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目的探究活血方提取物(EBRE)对2型糖尿病GK大鼠视网膜病变保护机制。方法将28周龄GK大鼠随机分为模型组、EBRE低、中、高剂量组和阳性对照组,每组各5只,另取5只Wistar大鼠为阴性对照组。EBRE低、中、高剂量组分别灌胃予以3.90,7.80,15.60 g·kg~(-1)EBRE,阳性对照组灌胃予以150.00 mg·kg~(-1)羟苯磺酸钙,模型组和阴性对照组均灌胃予以等体积0.9%NaCl。6组大鼠每天灌胃一次,连续给药12周。给药4,8,12周后,检测6组大鼠的空腹血糖(FBG);给药12周后,用HE染色法检测视网膜厚度和视网膜神经节细胞(RGCs)计数,用免疫染色法检测大鼠视网膜β-catenin、环氧合酶-2(COX-2)和血管内皮生长因子(VEGF)表达。结果给药12周后,模型组、阴性对照组、阳性对照组、EBRE低、中、高剂量组的FBG分别为(7.66±0.68),(4.35±0.75),(5.77±0.81),(5.50±0.61),(5.98±0.70),(6.25±0.29)mmol·L~(-1);视网膜厚度分别为(38.07±7.57)%,(92.50±26.15)%,(60.11±13.56)%,(32.86±15.06)%,(78.32±11.88)%,(47.03±9.76)%;RGCs计数分别为(47.08±5.84)%,(100.00±21.91)%,(66.65±13.95)%,(59.19±5.96)%,(82.90±9.69)%,(61.69±9.10)%;β-catenin表达量分别为(295.35±56.63)%,(100.00±34.44)%,(268.65±52.65)%,(190.10±43.90)%,(227.66±53.92)%,(298.29±57.79)%;COX-2表达量分别为(186.40±78.86)%,(100.00±39.94)%,(78.29±26.99)%,(57.21±13.92)%,(55.11±14.99)%,(203.62±50.22)%;VEGF表达量分别为(381.59±146.52)%,(100.00±24.26)%,(211.48±68.18)%,(193.36±71.10)%,(195.63±44.60)%,(308.89±125.60)%,模型组与阴性对照组比较差异均有统计学意义(P<0.05)。所有数据除FBG外都以阴性对照为100.00%。结论 EBRE通过降低GK大鼠FBG,抑制视网膜β-catenin表达,减少RGCs丢失,改善视网膜病变。
Objective To investigate the protective mechanism of huoxuefang extract (EBRE) on retinopathy of type 2 diabetic GK rats. Methods 28-week-old GK rats were randomly divided into model group, EBRE low, medium and high dose groups and positive control group, each with 5 rats and another 5 Wistar rats as negative control group. EBRE low, medium and high dose groups were intragastrically administered 3.90,7.80,15.60 g · kg -1 EBRE, the positive control group were given 150.00 mg · kg -1 calcium dobesilate, the model group And negative control group were given an equal volume of 0.9% NaCl. Six groups of rats were given gavage once daily for 12 weeks. Fasting plasma glucose (FBG) was measured in rats of 4, 8 and 12 weeks after administration. After 12 weeks of administration, retinal ganglion cells (RGCs) and retinal ganglion cells (RGCs) were counted by HE staining and detected by immunostaining Expression of β-catenin, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) in rat retina. Results The FBG of model group, negative control group, positive control group and EBRE low, medium and high dose groups were (7.66 ± 0.68), (4.35 ± 0.75), (5.77 ± 0.81), (5.50 The retinal thickness were (38.07 ± 7.57)%, (92.50 ± 26.15)% and (60.11 ± 13.56)%, respectively. The thickness of the retina was (± 0.61), (5.98 ± 0.70) and (6.25 ± 0.29) mmol·L -1 The average counts of RGCs were (47.08 ± 5.84)%, (100.00 ± 21.91)%, (66.65 ± 13.95)% and (59.19 ± 5.96)%, respectively The percentages of β-catenin expression were (295.35 ± 56.63)%, (100.00 ± 34.44)%, (268.65 ± 52.65)%, (190.10 ± 43.90)%, respectively , (227.66 ± 53.92)% and (298.29 ± 57.79)%, respectively. The expression of COX-2 in the two groups were (186.40 ± 78.86)%, (100.00 ± 39.94)%, (78.29 ± 26.99)% and (57.21 ± 13.92) (55.11 ± 14.99)% and (203.62 ± 50.22)%, respectively. The expression of VEGF was (381.59 ± 146.52)%, (100.00 ± 24.26)%, (211.48 ± 68.18)%, (193.36 ± 71.10)% and 44.60)%, (308.89 ± 125.60)% respectively. The differences between the model group and the negative control group were statistically significant (P <0.05). All data except for FBG negative control was 100.00%. Conclusion EBRE can reduce retinal β-catenin expression, reduce RGCs loss and improve retinopathy by reducing FBG in GK rats.