补肾活血中药对早期糖尿病大鼠视网膜内屏障损伤的影响
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摘要
目的探索补肾活血中药复方对高脂高糖饮食联合STZ诱导大鼠早期发生糖尿病视网膜内屏障(i BRB)损伤模型的影响。方法高糖高脂饮食和STZ诱导对标准大鼠进行造模,随机将早期糖尿病i BRB损伤的大鼠模型分为5个组,分别是空白模型组、阳性对照组、高中低3种剂量中药组,另设正常对照组。6组均在1周、2周、4周、8周后观察并设相应亚组。对大鼠的活体进行灌注并固定,行视网膜血管消化铺片,观察并记录大鼠视网膜血管的形态学变化。视网膜屏障渗漏情况通过异硫氰酸罗丹明(RHIC)检测。结果各组大鼠第8周时间亚组的糖尿病SD大鼠视网膜形态学未见明显改变。在各组糖尿病SD大鼠的第2周亚组、4周亚组、8周亚组RHIC渗漏主要出现在大鼠视网膜的外丛状层、内核层、内丛状层、神经节细胞层以及神经纤维层,且外丛状层RHIC渗漏情况最为明显,其中以模型组RHIC渗漏最多,中、高剂量中药组RHIC渗漏情况相近且仅次于模型组,低剂量中药组RHIC渗漏量低于中、高剂量中药组,阳性对照组RHIC渗漏最少。结论糖尿病SD大鼠在患病后8周内的视网膜形态不会发生显著改变。i BRB损害从第2周开始出现并逐渐加重。补肾活血中药复方对糖尿病视网膜病变大鼠i BRB损伤后渗漏有一定缓解作用。
OBJECTIVE To investigate the high glucose and high fat diet and Streptozotocin(STZ) induced early-stage diabetic internal blood-retinal barrier(iBRB) damage rat model and the influence of invigorating kidney and activating blood formula on i BRB leakage. METHODS High glucose and high fat diet and STZ rats were used to induce rat model. After the successful modeling, all rats were randomly divided into model group, positive control group, high, medium and low dose Chinese herbal compound groups, and normal SD rats were set as the normal control group.Each group was observed in the 1 st, 2 nd, 4 th, and 8 th week. Then the rats were prepared for vivi-perfusion and fixation, their retinal vessels were used for stretched preparation and the morphological changes were observed. The RHIC was used to detect the leakage of retinal barrier.RESULTS At the 8 th week, the retinal morphology of DM rats showed no obvious change. The diabetes rats' retinal blood vessels were RHIC leakage at the 2 nd,4 th, and 8 th week, leakage was located within the outer plexiform layer, the inner plexiform layer, inner nuclear layer, ganglion cell layer and nerve fiber layer while outer plexiform l ayer RHIC leakage was most significant. Specifically, RHIC leakage was the most in the negative control group, RHIC leakage in the medium and high dose Chinese medicine group was similar to that in the negative control group, while RHIC leakage in the low dose Chinese medicine group was lower than that in the medium and high dose Chinese medicine group, and RHIC leakage in the positive control group was the least. CONCLUSIONS Diabetic SD rats showed no significant changes in retinal morphology within 8 weeks after disease onset. The iBRB damage began to appear from the second week and gradually worsened. The traditional Chinese medicine compound for tonifying kidney and promoting blood circulation had a certain alleviating effect on i BRB leakage after diabetic retinopathy injury in rats.
OBJECTIVE To investigate the high glucose and high fat diet and Streptozotocin(STZ) induced early-stage diabetic internal blood-retinal barrier(iBRB) damage rat model and the influence of invigorating kidney and activating blood formula on i BRB leakage. METHODS High glucose and high fat diet and STZ rats were used to induce rat model. After the successful modeling, all rats were randomly divided into model group, positive control group, high, medium and low dose Chinese herbal compound groups, and normal SD rats were set as the normal control group.Each group was observed in the 1 st, 2 nd, 4 th, and 8 th week. Then the rats were prepared for vivi-perfusion and fixation, their retinal vessels were used for stretched preparation and the morphological changes were observed. The RHIC was used to detect the leakage of retinal barrier.RESULTS At the 8 th week, the retinal morphology of DM rats showed no obvious change. The diabetes rats' retinal blood vessels were RHIC leakage at the 2 nd,4 th, and 8 th week, leakage was located within the outer plexiform layer, the inner plexiform layer, inner nuclear layer, ganglion cell layer and nerve fiber layer while outer plexiform l ayer RHIC leakage was most significant. Specifically, RHIC leakage was the most in the negative control group, RHIC leakage in the medium and high dose Chinese medicine group was similar to that in the negative control group, while RHIC leakage in the low dose Chinese medicine group was lower than that in the medium and high dose Chinese medicine group, and RHIC leakage in the positive control group was the least. CONCLUSIONS Diabetic SD rats showed no significant changes in retinal morphology within 8 weeks after disease onset. The iBRB damage began to appear from the second week and gradually worsened. The traditional Chinese medicine compound for tonifying kidney and promoting blood circulation had a certain alleviating effect on i BRB leakage after diabetic retinopathy injury in rats.
