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,长沙 401208;2. 湖南中医药大学第一附属医院,长沙 401007

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Establishment and evaluation of a rat model of perimenopausal dry eye with liver-kidney yin deficiency evidence
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1. Hunan University of Chinese Medicine, Changsha 401208, China; 2. the First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha 401007, China

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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10周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8.37 g/(kg·d)的剂量进行灌胃,假手术组和模型组大鼠按1 mL/100(g·d)灌胃生理盐水,持续21 d。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;酶联免疫吸附测定(ELISA)检测大鼠血清促卵泡生成素(FSH)、雌二醇(E2)和孕酮(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(IL-1β)、肿瘤坏死因子-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、体质量减轻、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-1β、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 establish 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 SPFgrade female SD rats were divided randomly into sham-operated, model, and Qiju Dihuang soup groups (n=10). Rats in the latter two groups underwent bilateral ovariectomy and were given 0.1% benzalkonium chloride eye drops combined with provocation for 10 weeks to establish a model of perimenopausal dry eye with liver and kidney yin deficiency. After modeling, rats in the Qiju Dihuang soup group were gavaged with Qiju Dihuang soup at a dose of 8.37 g/(kg·d), rats in the sham-operated and model groups were gavaged with saline at a rate of 1 mL/100 (g·d) for 21 d. The general condition, retinal screen test scores, tear secretion, time of tear film rupture, and corneal fluorescein staining were observed and recorded in each group. Serum levels of follicle-stimulating hormone (FSH), estradiol (E2), and progesterone (PROG) were measured by enzyme-linked immunosorbent assay. Pathological damage to the cornea and lacrimal glands were detected by hematoxylin/eosin staining. Expression levels of the inflammatory factors interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cornea were detected by immunohistochemistry. Results Compared with the sham-operated group, rats in the model group showed behavioral signs of slow action and reaction, irritability, body mass loss, and increased anal temperature (P<0.05), decreased retinal screen test scores (P<0.05), tear secretion (P<0.05), and time of tear film rupture (P<0.05), and pathological damage to the cornea and lacrimal glands. FSH levels increased and E2 and PROG levels decreased (P<0.05) and expression levels of IL-1 and TNF-α increased in the model group compared with the sham group. All the above indexes were significantly improved in the Qiju Dihuang soup group compared with the model group. Conclusions From the perspective of “measuring evidence by formula” in Chinese medicine, we successfully established a rat model of perimenopausal dry eye with liver-kidney yin deficiency syndrome, which provides a theoretical and experimental basis for future systematic and in-depth research on the mechanism of perimenopausal dry eye with formula and evidence.

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田赛男,刘桂成,陈瑶,刘培,彭俊,彭清华,胡淑娟.围绝经期干眼肝肾阴虚证大鼠模型的建立与评价[J].中国实验动物学报,2025,33(9):1270~1279.

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  • 收稿日期:2025-01-22
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  • 在线发布日期: 2025-10-21
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