围绝经期综合征肾阴虚证大鼠汗液分泌及其生化指标的变化
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福建中医药大学 药学院

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国家中医药管理局临床中药学高水平中医药重点学科建设项目(国中医药人教函[2022]226号),福建省自然科学基金资助项目(2023J01857),福建中医药大学基础类学科科研提升项目(XJC2022013)


Study on the sweat secretion and the biochemical indexes in Yin-deficiency ovariectomized rats
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College of Pharmacy, Fujian University of Traditional Chinese Medicine

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High level Key Discipline Construction Project of Clinical Traditional Chinese Medicine of the State Administration of Traditional Chinese Medicine (Guo Zhong Yao Ren Jiao Han [2022] No. 226), Fujian Natural Science Foundation funded project (2023J01857), and Fujian University of Traditional Chinese Medicine Basic Discipline Research Enhancement Project (XJC2022013)

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    摘要:

    目的 评估围绝经期综合征肾阴虚证“病证结合”模型大鼠的汗液分泌情况,并探讨其生化指标的变化。方法 将18只SD雌性大鼠随机分为空白对照组、模型组、阳性药组,每组6只。空白对照组行假手术,模型组和阳性药组大鼠行双侧去卵巢手术,术后第7天给予L-甲状腺素(92 mg·kg-1)每日1次,连续给药7天建立围绝经期综合征肾阴虚证“病证结合”模型。阳性药组每日灌胃清骨散汤剂(7.3 g·kg-1),假手术组和模型组灌胃等量蒸馏水,每日1次,共14天。采用和田-高垣试剂着色法测定大鼠足跖部汗液分泌。酶联免疫吸附试验(ELISA)检测大鼠血清cAMP、cGMP、LH、 GnRH、E2水平。蛋白免疫印迹法(Western blot)法测定大鼠足趾肉垫组织的M3受体、β2受体、水通道蛋白5(Aquaporin,AQP5)表达水平。 结果 (1)与假手术组比,模型大鼠体温升高,双耳发红,易激惹、毛发干枯等阴虚证的典型行为学表现,模型组大鼠汗液分泌显著增多;(2)模型大鼠血清cAMP水平显著升高, cGMP水平显著降低,cAMP/cGMP比值显著升高,LH、GnRH水平显著升高,E2水平显著降低。(3)大鼠足趾组织的M3受体表达下调,β2受体、AQP5表达水平上调。阳性药治疗2周后,血清cAMP水平显著下降, cAMP/cGMP比值显著降低,LH、GnRH水平降低,但不影响血清E2水平。大鼠足趾组织的M3受体上调,β2受体、AQP5表达水平下调。 结论 去势联合甲状腺素建立的围绝经期综合征肾阴虚证“病证结合”模型大鼠汗液分泌显著增多,其机制可能与cGMP、cAMP及其在汗腺上调控汗液分泌的关键蛋白M3R,β2AR,AQP5的变化有关。

    Abstract:

    Objective To assess the sweat secretion of Yin-deficiency ovariectomized rats and to investigate the changes of biochemical indexes. Methods Eighteen SD female rats were randomly divided into sham operation group, model group, and positive control group with six rats in each group. The sham operation group were taken sham operation, the model group and the positive control group underwent bilateral ovary removal surgery, and L-thyroxine (92mg·kg-1) was given once a day from the 7th day after the operation for 7 consecutive days to establish Yin-deficiency ovariectomized model. The positive control group was orally administered Qinggu San Tang (7.3g·kg-1) daily, while the sham operation group and model group were orally administered an equal amount of distilled water once a day for a total of 14 days. Sweat secretion from the rat foot-plantar region was determined using the Wada-Takagaki reagent colouring method. At the end of the experiment,blood was taken from the abdominal aorta, and the tissue of the paw pads of rats were separated. Serum cAMP, cGMP, LH, GnRH, and E2 levels were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to determine the expression levels of M3 receptor, β2 receptor, and aquaporin-5(AQP5) in the paw pad. Results (1) Compared with the sham operation group, rats in the model group were more irritable and aggressive, their body weights decreased while their average temperature increased significantly, sweat secretion increased significantly; (2) Serum cAMP and cAMP/cGMP ratio increased, LH and GnRH increased significantly, and E2 level decreased; (3) M3 receptor expression was down-regulated and β2 receptor, aquaporin (AQP5) expression up-regulated in paw pads of rats. After 2 weeks of positive control treatment, serum cAMP and the ratio of cAMP/cGMP decreased significantly, LH and GnRH levels decreased, but no statistical difference was observed for serum E2 level. The expression levels of M3 receptor, β2 receptor, and aquaporin (AQP5) were down-regulated in paw pads of rats. Conclusion: Sweat secretion was significantly increased in the perimenopausal syndrome kidney yin deficiency "combined disease and evidence" model established by desiccation combined with thyroxine, and the mechanism may be related to the changes of cGMP, cAMP, and its key proteins M3R, β2AR, and AQP5 in sweat glands that regulate sweat secretion.

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  • 收稿日期:2023-11-29
  • 最后修改日期:2024-04-01
  • 录用日期:2024-05-17
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