimulating hormone(TSH)were detected in the serum of the rats in each group.The pathological changes in the thyroid were observed by hematoxylin-eosin(HE)staining.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expressions of p53,p21,and p38 mitogen-activated protein kinase(p38 MAPK),Cyclin D_(1),and Caspase-3 in thyroid tissue.Western blot was used to detect the protein expression of p53,p21,and Caspase-3 in apoptosis-related signaling pathways in thyroid tissue.Result:Compared with the blank group,the thyroid coefficient of the model group was significantly higher(P<0.01),and the levels of T3,T4,FT3,and FT4 were all reduced(P<0.01).The level of TSH was increased(P<0.01

关 键 词：海藻玉壶汤 十八反 甲状腺肿模型大鼠 凋亡 p53 P21 胱天蛋白酶-3(Caspase-3) 

分 类 号：R2-0[医药卫生—中医学] R33R289R581