渴络欣胶囊对db/db小鼠早期糖尿病视网膜病的防治作用研究
|
摘要
探讨渴络欣胶囊对db/db小鼠早期糖尿病视网膜病(diabetic retinopathy,DR)的改善作用。采用目前国际公认的8周龄雄性(45～55 g)db/db自发性糖尿病小鼠动物模型,每周称取体质量和空腹血糖1次,根据体质量灌胃给药。渴络欣低、中、高剂量组灌胃剂量分别为780,1 560,3 120 mg·kg~(-1)·d~(-1);羟苯磺酸钙组灌胃剂量为195 mg·kg~(-1)·d~(-1);对照组和模型组均灌胃相等体积生理盐水;各组连续给药20周。灌胃结束后,进行视网膜光学相干断层扫描,眼底荧光血管造影和视网膜电图。小鼠摘取眼球,分离视网膜,制作石蜡切片,HE染色;免疫组化检测视网膜胶质纤维酸性蛋白表达。渴络欣胶囊对db/db小鼠体质量和空腹血糖并无显著调节作用。相干断层实验结果表明,渴络欣胶囊能够显著改善小鼠视网膜神经节层和内丛状层的厚度;眼底荧光造影结果表明,渴络欣胶囊能够一定程度改善早期糖尿病视网膜病微血管数量;ERG实验表明渴络欣胶囊能够改善DR早期视网膜功能。视网膜组织病理切片染色证实渴络欣胶囊能够一定程度改善各层细胞排列不规则、水肿,免疫组化实验结果显示各组视网膜组织并未出现明显的胶质化。该研究结果表明,渴络欣胶囊能够显著降低db/db糖尿病小鼠视网膜神经节层和内丛状层的厚度;改善DR早期视网膜功能;改善视网膜各层水肿、排列紊乱等。提示渴络欣胶囊能够防治早期糖尿病视网膜病,这为临床上渴络欣治疗糖尿病视网膜病提供了实验依据。
The aim of this paper was to investigate the preventive effects of Keluoxin Capsules(KLX) on diabetic retinopathy in db/db mice. One hundred male db/db diabetic mice(45-55 g, 8 weeks) were randomly divided into 5 groups(model, KLX low dose, KLX middle dose, KLX high dose, Dobesilate) and 20 male C57 BL/KsJdb~(+/+) were taken as control group. Body weight and fasting blood-glucose were detected every week. Mice were administrated with saline(control and model group), KLX(780, 1 560, 3 120 mg·kg~(-1)·d~(-1), ig), Dobesilate(195 mg·kg~(-1)·d~(-1), ig) for 20 weeks, respectively. At the end of the administration, optical coherence tomography, fundus fluorescein angiography and electroretinogram of the retina were measured. The eyeball was extirpated and retina was isolated to make paraffin section, followed by HE staining and glial fibrillary acidic protein(GFAP) immunohistochemistry. The results indicated that KLX has no obvious effect on body weight and fasting blood level in db/db mice. However, KLX could significantly regulate the thickness of retinal ganglion layer and inner plexiform layer. KLX was able to remarkably reduce the quantity of diabetic microvessel. Meanwhile, KLX could notably improve retinal function. Moreover, KLX could observably modulate the cell arrangement and edema in each layer. There was no markable difference in retina according to the immunochemistry assay. In the present study, KLX exert marked preventive effects on diabetic retinopathy in db/db mice, which provided an experimental evidence for clinical use.
The aim of this paper was to investigate the preventive effects of Keluoxin Capsules(KLX) on diabetic retinopathy in db/db mice. One hundred male db/db diabetic mice(45-55 g, 8 weeks) were randomly divided into 5 groups(model, KLX low dose, KLX middle dose, KLX high dose, Dobesilate) and 20 male C57 BL/KsJdb~(+/+) were taken as control group. Body weight and fasting blood-glucose were detected every week. Mice were administrated with saline(control and model group), KLX(780, 1 560, 3 120 mg·kg~(-1)·d~(-1), ig), Dobesilate(195 mg·kg~(-1)·d~(-1), ig) for 20 weeks, respectively. At the end of the administration, optical coherence tomography, fundus fluorescein angiography and electroretinogram of the retina were measured. The eyeball was extirpated and retina was isolated to make paraffin section, followed by HE staining and glial fibrillary acidic protein(GFAP) immunohistochemistry. The results indicated that KLX has no obvious effect on body weight and fasting blood level in db/db mice. However, KLX could significantly regulate the thickness of retinal ganglion layer and inner plexiform layer. KLX was able to remarkably reduce the quantity of diabetic microvessel. Meanwhile, KLX could notably improve retinal function. Moreover, KLX could observably modulate the cell arrangement and edema in each layer. There was no markable difference in retina according to the immunochemistry assay. In the present study, KLX exert marked preventive effects on diabetic retinopathy in db/db mice, which provided an experimental evidence for clinical use.
引文
[1] International Diabetes Federation.IDF Diabetes Atlas 8th Edition[EB/OL].[2018-12-10].https://www.idf.org/our-activities/advocacy-awareness/resources-and-tools/134:idf-diabetes-atlas-8th-edition.html.
[2] Antonetti D A,Klein R,Gardner T W.Diabetic retinopathy[J].N Engl J Med,2012,366(13):1227.
[3] Sohn E H,van Dijk H W,Jiao C,et al.Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus[J].Proc Natl Acad Sci U S A,2016,113(19):E2655.
[4] American Diabetes Association.10.Microvascular complications and foot care:standards of medical care in diabetes-2018[J].Diabetes Care,2018,41(Suppl 1):S105.
