摘要
目的:通过观察消瘰丸对实验性甲状腺肿大鼠磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白复合物1(m TORC1)通路的影响,探究其对实验性甲状腺肿大鼠的治疗效果。方法:购买60只5月龄SPF级SD大鼠,雌雄各半,其中10只作为正常组,其余大鼠灌服丙基硫氧嘧啶(PTU)溶液构建结节性甲状腺肿大鼠模型,建模成功后随机分为模型组、左甲状腺素钠片组、消瘰丸低、中、高剂量组各10只。左甲状腺素钠片组采用15μg·kg^(-1)的左甲状腺素钠片灌胃,消瘰丸低、中、高剂量组分别采用低剂量10 g·kg^(-1)、中剂量20 g·kg^(-1)、高剂量30 g·kg^(-1)消瘰丸灌胃,正常组、模型组采用等体积0.9%氯化钠溶液灌胃。干预4周后采用5%苯巴比妥做常规腹腔注射麻醉后处死,显微镜观察大鼠病理组织学、采用罗氏电化学发光免疫分析仪检测血清甲状腺激素水平、酶联免疫吸附测定法(ELISA)检测血清细胞生长因子、采用高效液相色谱仪检测神经递质、蛋白免疫印迹法(Western blot)检测PI3K/Akt/m TORC1信号通路蛋白表达量。结果:与正常组比较,模型组碱性成纤维生长因子(b FGF)、血管内皮生长因子(VEGF)、胰岛素样生长因子-1(IGF-1)、5-羟色胺(5-HT)、促甲状腺激素(TSH)水平、PI3K、Akt、m TORC1蛋白表达量升高,去甲肾上腺素(NE)、三碘甲腺原氨酸(T_(3))、四碘甲腺原氨酸(T_(4))、游离三碘甲状腺原氨酸(FT_(3))、游离甲状腺激素(FT_(4))水平降低(P<0.05);与模型组比较,左甲状腺素钠片组、消瘰丸低、中、高剂量组b FGF、VEGF、IGF-1、5-HT、TSH水平,PI3K、Akt、m TORC1蛋白表达量降低,NE、T_(3)、T_(4)、FT_(3)、FT_(4)水平升高(P<0.05);左甲状腺素钠片组比较,消瘰丸低、中、高剂量组b FGF、VEGF、IGF-1、5-HT、TSH水平,PI3K、Akt、m TORC1蛋白表达量降低,NE、T_(3)、T_(4)、FT_(3)、FT_(4)水平升高(P<0.05)。结论:消瘰丸可能作用于PI3K/Akt/m TORC1信号通路,发挥其治疗结节性甲状腺肿的作用,且表现为剂量依赖。
Objective:By observing the effect of Xiaoluowan on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin complex 1(m TORC1)pathway in experimental goiter rats,this study aims to explore its therapeutic effect on experimental goiter rats.Method:Sixty 5-month-old SDrats of SPF grade were purchased,half males and half females,of which 10 were used as a normal group,and the remaining rats were administrated with propylthiouracil(PTU)solution to induce nodular goiter.After successful modeling,rats were randomly divided into a model group,levothyroxine sodium tablets group,Xiaoluowan low-dose group,medium-dose group,and high-dose group,ten rats each.The levothyroxine sodium tablets group was given 15μg·kg^(-1)levothyroxine sodium tablets by gavage.The Xiaoluowan low-,medium-,and high-dose groups were given(ig)Xiaoluowan low-dose(10 g·kg^(-1)),medium-dose(20 g·kg^(-1)),and high-dose(30 g·kg^(-1))Xiaoluowan,and the normal group and model group were administered(ig)with the same volume of 0.9%sodium chloride solution.Four weeks after the intervention,rats were sacrificed by routine intraperitoneal anesthesia using 5%phenobarbital.Subsequently,the histopathology was observed under a microscope,and serum thyroid hormone levels were measured using a Roche electrochemiluminescence immunoassay analyzer.Serum cytokines were detected by enzyme-linked immunosorbent assay(ELISA),and neurotransmitters were measured using a high-performance liquid chromatograph.The protein level of PI3K/Akt/m TORC1 pathway was determined by Western blot.Result:As compared with the normal group,the levels of basic fibroblast growth factor(b FGF),vascular endothelial growth factor(VEGF),insulin-like growth factor(IGF-1),5-hydroxytryptamine(5-HT),and thyroid stimulating hormone(TSH)were increased,and PI3K,Akt,and m TORC1 protein levels were up-regulated in the model group,while the levels of norepinephrine(NE),triiodothyronine(T_(3)),tetraiodothyronine(T_(4)),free triiodothyronine(FT_(3)),and free thyroid hormone(FT_(4))were decreased(P<0.05).Compared with the model group,the levothyroxine sodium tablets group,and Xiaoluowan low-,medium-,and high-dose groups exhibited reduced levels of b FGF,VEGF,IGF-1,5-HT,and TSH,and down-regulated PI3K,Akt,and m TORC1 protein levels,and increased NE,T_(3),T_(4),FT_(3),and FT_(4)levels(P<0.05).Conclusion:Xiaoluowan may act on the PI3K/Akt/m TORC1 signaling pathway to play its role in the treatment of nodular goiter,and it is dose-dependent.
作者
梁伟
孙禹
陈丽新
程伟
李文兰
LIANG Wei;SUN Yu;CHEN Li-xin;CHENG Wei;LI Wen-lan(School of Pharmacy,Harbin University of Commerce,Harbin 150076,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第8期30-36,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
黑龙江省博士后面上项目(LBH-Z19074)
哈尔滨市应用技术研究与开发项目(2017RAQXJ046)。
