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欧晨, 易肇兰, 宋厚盼, 谢薇, 彭清华.视网膜色素变性肝肾阴虚证小鼠的模型建立与评价[J].湖南中医药大学学报,2026,46(3):440-446[点击复制] |
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| 视网膜色素变性肝肾阴虚证小鼠的模型建立与评价 |
| 欧晨,易肇兰,宋厚盼,谢薇,彭清华 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208;湖南省妇幼保健院, 湖南 长沙 410008) |
| 摘要: |
| 目的 从“以方测证”的中医证候角度，探讨视网膜色素变性肝肾阴虚证小鼠的模型建立。方法 48只rd10小鼠采用情志刺激联合左甲状腺素钠片制备小鼠肝肾阴虚证候模型，采用随机数表法分为4组：模型组（等体积生理盐水）、乐盯组（乐盯软胶囊0.15 g/kg）、滋阴明目丸低剂量组（4.5 g/kg）、滋阴明目丸高剂量组（9 g/kg）；12只C57BL/6小鼠作为空白组（等体积生理盐水）。各组分别予以相应药物灌胃，每天1次，连续28 d。观察小鼠一般情况并记录小鼠头、舌、爪、尾的照片信息；ELISA检测小鼠血清环磷酸腺苷（cAMP）、环磷酸鸟苷（cGMP）、睾酮（T）含量；HE染色观察小鼠肝、肾、视网膜组织形态；视网膜血管网铺片过碘酸雪夫染色观察视网膜微血管形态。结果 与空白组比较，模型组小鼠精神萎靡，舌质红绛，少津，有裂纹，爪甲苍白，尾部皮肤干燥粗糙，可见鳞片状角化片；血清cAMP含量升高（P＜0.01），cGMP、T含量降低（P＜0.01）；肝细胞排列紊乱，出现萎缩及空泡变性，纤维组织增生；肾小管细胞排列紊乱，肾小球毛细血管壁缺损，肾组织内可见炎症细胞浸润及肾间质纤维化；视网膜明显萎缩变薄，视网膜色素上皮细胞缺失，外核层结构消失，视网膜厚度变薄（P＜0.01）；视网膜毛细血管排列紊乱，分布致密，管径变细，可见明显血管扭曲成节，相互攀缠。与模型组比较，乐盯组和滋阴明目丸低、高剂量组小鼠精神可，舌质红，爪甲色转红润，尾部皮肤鳞片状角化片减少或消失；血清cAMP含量显著降低（P＜0.01），cGMP、T含量升高（P＜0.05，P＜0.01）；肝、肾细胞排列较整齐，纤维组织增生减少；视网膜厚度增加（P＜0.01），视网膜毛细血管网分布较有序，管径增粗。与乐盯组比较，滋阴明目丸低剂量组血清cAMP含量升高（P＜0.01），滋阴明目丸低、高剂量组血清cGMP含量降低（P＜0.05，P＜0.01），视网膜厚度增加（P＜0.01）。结论 滋阴明目丸可以影响小鼠的一般状态，降低血清cAMP含量，升高cGMP和T含量，同时改善肝、肾、视网膜组织病理损伤，增加视网膜厚度。 |
| 关键词: 滋阴明目丸 视网膜色素变性 肝肾阴虚证 环磷酸腺苷 环磷酸鸟苷 睾酮 |
| DOI：10.3969/j.issn.1674-070X.2026.03.002 |
| 投稿时间：2025-10-12 |
| 基金项目:国家自然科学基金青年科学基金项目（82405490）；湖南省教育厅科学研究项目（23B0347）；中国博士后科学基金第76批面上资助项目（2024M760896）；湖南中医药大学校院联合基金项目（2024XYLH020）。 |
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| Establishment and evaluation of a mouse model of retinitis pigmentosa with liver-kidney yin deficiency pattern |
| OU Chen, YI Zhaolan, SONG Houpan, XIE Wei, PENG Qinghua |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Maternal and Child Health Care Hospital, Changsha, Hunan 410008, China) |
| Abstract: |
| Objective To establish a mouse model of retinitis pigmentosa (RP) with liver and kidney yin deficiency from the perspective of "determining pattern by formula" in Chinese medicine. Methods Forty-eight rd10 mice were used to establish a liver-kidney yin deficiency pattern model through emotional stimulation combined with levothyroxine sodium tablet. The mice were randomly divided into four groups using random number table method: model group (equal volume of normal saline), Leding group (0.15 g/kg of Leding soft capsules), low-dose Ziyin Mingmu Pill (ZYMMP) group (4.5 g/kg), and high-dose ZYMMP group (9 g/kg). Twelve C57BL/6 mice served as the blank group (equivalent volume of normal saline). Each group was given the corresponding drug by intragastric administration once a day for 28 consecutive days. General conditions of the mice were observed, and photographic information of their heads, tongues, claws, and tails were recorded. Serum levels of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate(cGMP) and mouse testosterone (T) were measured using ELISA. The morphological changes in the liver, kidney, and retinal tissues of mice were observed via HE staining, and retinal microvascular morphology was examined using vascular network flattening and periodic acid-Schiff (PAS) staining. Results Compared with the blank group, mice in the model group showed signs of lethargy, dark red and dry tongues with cracks, pale claws and nails, as well as dry and rough tail skin with scaly keratinized patches. Serum cAMP levels increased (P<0.01), while cGMP and T levels decreased (P<0.01). Hepatocytes were disorganized with visible atrophy, vacuolar degeneration, and fibrous tissue hyperplasia. Renal tubular cells were irregularly arranged, glomerular capillary walls were defective, and inflammatory cell infiltration and renal interstitial fibrosis were observed. The retina was significantly atrophied and thinner, with loss of retinal pigment epithelial cells and disappearance of the outer nuclear layer structure. Retinal thickness decreased (P<0.01). Retinal capillaries were disorganized, densely distributed, and thinner in diameter, with noticeable vascular twisting and entanglement. Compared with the model group, Leding group, low-dose and high-dose ZYMMP group showed normal mental state, with red tongue, reddish and moist claws and nails, and a reduction or disappearance of scaly keratinized patches on the tail. cAMP levels significantly decreased (P<0.01), while cGMP and T levels increased (P<0.05 or P<0.01). Liver and kidney cells were more regularly arranged, with reduced fibrous tissue hyperplasia. Retinal thickness significantly increased (P<0.01). The retinal capillary network was more orderly distributed, with thickened vessel diameters. Compared with the Leding group, the low-dose ZYMMP group showed an increase in cAMP levels (P<0.01), while both the low- and high-dose ZYMMP groups exhibited a decrease in cGMP levels (P<0.05, P<0.01) and an increase in retinal thickness (P<0.01). Conclusion ZYMMP can improve the general state of mice, reduce serum cAMP levels, elevate cGMP and T levels, and concurrently ameliorate pathological damage in the liver, kidney, and retina, as well as increases retinal thickness. |
| Key words: Ziyin Mingmu Pill retinitis pigmentosa liver kidney yin deficiency pattern cyclic adenosine monophosphate cyclic guanosine monophosphate testosterone |
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