摘要
目的探讨海藻玉壶汤对甲亢大鼠Bax、Bcl-2 mRNA表达的影响。方法选取SPF级SD大鼠60只,雌雄各半,随机分为空白组、模型组,其中空白组10只,模型50只;模型组大鼠皮下注射L-左旋甲状腺素钠造模,1次/d,造模时间为1周,单次给药剂量0.35 mg/kg,造模结束后将大鼠分为模型组、甲巯咪唑(MMI)组,海藻玉壶汤低、中、高剂量组。给药组灌胃时间为8周,空白组与模型组给予等量的0.9%生理盐水灌胃,中药高、中、低各组剂量为42.86、21.43、12.44 g/kg,甲巯咪唑(MMI)组剂量为0.1875 g/kg。灌胃第0、8周应用酶联免疫吸附(ELISA)法检测各组大鼠T3、T4、TSH,灌胃结束后应用实时荧光定量PCR法(qPCR)检测各组甲状腺组织中Bax、Bcl-2 mRNA的表达,苏木素-伊红(HE)染色观察各组大鼠甲状腺组织病理形态,免疫荧光法检测大鼠甲状腺Bax、Bcl-2的表达。结果与空白组比较,模型组T3、T4显著升高,TSH下降(P<0.05);灌胃结束后,与模型组相比,甲巯咪唑(MMI)组、中药中高剂量组T3、T4下降,TSH上升(P<0.05),中药低剂量组T3、T4、TSH无明显改变。病理切片显示,模型组与空白组比较,甲状腺部分区域内甲状腺滤泡上皮增生,滤泡增大,大小形态不一。甲巯咪唑(MMI)组、中药中高剂量组甲状腺组织与模型组比较,甲状腺滤泡上皮无明显增生,滤泡大小形态不一,滤泡肿大较轻。免疫荧光结果显示:模型组与空白组比较,甲状腺中Bcl-2表达量增加,与模型组比较,甲巯咪唑(MMI)组、中药中高剂量组甲状腺中Bcl-2表达量下降,所有组甲状腺中Bax无明显变化。实时荧光定量聚合酶链式反应(qPCR)结果显示:与空白组比较,模型组甲状腺中Bcl-2 mRNA表达量明显增加(P<0.05);与模型组比较,甲巯咪唑(MMI)组、中药中高剂量组甲状腺中Bcl-2 mRNA表达量下降(P<0.05);各组甲状腺中Bax mRNA表达量无明显差异(P>0.05)。结论海藻玉壶汤可通过抑制大鼠甲状腺Bcl-2 mRNA的表达改善甲亢大鼠病情,但对Bax mRNA的表达无明显影响。
Objective To study the effect of Haizao Yuhu Tang on the expression of Bax and Bcl-2 mRNA in hyperthyroidism rats.Methods 60 SPF grade SD rats,half male and half female,were randomly divided into a blank group of 10 rats and a model group of 50 rats.The rats in model group were subcutaneously injected with L-neneneba levothyroxine sodium once a day for one week,with a single dose of 0.35 mg/kg.After the completion of modeling,they were divided into model group,western medicine(MMI)group,low,medium and high dose groups of Chinese medicine(Haizao Yuhu Decoction).The drug group was given medication by gavage for 8 weeks.The blank group and model group were given an equal amount of 0.9%physiological saline by gavage.The high,medium and low dose groups of traditional Chinese medicine received doses of 42.86 g/kg,21.43 g/kg,12.44 g/kg,and the western medicine(MMI)group received doses of 0.1875 g/kg.Enzyme linked immun-osorbent assay(ELISA)was used to detect T3,T4 and TSH in each group of rats at week 0 and 8 during the gavage period.After the gavage,real-time fluorescence quantitative PCR(qPCR)was used to detect the expression of Bax and Bcl-2 mRNA in the thyroid tissue of each group.Hematoxylin eosin(HE)staining was used to observe the pathological morphology of the thyroid tissue of rats in each group.Result Compared with the blank group,the model group showed a significant increase in T3 and T4,while TSH decreased(P<0.05);After gavage,compared with the model group,the western medicine MMI group,the middle and high dose groups of Haizao Yuhu Tang showed a decrease in T3 and T4,while the high dose group showed an increase in TSH(P<0.05).The low dose group of Haizao Yuhu Tang showed no significant changes in T3,T4 and TSH.Pathological sections show that compared with the blank group,the thyroid tissue in the model group shows epithelial hyperplasia of thyroid follicles in some areas,with enlarged follicles and varying sizes and shapes.Compared with the model group,the high and medium dose groups of Haizao Yuhu Tang and MMI group showed no significant proliferation of thyroid follicle epithelium,varying sizes and shapes of follicles,and milder follicular enlargement.The immunofluorescence results showed that compared with the blank group,the expression of Bcl-2 in the thyroid gland of the model group increased.Compared with the model group,the expression of Bcl-2 in the thyroid gland of the high and medium dose groups of Haizao Yuhu Tang and MMI group decreased.The expression of Bcl-2 in the thyroid gland of the high and medium-dose group of Haizao Yuhu Tang and MMI decreased,and there was no significant change in Bax in the thyroid gland of all groups.Real time fluorescence quantitative Polymerase chain reaction(qPCR)results showed that the expression level of Bcl-2 mRNA in the thyroid gland of the model group was significantly increased(P<0.05);Compared with the model group,the expression of Bcl-2 mRNA in the thyroid gland of the western medicine MMI group,the Haizao Yuhu Tang medium and high-dose groups decreased(P<0.05);There was no significant difference in the expression of Bax mRNA in the thyroid of each group.Conclusion Haizao Yuhu Tang can improve the condition of hyperthyroidism in rats by inhibiting the expression of Bcl-2 mRNA in the thyroid gland of rats,but has no obvious effect on the expression of Bax mRNA.
作者
苏炳华
蔡萧君
陈星海
李树延
杨羽飞
SU Binghua;CAI Xiaojun;CHEN Xinghai;LI Shuyan;YANG Yufei(Traditional Chinese Medicine Academy of Heilongjiang,Harbin 150036,China)
出处
《北华大学学报(自然科学版)》
CAS
2024年第2期178-184,共7页
Journal of Beihua University(Natural Science)
基金
黑龙江省卫生健康委科技计划项目(20212121020028)。