围绝经期干眼肝肾阴虚证大鼠模型的建立与评价
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1.湖南中医药大学;2.湖南中医药大学第一附属医院;3.湖南中医药大学/湖南中医药大学第一附属医院

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国家自然科学基金项目(81574031);国家中医药管理局人才支持项目-岐黄学者(国中医药人教函[2022]6号);湖南省人才托举工程—年青优秀科技人才培养计划(2023TJ-N25);湖南省科技厅临床创新引导项目(2021SK51402);湖南省教育厅重点项目(22A0241,23A0300);湖南省中医药科研计划项目(A2023001);湖南省卫生健康委卫生科研课题一般课题(W20243198);湖南中医药大学校院联合基金重点项目(2023XYLHJJ005);湖南中医药大学校级科研项目一般项目(Z2023XJYB21);湖南中医药大学研究生创新课题项目(2024CX155).


Establishment and evaluation of a rat model of perimenopausal dry eye with liver and kidney yin deficiency evidence
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Affiliation:

1.Hunan University of Chinese Medicine;2.The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine;3.Hunan University of Chinese Medicine,The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine

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National Natural Science Foundation of China project (81574031); Talent Support Program of the National Administration of Traditional Chinese Medicine - Qihuang Scholar (Guo Zhong Yi Yao Ren Jiao Han [2022] No.6); Hunan Provincial Talent Support Program - Young and Excellent Scientific and Technological Talent Cultivation Plan (2023TJ-N25); Clinical Innovation Guidance Project of Hunan Provincial Science and Technology Department (2021SK51402); Key Project of Hunan Provincial Department of Education (22A0241, 23A0300); Hunan Provincial Traditional Chinese Medicine Scientific Research Program Project (A2023001); General Project of Health Research Topics from Hunan Provincial Health Commission (W20243198); Key Project of the Joint Fund of Hunan University of Chinese Medicine and its Affiliated Hospitals (2023XYLHJJ005); General Project of Hunan University of Chinese Medicine's School-level Scientific Research Program (Z2023XJYB21);Hunan University of Traditional Chinese Medicine Graduate Innovation Project(2024CX155).

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    摘要:

    目的 从“以方测证”的中医证候角度,探讨围绝经期干眼肝肾阴虚证大鼠模型的建立。方法 30只SPF级SD雌性大鼠随机分为假手术组、模型组、杞菊地黄汤组,每组10只。除假手术组外,其余2组大鼠行双侧卵巢切除,并予以0.1%苯扎氯铵滴眼液滴眼联合激怒法持续10w建立围绝经期干眼肝肾阴虚证模型。造模结束后,杞菊地黄汤组大鼠予以杞菊地黄汤按8.37g/(kg.d)的剂量进行灌胃,空白组和模型组大鼠按1ml/100(kg.d)灌入生理盐水,持续21d。观察并记录各组大鼠的一般情况、网屏实验评分、泪液分泌量、泪膜破裂时间、角膜荧光素染色;酶联免疫(enzyme-linkedimmunoassay, ELISA) 法检测大鼠血清促卵泡生成素(Follicle Stimulating Hormone, FSH)、雌二醇(Estradiol, E2)和孕酮(Progesterone, PROG)水平;苏木素-伊红(HE)染色观察大鼠角膜、泪腺病理损伤;免疫组化染色检测大鼠角膜中炎性因子白细胞介素-1β( interleukin-1, IL-1β)、肿瘤坏死因子α( tumor necrosis factor-α, TNF-α);结果 与假手术组对比,模型组大鼠在行为学上表现为行动迟缓、反应迟钝,易激怒,体重减轻,肛温上升(P<0.05),网屏实验评分下降(P<0.05),泪液分泌量减少(P<0.05),泪膜破裂时间缩短(P<0.05),角膜及泪腺病理损伤严重,血清性激素FSH水平升高,E2、PROG水平下降(P<0.05),泪腺炎性因子IL-1、TNF-α表达增加;与模型组比较,杞菊地黄汤组上述各指标均显著改善。结论 从“以方测证”的中医证候角度,成功建立了围绝经期干眼肝肾阴虚证病证结合大鼠模型,为今后围绝经期干眼方证对应机制的系统深入研究提供了理论和实验依据。

    Abstract:

    Objective To investigate the establishment of a rat model of perimenopausal dry eye with liver and kidney yin deficiency from the perspective of “measuring evidence by prescription” in Chinese medicine. Methods Thirty SPF-grade SD female rats were randomly divided into the sham-operated group, the model group, and the Qijvdihuang soup group, with 10 rats in each group. Except for the sham-operated group, the remaining 2 groups of rats underwent bilateral ovariectomy and were given 0.1% benzalkonium chloride eye drops combined with provocation method for 10 w to establish the model of perimenopausal dry eye with liver and kidney yin deficiency. At the end of modeling, rats in the Qijvdihuang soup group were gavaged with Qijvdihuang soup at a dose of 8.37 g/(kg·d), and rats in the blank group and the model group were gavaged with saline at a rate of 1 ml/100 (kg·d) for 21 d. The general conditions, the scores of retinal screen test, the amount of tear secretion, the time of tear film rupture, and the staining of corneal fluorescein of the rats in each group were observed and recorded; The enzyme-linked immunoassay (ELISA) method was used to measure serum Follicle Stimulating Hormone (FSH)、estradiol (E2) and progesterone (PROG) levels in rats; Hematoxycin-eosin (HE) staining was used to observe the pathological damage of cornea and lacrimal gland in rats; Immunohistochemical staining was used to detect the inflammatory factor interleukin-1β(IL-1β) and tumor necrosis factorα(TNF-α)in the cornea of rats

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  • 收稿日期:2025-01-22
  • 最后修改日期:2025-07-22
  • 录用日期:2025-09-05
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