摘要
【目的】观察糖眼宁对糖尿病大鼠视网膜的保护作用。【方法】采用腹腔注射链脲佐菌素建立糖尿病大鼠模型。将造模成功的大鼠随机分为模型组,导升明组及糖眼宁高、中、低剂量组,每组10只。另设正常组。糖眼宁高、中、低剂量组大鼠分别给予糖眼宁10.8、5.4、2.7 g·kg^(-1)·d^(-1)灌胃,导升明组大鼠给予导升明0.135 g·kg^(-1)·d^(-1)灌胃,正常组、模型组给予等体积蒸馏水灌胃。灌胃期间每4周检测大鼠体质量和空腹血糖。灌胃12周后,分别采用苏木素-伊红染色、末端脱氧核苷酸转移酶介导的d UTP缺口末端标记(TUNEL)染色法观察大鼠视网膜组织病理形态,采用免疫组织化学法检测视网膜组织凋亡相关斑点样蛋白(ASC)表达,免疫印迹分析(Western Blot)法检测视网膜组织焦亡相关蛋白BRCC36、Nod样受体蛋白3(NLRP3)、ASC、半胱氨酸天冬氨酸蛋白酶1前体(Pro-Caspase-1)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、消皮素D(GSDMD)的表达情况。【结果】(1)给药4周后,与正常组比较,模型组体质量趋于降低,差异有统计学意义(P<0.05);给药4周后,与模型组比较,导升明组和糖眼宁低、中剂量组体质量变化差异无统计学意义(P>0.05),糖眼宁高剂量组体质量持续增加,差异有统计学意义(P<0.05)。(2)与正常组比较,模型组大鼠空腹血糖水平明显升高(P<0.05);与模型组比较,导升明组和糖眼宁各剂量组大鼠空腹血糖水平无明显降低,差异无统计学意义(P>0.05)。(3)与正常组比较,模型组大鼠视网膜萎缩,层间细胞排列紊乱;与模型组比较,导升明组和糖眼宁各剂量组大鼠视网膜组织结构改善。(4)与正常组比较,模型组大鼠视网膜凋亡细胞染色程度、ASC表达明显增强(P<0.05);与模型组比较,导升明组和糖眼宁各剂量组大鼠视网膜凋亡细胞染色程度、ASC表达不同程度减弱(P<0.05)。(5)与正常组比较,模型组大鼠焦亡关键蛋白NLRP3、ASC、Pro-Caspase-1、Caspase-1、GSDMD、BRCC36表达水平明显升高(P<0.05);与模型组比较,导升明组和糖眼宁各剂量组视网膜组织NLRP3、ASC、Pro-Caspase-1、Caspase-1、GSDMD、BRCC36表达水平明显降低(P<0.05)。【结论】糖眼宁能够减轻糖尿病大鼠视网膜炎症反应,抑制视网膜细胞焦亡,进而维护血-视网膜屏障的完整性。
Objective To observe the protect effect of Tangyanning on retina in diabetic rats.Methods The diabetic rat model was established by intraperitoneal injection of streptozotocin.The successfully modeled rats were randomly divided into model group,Doxium group and Tangyanning high-dose,medium-dose and low-dose groups,with 10 rats in each group.Another normal group was set up.Rats in Tangyanning high-,medium-and low-dose groups were given intragastric administration of 10.8,5.4,2.7 g·kg^(-1)·d^(-1) of Tangyanning,respectively,rats in Daoshengming group were given intragastric administration of 0.135 g·kg^(-1)·d^(-1) of Daoshengming,normal group and model group were given equal volume of distilled water intragastric administration.Body mass and fasting blood glucose level were measured every 4 weeks during gavage.After 12 weeks of intragastric administration,the histopathological morphology of retina was observed by hematoxylin-eosin staining and terminal dUTP nick end labeling(TUNEL)staining,and the expression of apoptosis-related spotted protein(ASC)in retinal tissue was detected by immunohistochemistry.Western Blot was used to detect the expression levels of pyrotopic-associated proteins BRCC36,NLR family pyrin domain containing 3(NLRP3),ASC,Pro-Caspase-1,Caspase-1 and gasdermin D(GSDMD)in retinal tissues.Results(1)After 4 weeks of medication,compared with the normal group,body mass in the model group tended to decrease(P<0.05);after 4 weeks of medication,compared with the model group,there was no significant decrease in body mass in the Daoshengming group and Tangyanning medium-dose and low-dose groups,the difference being statistically significant(P>0.05),while body mass in the Tangyanning high-dose group tended to increase(P<0.05).(2)Compared with the normal group,fasting blood glucose level in the model group was significantly increased(P<0.05);compared with the model group,there was no significant decrease in fasting blood glucose level in the Daoshengming and Tangyanning dose groups,the difference being statistically significant(P>0.05).(3)Compared with the normal group,retinal atrophy and disorder of interlamellar cells were observed in the model group;compared with the model group,the retinal tissue structure of the Daoshengming and Tangyanning dose groups was improved.(4)Compared with the normal group,the staining degree of apoptotic cells and ASC expression in the model group were significantly increased(P<0.05);compared with model group,the staining degree of retinal apoptotic cells and ASC expression in the Daoshengming and Tangyanning dose groups were decreased to different degrees(P<0.05).(5)Compared with the normal group,the expression levels of NLRP3,ASC,Pro-Caspase-1,Caspase-1,GSDMD and BRCC36 in the model group were significantly increased(P<0.05);Compared with the model group,the expression levels of NLRP3,ASC,Pro-Caspase-1,Caspase-1,GSDMD and BRCC36 in the Daoshengming group and Tangyanning dose groups were significantly decreased(P<0.05).Conclusion Tangyanning exerts effect in reducing the inflammatory response of the retina in diabetic rats,inhibiting the pyroptosis of retinal cells,thereby maintaining the integrity of the blood-retina barrier.
作者
崔家霖
邢俊艳
王莺洁
高彦彬
CUI Jia-Lin;XING Jun-Yan;WANG Ying-Jie;GAO Yan-Bin(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China;Beijing Key Laboratory of Collateral Diseases of Traditional Chinese Medicine,Beijing 100069,China)
出处
《广州中医药大学学报》
CAS
2022年第3期605-611,共7页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家重点基础研究发展计划(973计划)项目(编号:2018YFC1704102)。
