苦参碱对糖尿病视网膜病变大鼠视网膜微血管内皮细胞增生及血管内皮生长因子表达的影响
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摘要
目的:探讨苦参碱对糖尿病视网膜病变大鼠视网膜微血管内皮细胞增生及血管内皮生长因子(VEGF)表达的影响。方法:将90只SD大鼠随机分为正常组、模型组、对照组、低剂量组、中剂量组及高剂量组,每组15只。正常组大鼠常规饲养,其余组大鼠制作糖尿病大鼠模型。对照组大鼠给予羟苯磺酸钙30 mg/(kg·d),灌胃;低、中及高剂量组大鼠分别给予苦参碱30、60及90 mg/(kg·d),灌胃;正常组、模型组大鼠给予等量的蒸馏水。连续给药20周后,检测大鼠血清超氧化物歧化酶1(SOD1)、丙二醛(MDA)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和C反应蛋白(CRP)含量,观察视网膜组织的病理切片,检测VEGF、促血管生成素1(ANG1)蛋白的表达。结果:(1)大鼠视网膜组织病理切片显示,低剂量组、中剂量组及高剂量组大鼠的视网膜细胞水肿症状好于对照组,且随着苦参碱剂量的升高,细胞结构完整性提高,扩张的微血管更少。(2)低剂量组、中剂量组及高剂量组大鼠视网膜微血管内皮细胞凋亡数明显高于对照组,差异均有统计学意义(P<0.05),且随着苦参碱剂量的升高,凋亡数增多。(3)低剂量组、中剂量组及高剂量组大鼠SOD1水平明显高于模型组和对照组,MDA、CRP水平明显低于模型组和对照组,差异均有统计学意义(P<0.05),且随着苦参碱剂量的升高,SOD1水平升高,MDA、CRP水平降低。(4)低剂量组、中剂量组及高剂量组大鼠TNF-α、IL-1β及IL-6水平明显低于模型组和对照组,差异均有统计学意义(P<0.05),且随着苦参碱剂量的升高,TNF-α、IL-1β及IL-6水平降低。(5)低剂量组、中剂量组及高剂量组大鼠视网膜内VEGF、ANG1蛋白含量明显低于模型组和对照组,差异均有统计学意义(P<0.05),且随着苦参碱剂量的升高,视网膜内VEGF、ANG1蛋白含量降低。结论:苦参碱可改善糖尿病大鼠的视网膜损伤,抑制视网膜微血管内皮细胞的增生;其是通过调节VEGF、ANG1的表达,提高SOD1的表达,减少MDA、CRP、TNF-α、IL-1β及IL-6含量来发挥改善病情的作用。
OBJECTIVE: To probe into the effect of matrine on proliferation of retinal microvascular endothelial cell and expression of vascular endothelial growth factor(VEGF) in rats with diabetic retinopathy. METHODS: 90 SD rats were randomly divided into normal group, model group, control group, low-dose group, medium-dose group and high-dose group, with 15 rats in each group. The normal group was routinely feeding, while the rest were made into diabetic rat model. The control group was given 30 mg/(kg·d) of calcium dobesilate and intragastric administration; the low-dose group, medium-dose group and high-dose group were respectively given matrine 30, 60 and 90 mg/(kg·d), with intragastric administration also; the normal group and model group were given equal amount of distilled water. After continuous administration for 20 weeks, the rats were tested for serum superoxide dismutase 1(SOD1), malondialdehyde(MDA), tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6) and C-reactive protein(CRP), the pathological sections of retinal tissues were observed to detect the expression of VEGF and angiopoietin 1(ANG1) protein. RESULTS:(1) Pathological sections of rat retina indicated that the symptoms of retinal cell edema in low-dose group, medium-dose group and high-dose group were better than that of control group, as the increase of matrine dose, the cell structural integrity was improved and expansion of microvascular was less.(2) The number of retinal microvascular endothelial cell apoptosis in low-dose group, medium-dose group and high-dose group were significantly higher than those of the control group, with statistically significant differences(P<0.05), as the increse of matrine dose, the number of apoptosis was also increased.(3) The SOD1 levels in low-dose group, medium-dose group and high-dose group were significantly higher than those of the model group and control group, the MDA and CRP levels were significantly lower than those of the model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the SOD1 levels was also increased, while the MDA and CRP levels were decreased.(4) The TNF-α, IL-1β and IL-6 levels of low-dose group, medium-dose group and high-dose group were significantly lower than those of the model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the TNF-α, IL-1β and IL-6 levels were also decreased.(5) The VEGF and ANG1 protein contents of low-dose group, medium-dose group and high-dose group were significantly lower than those of model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the VEGF and ANG1 protein contents were decreased. CONCLUSIONS: Matrine can improve retinal damage and inhibit the proliferation of retinal microvascular endothelial cells in diabetic rats; it's supposed that by regulating the expression of VEGF and ANG1, the expression of SOD1 is improved, the contents of MDA, CRP, TNF-α and IL-1β are reduced to play the role of improving illness.
