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胡卓瑜,王萱,胡齐,刘静,黄栊瑢,付美林,刘志敏,陈向东.灯盏花素对大鼠糖尿病视网膜水肿PKCβ及VEGF蛋白表达的影响[J].湖南中医药大学学报,2023,43(10):1762-1770[点击复制] |
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| 灯盏花素对大鼠糖尿病视网膜水肿PKCβ及VEGF蛋白表达的影响 |
| 胡卓瑜,王萱,胡齐,刘静,黄栊瑢,付美林,刘志敏,陈向东 |
| (湖南中医药大学, 湖南 长沙 410208;湖南中医药大学第一附属医院, 湖南 长沙 410007;中医药防治眼耳鼻喉疾病湖南省重点实验室, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨灯盏花素对链脲霉素（streptozotocin, STZ）诱导视网膜水肿大鼠血-视网膜屏障的作用及对蛋白激酶C β(protein kinases C β, PKCβ)及血管内皮生长因子（vascular endothelial growth factor, VEGF)蛋白表达的影响。方法 除正常组正常饲养外，其他各组大鼠用30 mg/kg剂量一次性腹腔注射STZ联合高脂高糖饲料饮食诱导SD大鼠糖尿病模型。HE染色及OCT观察视网膜组织形态，确立成功建立糖尿病视网膜水肿模型。造模成功后，随机分为模型组、安多明组及灯盏花素组，予以用药4周，观察大鼠的血糖、体质量变化，行OCT检查。通过HE染色观察视网膜组织形态组织学变化，Western blot检测视网膜组织VEGF、PKCβ蛋白的表达水平，ELISA法检测血清VEGF、PKCβ的表达水平，免疫组织化学检测视网膜Claudin-5、Occludin的表达水平。结果 与空白组对比，造模后2、4、8周以及给药后1、2、4周，模型组、安多明组、灯盏花素组血糖显著升高（P＜0.01）；且体质量增长缓慢（P＜0.01），并有体质量下降趋势；视网膜神经节细胞呈空泡样改变，内丛状层、外丛状层排列紊乱，内核层、外核层细胞密度减少；视网膜厚度增加（P＜0.01）；视网膜组织及血清中VEGF、PKCβ水平均显著上升（P＜0.01）；视网膜组织claudin-5、occludin的表达量下降（P＜0.01）。给药4周后,与模型组对比，灯盏花素组层间水肿不明显，视网膜厚度减少（P＜0.01）；视网膜组织PKCβ、VEGF水平显著降低（P＜0.01）；血清PKCβ、VEGF水平显著降低（P＜0.01）；视网膜组织claudin-5、occludin的表达量上升（P＜0.05，P＜0.01）。结论 灯盏花素可以下调VEGF、PKCβ的表达水平，使得claudin-5、occludin的表达量升高，以修复STZ诱导糖尿病视网膜水肿大鼠的血-视网膜屏障损伤。 |
| 关键词: 糖尿病视网膜水肿 灯盏花素 蛋白激酶Cβ 血管内皮生长因子 链脲霉素 血-视网膜屏障 |
| DOI：10.3969/j.issn.1674-070X.2023.10.003 |
| 投稿时间：2023-03-23 |
| 基金项目:湖南省自然科学基金项目（2021JJ30520）；湖南省中医药科研计划项目重点课题（C2022003）；湖南省中医药科研计划项目一般指导课题（E2022044）；湖南中医药大学研究生创新课题（2023CX09）；湖南省教育厅科学研究重点项目（22A0250）；湖南省中医药管理局重点项目（C2023013）；湖南中医药大学研究生创新课题（2022CX150）。 |
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| Effects of scutellarin on protein expressions of PKCβ and VEGF in rats with diabetic retinal edema |
| HU Zhuoyu,WANG Xuan,HU Qi,LIU Jing,HUANG Longrong,FU Meilin,LIU Zhimin,CHEN Xiangdong |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan Key Laboratory for Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the effects of scutellarin (Scu) on the blood-retinal barrier and the expressions of protein kinase C β (PKCβ) and vascular endothelial growth factor (VEGF) protein in rats with streptozotocin (STZ)-induced retinal edema. Methods In addition to normal feeding group, the SD rats in other groups were induced into diabetes model by intraperitoneal injection of STZ at a dose of 30 mg/kg in combination with a high-fat and high-sugar diet. HE staining and OCT were used to observe the retinal tissue morphology and a model of diabetes retinal edema was successfully established. After successful modeling, the rats were randomly divided into model group, andomin group, and scutellarin group. They were administered medications for 4 weeks to observe the changes in blood glucose and body weight, and were examined by OCT. In addition, HE staining was used to observe the retina. The protein expressions of VEGF and PKCβ in retinal tissue were determined by Western blot, and expression levels of VEGF and PKCβ in serum were examined by ELISA respectively. Immunohistochemistry was used to measure the expression levels of claudin-5 and occludin in the retina. Results Compared with the blank group, blood glucose levels significantly increased in model group, andomin group, and scutellarin group at 2, 4, and 8 weeks after modeling, as well as at 1, 2, and 4 weeks after administration (P＜0.01). The body weight showed a slow growth (P＜0.01) with a tendency to decrease; the retinal ganglion cells showed vacuolar changes, the inner plexiform layer and outer plexiform layer were in disorder, and the cell density of both inner and outer nuclear layers decreased; retinal thickness increased (P＜0.01). The levels of VEGF and PKCβ in retinal tissue and serum significantly increased (P＜0.01); the expression levels of claudin-5 and occludin in retinal tissue decreased (P＜0.01). After 4 weeks of administration, compared with the model group, the interlayer edema was not significant, the retinal thickness decreased (P＜0.01), the levels of PKCβ and VEGF in retinal tissue decreased (P＜0.01), the levels of PKCβ and VEGF in serum also significantly decreased (P＜0.01), while the expression levels of claudin-5 and occludin in retinal tissue increased (P＜0.05, P＜0.01) in scutellarin group. Conclusion Scutellarin can down-regulate the expression levels of VEGF and PKCβ, increase the expressions of claudin-5 and occludin, so as to repair the blood-retinal barrier damage in STZ-induced diabetic retinal edema rats. |
| Key words: diabetes retinal edema scutellarin protein kinase C β vascular endothelial growth factor streptozotocin blood-retinal barrier |
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