目的 探讨荆防颗粒对db/db小鼠血糖代谢的调控作用及对其视网膜病变（diabetic retinopaphy，DR）的保护作用。方法 将60只雄性db/db小鼠随机分为模型组、二甲双胍（120 mg/kg）组及荆防颗粒（1、2 g/kg）组，每组15只，另取15只雄性db/m小鼠作为对照组，每周检测小鼠体质量、空腹血糖、摄食量及饮水量等基础指标，并在实验结束后进行口服葡萄糖耐量试验（oral glucose tolerance test，OGTT）；采用ELISA检测小鼠视网膜组织白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、丙二醛（malondialdehyde，MDA）水平和过氧化氢酶（catalase，CAT）、超氧化物歧化酶（superoxide dismutase，SOD）活性；采用PAS染色检测视网膜组织病理变化；采用Western blotting检测视网膜组织中血管内皮生长因子（vascular endothelial growth factor，VEGF）、紧密连接跨膜蛋白（Claudin-1）、胰岛素受体底物1（insulin receptor substrate 1，IRS1）、磷酸化IRS1（phosphorylated IRS1，p-IRS1）、磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）、磷酸化PI3K（phosphorylated PI3K，p-PI3K）、蛋白激酶B（protein kinase B，Akt）、磷酸化Akt（phosphorylated Akt，p-Akt）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）蛋白表达。结果 与对照组比较，模型组小鼠空腹血糖、摄食量、饮水量明显升高（P＜0.01），OGTT中曲线下面积（AUC）显著升高（P＜0.01）；视网膜组织中IL-1β、IL-6、TNF-α和MDA含量显著升高（P＜0.01），CAT、SOD活性显著下降（P＜0.01）；视网膜组织周细胞降低，新生血管增多（P＜0.01），并且视网膜组织中Claudin-1、p-IRS1/IRS1、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白表达下调（P＜0.01），VEGF、Bax、cleaved Caspase-3蛋白表达上调（P＜0.01）。与模型组比较，荆防颗粒组小鼠体质量明显升高（P＜0.05、0.01），空腹血糖、摄食量、饮水量均有不同程度下降（P＜0.05、0.01）；视网膜组织中IL-1β、IL-6、TNF-α、MDA含量显著下降（P＜0.05、0.01），CAT、SOD活性显著上升（P＜0.05、0.01）；视网膜组织病理形态有所改善，视网膜组织中Claudin-1、p-IRS1/IRS1、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白表达显著上调（P＜0.05、0.01），VEGF、Bax、cleaved Caspase-3蛋白表达显著下调（P＜0.05、0.01）。结论 荆防颗粒对db/db小鼠视网膜病变具有一定的改善作用，其作用机制可能为调节小鼠视网膜组织中VEGF及Claudin-1蛋白表达，进而激活胰岛素受体IRS-1/PI3K/Akt信号通路，启动Caspase级联反应，抑制视网膜上皮细胞的凋亡，从而对视网膜病变起到一定的保护作用。

Objective To investigate the regulation of Jingfang Granules (荆防颗粒) on blood glucose metabolism and protective effect of diabetic retinopaphy (DR) in db/db mice. Methods A total of 60 male db/db mice were randomly divided into model group, metformin (120 mg/kg) group, Jingfang Granules (10, 20 mg/kg) groups, with 15 mice in each group, and 15 male db/m mice were selected as control group. The basic indexes such as body weight, fasting blood glucose, food intake and water intake of mice were detected weekly. After the experiment, oral glucose tolerance test (OGTT) was detected; The levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), malondialdehyde (MDA) and the activities of catalase (CAT) and superoxide dismutase (SOD) in mouse retinal tissue were detected by ELISA; PAS staining was used to detect the pathological changes of retinal tissue; Western blotting was used to detect the protein expressions of vascular endothelial growth factor (VEGF), tight junction transmembrane protein (Claudin-1), insulin receptor substrate 1 (IRS1), phosphorylated IRS1 (p-IRS1), phosphatidylinositol 3-kinase (PI3K), phosphorylated PI3K (p-PI3K), protein kinase B (Akt), phosphorylated Akt (p-Akt), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), and cleaved cystein-asparate protease-3 (cleaved Caspase-3) in retinal tissue. Results Compared with control group, fasting blood glucose, food intake and water intake of mice in model group were significantly increased (P < 0.01), AUC in OGTT was significantly increased (P < 0.01); The levels of IL-1β, IL-6, TNF-α, MDA in retinal tissue were increased (P < 0.01). CAT and SOD activities were decreased (P < 0.01); The number of peripheral cells in retinal tissue was decreased and the number of new blood vessels was increased (P < 0.01), protein expressions of Claudin-1, p-IRS1/IRS1, p-PI3K/PI3K, p-Akt/Akt and Bcl-2 were down-regulated (P < 0.01), VEGF, Bax and cleaved Caspase-3 protein expressions were up-regulated (P < 0.01). Compared with model group, body weight of mice in Jingfang Granules group was significantly increased (P < 0.05, 0.01), fasting blood glucose, food intake and water intake were decreased to varying degrees (P < 0.05, 0.01); The levels of IL-1β, IL-6, TNF-α and MDA in retinal tissue were decreased (P < 0.05, 0.01). CAT and SOD activities were increased (P < 0.05, 0.01); The histopathologic morphology of retina was improved, and the protein expressions of Claudin-1, p-IRS1/IRS1, p-PI3K/PI3K, p-Akt/Akt, Bcl-2 in retinal tissue were up-regulated (P < 0.05, 0.01), VEGF, Bax and cleaved Caspase-3 protein expressions were down-regulated (P < 0.05, 0.01). Conclusion Jingfang Granules can improve retinopathy in db/db mice to a certain extent, and its mechanism may be related to regulating the expressions of VEGF and Claudin-1 protein in retinal tissue of mice, thus activating the insulin receptor IRS-1/PI3K/Akt signaling pathway, initiating the Caspase cascade reaction, and inhibiting the apoptosis of retinal epithelial cells. Thus, it has a certain protective effect on retinopathy.

R285.5

山东省重大科技创新工程（2020CXGC010505，2021CXG010508）