OBJECTIVE: To probe into the effect of matrine on proliferation of retinal microvascular endothelial cell and expression of vascular endothelial growth factor(VEGF) in rats with diabetic retinopathy. METHODS: 90 SD rats were randomly divided into normal group, model group, control group, low-dose group, medium-dose group and high-dose group, with 15 rats in each group. The normal group was routinely feeding, while the rest were made into diabetic rat model. The control group was given 30 mg/(kg·d) of calcium dobesilate and intragastric administration; the low-dose group, medium-dose group and high-dose group were respectively given matrine 30, 60 and 90 mg/(kg·d), with intragastric administration also; the normal group and model group were given equal amount of distilled water. After continuous administration for 20 weeks, the rats were tested for serum superoxide dismutase 1(SOD1), malondialdehyde(MDA), tumor necrosis factor α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6) and C-reactive protein(CRP), the pathological sections of retinal tissues were observed to detect the expression of VEGF and angiopoietin 1(ANG1) protein. RESULTS:(1) Pathological sections of rat retina indicated that the symptoms of retinal cell edema in low-dose group, medium-dose group and high-dose group were better than that of control group, as the increase of matrine dose, the cell structural integrity was improved and expansion of microvascular was less.(2) The number of retinal microvascular endothelial cell apoptosis in low-dose group, medium-dose group and high-dose group were significantly higher than those of the control group, with statistically significant differences(P<0.05), as the increse of matrine dose, the number of apoptosis was also increased.(3) The SOD1 levels in low-dose group, medium-dose group and high-dose group were significantly higher than those of the model group and control group, the MDA and CRP levels were significantly lower than those of the model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the SOD1 levels was also increased, while the MDA and CRP levels were decreased.(4) The TNF-α, IL-1β and IL-6 levels of low-dose group, medium-dose group and high-dose group were significantly lower than those of the model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the TNF-α, IL-1β and IL-6 levels were also decreased.(5) The VEGF and ANG1 protein contents of low-dose group, medium-dose group and high-dose group were significantly lower than those of model group and control group, with statistically significant differences(P<0.05), as the increase of matrine dose, the VEGF and ANG1 protein contents were decreased. CONCLUSIONS: Matrine can improve retinal damage and inhibit the proliferation of retinal microvascular endothelial cells in diabetic rats; it's supposed that by regulating the expression of VEGF and ANG1, the expression of SOD1 is improved, the contents of MDA, CRP, TNF-α and IL-1β are reduced to play the role of improving illness.
引文
[1] 王春雁.糖尿病视网膜病变患者血清VEGF、NO的变化及临床意义[J].中国实验诊断学,2015,19(1):94-95.
[2] 于静,张明昌.苦参碱对大鼠角膜新生血管的抑制作用[J].眼科新进展,2012,32(6):532-535.
[3] 邓志华,黄赞松,黎梨等.苦参素对人肝癌Bel-7404细胞增殖及E2F1、c-myc基因表达的影响[J].广西医科大学学报,2018,35(9):1191-1194.
[4] 夏世刚,彭辉灿.密蒙花提取物对高糖下人视网膜血管内皮细胞增生及VEGF表达的影响[J].中华实验眼科杂志,2011,29(9):798-799.
[5] 姚昱欧,梁建宏,黎晓新.微切口玻璃体视网膜手术治疗增生型糖尿病视网膜病变合并牵拉性视网膜脱离的疗效观察[J].中华眼底病杂志,2016,32(5):495-499.
[6] 李晓萍.血管内皮生长因子与糖尿病微血管并发症的关系[J].中国综合临床,2011,27(2):222-224.
[7] 韩林峰,柯根杰,王林,等.全视网膜激光光凝对增生型糖尿病视网膜病变视网膜前膜中环氧化酶-2、血管内皮生长因子表达的影响[J].中华眼底病杂志,2016,32(2):140-143.
[8] 路强,杨晓静,崔巍,等.内脏脂肪素和血管内皮生长因子在早期糖尿病大鼠视网膜中的表达及意义[J].中华实验眼科杂志,2013,31(1):45-48.
