目的：探讨富组氨酸糖蛋白(HRG)在大鼠糖尿病视网膜病变新生血管形成中的作用。

方法：建立链脲佐菌素(STZ)诱导的SD大鼠糖尿病模型，蛋白免疫印迹法(WB)检测正常(WT)组 、糖尿病(DM)组视网膜中HRG、血管生成因子(VEGF)的蛋白表达情况。转染HRG小干扰RNA低表达序列，WB验证在高糖诱导的人视网膜微血管内皮细胞(hRMECs)中HRG的蛋白表达情况，选择最佳si-HRG#298序列用于后续实验。动物实验通过腺相关病毒载体沉默HRG，玻璃体腔注射HRG空载体对照组(AAV2-sh-NC)及HRG基因沉默组(AAV2-sh-HRG#298)，WB验证HRG的蛋白表达情况后，通过HE染色观察各组视网膜结构变化，PAS染色观察各组视网膜新生血管变化情况，WB检测各组HRG及VEGF的蛋白表达情况。

结果：HE染色发现DM组大鼠视网膜结构出现紊乱，神经节细胞层细胞数量减少，内核层和外核层细胞数量减少，视网膜总厚度也减少(P<0.05)； PAS染色观察DM组大鼠视网膜中无细胞毛细血管明显增多(P<0.05)； DM组大鼠视网膜中HRG及血管生成因子VEGF的蛋白表达上调(P<0.05)； 高糖诱导的hRMECs中转染HRG后其蛋白表达明显下调(P<0.05)； HRG基因沉默可以抑制糖尿病视网膜的新生血管形成，下调VEGF的蛋白表达(P<0.05)。

结论：HRG促进糖尿病大鼠视网膜的新生血管形成，HRG基因沉默可以抑制其新生血管形成。

METHODS: Streptozocin(STZ)-induced diabetic Sprague-Dawley(SD)rats were utilized as an experimental model, the protein expression of HRG and vascular endothelial growth factor(VEGF)in the retinas of normal(Wild type, WT)and diabetic(diabetic mellitus, DM)groups was detected using Western blot(WB). The protein expression of HRG in high-glucose-induced human retinal microvascular endothelial cells(hRMECs)was verified by WB after transfection with HRG small interfering RNA(siRNA)low-expression sequences. The optimal si-HRG#298 sequence was selected for further experiments. In the animal experiment, HRG gene silencing was achieved using an adeno-associated virus(AAV)vector, with AAV2-sh-NC and AAV2-sh-HRG#298 serving as the HRG gene silencing group and the HRG empty vector control group, respectively. The protein expression of HRG and VEGF in each group was then detected by WB following the verification of HRG protein expression. Retinal structural changes were observed by HE staining, and neovascularization changes were observed by PAS staining.

RESULTS: HE staining found that the retinal structure in the DM group was disordered, the number of cells in the ganglion cell layer decreased, the number of cells in the inner and outer nuclear layers decreased, and the total retinal thickness also decreased(P<0.05); cellular capillaries were significantly increased in DM rats observed by PAS staining(P<0.05); the protein expression of HRG and angiogenesis factor VEGF was up-regulated in the retina of DM group(P<0.05); the protein expression of HRG was significantly downregulated in high glucose-induced hRMECs(P<0.05); the inhibition of neovascularization in diabetic retinas and the downregulation of VEGF protein expression were achieved through HRG gene silencing(P<0.05).

CONCLUSION: HRG promotes neovascularization in the retinas of diabetic rats, and HRG gene silencing can inhibit neovascularization.