引文
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[11]施沃栋,王志良,罗敏,等.链脲佐菌素诱发糖尿病大鼠模型视网膜形态学改变的观察[J].眼科新进展,2008,28(8):583-585,589.
[12] ZHANG J,WU Y,JIN Y,et al. Intravitreal injection of erythropoietin protects both retinal vascular and neuronal cells in early diabetes[J].Invest Ophthalmol Vis Sci,2008,49(2):732-742.
[13] ZENG XX,NG YK,LING EA. Neuronal and microglial response in the retina of streptozotocin-induced diabetic rats[J].Visual Neuroscience,2000,17(3):463-471.
[14] DUH EJ,YANG HS,HALLER JA,et al.Vitreous levels of pigment epithelium-derived factor and vascular endothelial growth factor:implications for ocular angiogenesis[J].Am J Ophthalmol,2004,137(4):668-674.
[15] ZHANG SX,MA JX,SIMA J,et al.Genetic difference in susceptibility to the blood-retina barrier breakdown in diabetes and oxygeninduced retinopathy[J].Am J Pathol,2005,166(1):313-321.
[16] TOLENTINO MJ,MCLEOD DS,TAOMOTO M,et al.Pathologic features of vascular endothelial growth factor-induced retinopathy in the nonhuman primate[J].Am J Ophthalmol,2002,133(3):373-385.
[17]李艳,李筱荣,袁佳琴,等.糖尿病大鼠视网膜中VEGF、PEDF的表达与血-视网膜屏障损伤[J].眼科新进展,2013,33(1):29-32.
[2] LOKMIC Z,MUSYOKA J,HEWITSON T D,et al. Hypoxia and hypoxia signaling in tissue repair and fibrosis.[J]. Int Rev Cell Mol Biol,2012,296:139-85.
[3]朱琦,夏欣,许迅,等.蛋白激酶C与糖尿病视网膜病变血.视网膜屏障破坏关系的实验研究[J].中华眼科杂志.2004,40(8):565-566.
[4]陈永东,许迅.糖尿病血-视网膜屏障破坏的机制研究进展[J].2009,33(5):357-360.
[5] ANTCLIFF RJ,MARSHALL J.The pathogenesis of edema in diabetic maculopathy[J].Semin Ophthaimol,1999,14(4):223-232.
[6] LI W, SHEN S,KHATAMI M,et al. Stimulation of retinal capillary pericyte protein and collagen synthesis in culture by high-glucose concentration[J].Diabetes,1984,33(8):785-789.
[7]庞东渤,刘学政,符丽娟.细胞凋亡与STZ糖尿病大鼠视网膜微血管病变关系[J].眼科新进展,2006,26(2):111-113.
[8]王康,刘乃慧,李新民.两种糖尿病大鼠模型血-视网膜屏障对EB通透性的比较[J].眼科新进展,2007,27(1):22-24.
[9] ANDERSON HR, STITT AW, GARDINER TA, et al. Diabetic retinopathy:morphometric analysis of basement membrane thickening of capillaries in different retinal layers within arterial and venous environments[J].Br J Ophthalmol,1995,79(12):1120-1123.
[10] GASTINGER MJ,SINGH RSJ,BARBER AJ. Loss of cholinergic and dopaminergic amacrine cells in streptozotocin-diabetic rat and ins2akita-diabetic mouse retinas[J].Invest Ophthalmol Vis Sci,2006,47(7):3143-3150.
[11]施沃栋,王志良,罗敏,等.链脲佐菌素诱发糖尿病大鼠模型视网膜形态学改变的观察[J].眼科新进展,2008,28(8):583-585,589.
[12] ZHANG J,WU Y,JIN Y,et al. Intravitreal injection of erythropoietin protects both retinal vascular and neuronal cells in early diabetes[J].Invest Ophthalmol Vis Sci,2008,49(2):732-742.
[13] ZENG XX,NG YK,LING EA. Neuronal and microglial response in the retina of streptozotocin-induced diabetic rats[J].Visual Neuroscience,2000,17(3):463-471.
[14] DUH EJ,YANG HS,HALLER JA,et al.Vitreous levels of pigment epithelium-derived factor and vascular endothelial growth factor:implications for ocular angiogenesis[J].Am J Ophthalmol,2004,137(4):668-674.
[15] ZHANG SX,MA JX,SIMA J,et al.Genetic difference in susceptibility to the blood-retina barrier breakdown in diabetes and oxygeninduced retinopathy[J].Am J Pathol,2005,166(1):313-321.
[16] TOLENTINO MJ,MCLEOD DS,TAOMOTO M,et al.Pathologic features of vascular endothelial growth factor-induced retinopathy in the nonhuman primate[J].Am J Ophthalmol,2002,133(3):373-385.
[17]李艳,李筱荣,袁佳琴,等.糖尿病大鼠视网膜中VEGF、PEDF的表达与血-视网膜屏障损伤[J].眼科新进展,2013,33(1):29-32.