[9] 邢立臣,张雷鸣,丰亚丽,等.血清中内皮细胞生长因子、白细胞介素-1及肿瘤坏死因子-α水平与老年糖尿病视网膜病变的关系[J].中国老年学杂志,2016,36(12):2991-2992.
[10] 段娜,戴丹,周灵.血栓通联合眼底激光对糖尿病视网膜病变黄斑水肿患者血清NOS、VEGF和IL-6的影响[J].河北医药,2016,38(18):2751-2753,2757.
[11] 温积权,汪怿,杨杰,等.糖尿病性视网膜病变患者VEGF与微血管损伤的相关性[J].国际眼科杂志,2015,15(12):2063-2066.
[12] 徐家窈,冼文光,刘俐娜.血清VEGF及ICAM-1水平与糖尿病视网膜病变患者微血管损伤的关系[J].中国老年学杂志,2017,37(8):2000-2002.
[13] 王彦方,蓝淑琴,李霞,等.糖脉康颗粒联合羟苯磺酸钙治疗糖尿病视网膜病变的疗效及对血清炎症因子的影响[J].中国慢性病预防与控制,2018,26(9):673-676.
[14] 徐寒松,孔德明,向慧.通脉糖眼明胶囊对糖尿病视网膜病变患者血管内皮生长因子水平的影响[J].第三军医大学学报,2011,33(13):1327,1332.
[15] 李宜川,母传贤,沈永杰等.糖尿病视网膜病变与血清标志物的相关性研究[J].现代预防医学,2012,39(13):3404-3405,3408.
[2] 于静,张明昌.苦参碱对大鼠角膜新生血管的抑制作用[J].眼科新进展,2012,32(6):532-535.
[3] 邓志华,黄赞松,黎梨等.苦参素对人肝癌Bel-7404细胞增殖及E2F1、c-myc基因表达的影响[J].广西医科大学学报,2018,35(9):1191-1194.
[4] 夏世刚,彭辉灿.密蒙花提取物对高糖下人视网膜血管内皮细胞增生及VEGF表达的影响[J].中华实验眼科杂志,2011,29(9):798-799.
[5] 姚昱欧,梁建宏,黎晓新.微切口玻璃体视网膜手术治疗增生型糖尿病视网膜病变合并牵拉性视网膜脱离的疗效观察[J].中华眼底病杂志,2016,32(5):495-499.
[6] 李晓萍.血管内皮生长因子与糖尿病微血管并发症的关系[J].中国综合临床,2011,27(2):222-224.
[7] 韩林峰,柯根杰,王林,等.全视网膜激光光凝对增生型糖尿病视网膜病变视网膜前膜中环氧化酶-2、血管内皮生长因子表达的影响[J].中华眼底病杂志,2016,32(2):140-143.
[8] 路强,杨晓静,崔巍,等.内脏脂肪素和血管内皮生长因子在早期糖尿病大鼠视网膜中的表达及意义[J].中华实验眼科杂志,2013,31(1):45-48.
[9] 邢立臣,张雷鸣,丰亚丽,等.血清中内皮细胞生长因子、白细胞介素-1及肿瘤坏死因子-α水平与老年糖尿病视网膜病变的关系[J].中国老年学杂志,2016,36(12):2991-2992.
[10] 段娜,戴丹,周灵.血栓通联合眼底激光对糖尿病视网膜病变黄斑水肿患者血清NOS、VEGF和IL-6的影响[J].河北医药,2016,38(18):2751-2753,2757.
[11] 温积权,汪怿,杨杰,等.糖尿病性视网膜病变患者VEGF与微血管损伤的相关性[J].国际眼科杂志,2015,15(12):2063-2066.
[12] 徐家窈,冼文光,刘俐娜.血清VEGF及ICAM-1水平与糖尿病视网膜病变患者微血管损伤的关系[J].中国老年学杂志,2017,37(8):2000-2002.
[13] 王彦方,蓝淑琴,李霞,等.糖脉康颗粒联合羟苯磺酸钙治疗糖尿病视网膜病变的疗效及对血清炎症因子的影响[J].中国慢性病预防与控制,2018,26(9):673-676.
[14] 徐寒松,孔德明,向慧.通脉糖眼明胶囊对糖尿病视网膜病变患者血管内皮生长因子水平的影响[J].第三军医大学学报,2011,33(13):1327,1332.
[15] 李宜川,母传贤,沈永杰等.糖尿病视网膜病变与血清标志物的相关性研究[J].现代预防医学,2012,39(13):3404-3405,3408.