海藻-甘草配伍对甲状腺肿大大鼠JNK1/Bcl-2/Beclin1通路表达的影响

doi:10.19803/j.1672-8629.20250710

摘要/Abstract

摘要： 目的观察海藻甘草（HG）反药组合应用对甲状腺肿大的治疗作用,探讨其作用与JNK1/Bcl-2/Beclin1通路调控的细胞自噬相关性。方法将大鼠随机分为空白（Control）、模型（Model）、优甲乐（Euthyrox）、海藻甘草高剂量（HG-H,7.92 g·kg^-1）、海藻甘草中剂量（HG-M,3.96 g·kg^-1）以及海藻甘草低剂量组（HG-L,1.98 g·kg^-1）6个组。除Control组外,其余组给予丙硫氧嘧啶（Propylthiouracil,PTU）灌胃14 d以复制甲状腺肿大大鼠病理模型,模型复制成功后各给药组给药14 d。在给药期间,为了稳定模型,每隔1 d给药组灌服PTU,剂量同前。基于大鼠甲状腺组织病理形态观察评估HG药效。通过免疫荧光、实时定量聚合酶链反应（Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction,RT-qPCR）和蛋白免疫印迹（Western Blot,WB）分析验证关键靶点。结果与Control组比较,Model组大鼠甲状腺系数升高（P<0.05）。与Model组相比,3个HG组大鼠甲状腺系数均下降（P<0.05）。在光镜下,与Control组相比,Model组滤泡形态不规则、甲状腺滤泡腔面积变小、滤泡上皮细胞高度增加和细胞核数量增加。与Model组相比,Euthyrox及各HG组甲状腺病理形态均有明显改善。LC3免疫荧光染色法结果显示,与Control组相比,Model组和HG-L组LC3荧光标记强度均增强（P<0.05）;与Model组相比,HG-L组LC3荧光强度降低（P<0.05）。通过RT-qPCR和WB检测,结果显示,与Control组相比,Model组大鼠甲状腺组织中JNK1、Beclin1、PI3KC3和LC3 mRNA及蛋白表达水平显著升高（P<0.05）,而Bcl-2mRNA及蛋白表达水平显著降低（P<0.05）。与Model组相比,HG-L组大鼠甲状腺组织中JNK1、Beclin1、PI3KC3和LC3mRNA及蛋白表达水平显著下调（P<0.05）,而Bcl-2mRNA及蛋白表达水平显著上调（P<0.05）。结论海藻甘草反药组合通过调控JNK1通路下游关键因子Bcl-2、Beclin1、PI3KC3和LC3表达水平,抑制甲状腺肿大细胞自噬进程,进而发挥其治疗作用。JNK1/Bcl-2/Beclin1信号通路可能是HG治疗甲状腺肿大关键分子机制。

关键词: 海藻, 甘草, 甲状腺肿大, 自噬, 作用机制, JNK1, Beclin1, 大鼠

Abstract: Objective To investigate the therapeutic effect of the incompatible Haizao-Gancao herbal pair against goitre, and to explore whether this effect is associated with autophagy regulated by the JNK1/Bcl-2/Beclin1 pathway. Methods Rats were randomly assigned to six groups: control, model, Euthyrox, high-dose Haizao-Gancao(HG-H, 7.92 g·kg^-1), medium-dose Haizao-Gancao (HG-M, 3.96 g·kg^-1), and low-dose Haizao-Gancao (HG-L, 1.98 g·kg^-1) groups. Except the control group, all the groups were orally administrated with propylthiouracil (PTU) for 14 days to replicate a pathological model of goiter rats. After modeling, each treatment group received their respective medication of 14 days. To stabilize the model, the treatment groups received oral PTU every other day at the previous dosage. The efficacy of HG was assessed based on histopathology of thyroid tissues of rats. Key targets were validated via immunofluorescence, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), and Western blot (WB) analysis. Results Compared with the control group, the thyroid coefficient increased in the model group (P<0.05). Compared with the model group, the thyroid coefficient decreased in all the three HG groups (P<0.05). Under light microscopy, irregular follicular morphology, smaller areas of thyroid follicular cavities, increased height of follicular epithelial cells, and larger numbers of nuclei were observed in the model group compared with the control group. Compared with the model group, the thyroid pathological morphology was significantly improved in the Euthyrox and all HG groups. LC3 immunofluorescence staining revealed enhanced LC3 fluorescence intensity in both the model and HG-L groups compared with the control group (P<0.05). However, the intensity of LC3 fluorescence decreased in the HG-L group relative to the model group (P<0.05). RT-qPCR and WB analysis suggested that mRNA and protein expression levels of JNK1, Beclin1, PI3KC3, and LC3 were significantly elevated in thyroid tissues of the model group (P<0.05) compared with the control group, while Bcl-2 mRNA and protein expression levels were significantly reduced (P<0.05). Compared with the model group, mRNA and protein expression levels of JNK1, Beclin1, PI3KC3, and LC3 were significantly downregulated in thyroid tissues in the HG-L group (P<0.05), while mRNA and protein expression levels of Bcl-2 were significantly upregulated (P< 0.05). Conclusion Haizao-Gancao herbal pair can inhibit cellular autophagy by regulating the expression levels of key downstream factors along the JNK1 pathway—Bcl-2, Beclin1, PI3KC3, and LC3, thereby exerting its therapeutic effect on goiter. The JNK1/Bcl-2/Beclin1 signaling pathway may be a key molecular mechanism underlying the therapeutic efficacy of Haizao and Gancao against goiter.

Key words: Haizao, Gancao, Goiter, Autophagy, Mechanism of Action, JNK1, Beclin1, Rat

中图分类号:

吴美晶, 徐向楠, 刘晓庆, 于雪, 钟赣生, 修琳琳. 海藻-甘草配伍对甲状腺肿大大鼠JNK1/Bcl-2/Beclin1通路表达的影响[J]. 中国药物警戒, 2026, 23(1): 55-62.

WU Meijing, XU Xiangnan, LIU Xiaoqing, YU Xue, ZHONG Gansheng, XIU Linlin. Effects of Haizao-Gancao Herbal Pair on JNK1/Bcl-2/Beclin1 Pathway Expressions in Rats with Goiter[J]. Chinese Journal of Pharmacovigilance, 2026, 23(1): 55-62.

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