[5] 赵豫梅,吴于滨,宋滇平.渴络欣胶囊联合ARB 类药物治疗糖尿病肾病有效性及安全性的系统评价[J].中国中西医结合肾病杂志,2017,18(7):610.
[6] 胡金芳,王玮,吴建明,等.渴络欣胶囊对缺氧诱导视网膜血管新生的抑制作用[J].中国实验方剂学杂志,2014,20(10):156.
[7] 申秀萍,马娜,吴建明,等.渴络欣胶囊对STZ诱导糖尿病大鼠视网膜病变的防治作用[J].中国实验方剂学杂志,2014,20(12):188.
[8] 赵洁,莫超,孟立锋,等.补阳还五汤治疗早期糖尿病肾病的疗效及安全性的Meta分析[J].中国中药杂志,2019,44(8):1660.
[9] 熊兴江.基于临床重症病例的麻黄附子细辛汤方证特征及其治疗脓毒症、多脏器功能衰竭、气管插管术后高热不退、急性心梗合并糖尿病末梢神经病变剧烈疼痛研究[J].中国中药杂志,2019,doi:10.19540/j.cnki.cjcmm.20190416.503.
[10] 白宇,贺云,杨丽霞.活血化瘀类中药治疗糖尿病肾病机制的研究进展[J].中国实验方剂学杂志,2018,24(23):200.
[11] 萧韵健.渴络欣治疗非增殖性糖尿病视网膜病变的临床观察[D].武汉:湖北中医药大学,2015.
[12] 冀向宁,刘玉青,韩风梅,等.OCT与FFA在糖尿病视网膜病变临床前期的应用比较[J].眼科新进展,2014,34(7):662.
[13] 张季瑾.糖尿病视网膜病变患者黄斑厚度OCT测量定量分析[J].河北医学,2013,19(5):738.
[14] 马宏杰,李月灵,郑燕林.双参明目片对非增殖期糖尿病视网膜病变患者视网膜功能的干预作用[J].中国医院药学,2016(36):260.
[15] Ning Cheung,Paul Mitchell,Tien Yin Wong.Diabetic retinopathy[J].Lancet,2010,376(9735):124.
[16] Cogan D G,Toussaint D,Kuwabara T.Retinal vascular patterns.Ⅳ.Diabetic retinopathy[J].Arch Ophthalmol,1961,66:366.
[17] 于波.光学相干断层扫描与眼底荧光血管造影在糖尿病视网膜病变临床前期的应用价值[J].中国药物经济学,2015,10(12):192.
[18] Sohn E H,van Dijk H W,Jiao C,et al.Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus[J].Proc Natl Acad Sci U S A,2016,113(19):E2655.
[19] 李芳,张晨,陈雪艺,等.闪光视觉诱发电位联合闪光视网膜电流图检查评价白内障患者术后视功能[J].中国老年学杂志,2013,33(23):5858.
[20] 徐寒松,孔德明,李雪梅,等.通脉糖眼明胶囊对单纯型糖尿病视网膜病变患者闪光视网膜电图及其振荡电位的影响[J].中医杂志,2007,48(6):512.
[2] Antonetti D A,Klein R,Gardner T W.Diabetic retinopathy[J].N Engl J Med,2012,366(13):1227.
[3] Sohn E H,van Dijk H W,Jiao C,et al.Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus[J].Proc Natl Acad Sci U S A,2016,113(19):E2655.
[4] American Diabetes Association.10.Microvascular complications and foot care:standards of medical care in diabetes-2018[J].Diabetes Care,2018,41(Suppl 1):S105.
[5] 赵豫梅,吴于滨,宋滇平.渴络欣胶囊联合ARB 类药物治疗糖尿病肾病有效性及安全性的系统评价[J].中国中西医结合肾病杂志,2017,18(7):610.
[6] 胡金芳,王玮,吴建明,等.渴络欣胶囊对缺氧诱导视网膜血管新生的抑制作用[J].中国实验方剂学杂志,2014,20(10):156.
[7] 申秀萍,马娜,吴建明,等.渴络欣胶囊对STZ诱导糖尿病大鼠视网膜病变的防治作用[J].中国实验方剂学杂志,2014,20(12):188.
[8] 赵洁,莫超,孟立锋,等.补阳还五汤治疗早期糖尿病肾病的疗效及安全性的Meta分析[J].中国中药杂志,2019,44(8):1660.
[9] 熊兴江.基于临床重症病例的麻黄附子细辛汤方证特征及其治疗脓毒症、多脏器功能衰竭、气管插管术后高热不退、急性心梗合并糖尿病末梢神经病变剧烈疼痛研究[J].中国中药杂志,2019,doi:10.19540/j.cnki.cjcmm.20190416.503.
[10] 白宇,贺云,杨丽霞.活血化瘀类中药治疗糖尿病肾病机制的研究进展[J].中国实验方剂学杂志,2018,24(23):200.
[11] 萧韵健.渴络欣治疗非增殖性糖尿病视网膜病变的临床观察[D].武汉:湖北中医药大学,2015.
[12] 冀向宁,刘玉青,韩风梅,等.OCT与FFA在糖尿病视网膜病变临床前期的应用比较[J].眼科新进展,2014,34(7):662.
[13] 张季瑾.糖尿病视网膜病变患者黄斑厚度OCT测量定量分析[J].河北医学,2013,19(5):738.
[14] 马宏杰,李月灵,郑燕林.双参明目片对非增殖期糖尿病视网膜病变患者视网膜功能的干预作用[J].中国医院药学,2016(36):260.
[15] Ning Cheung,Paul Mitchell,Tien Yin Wong.Diabetic retinopathy[J].Lancet,2010,376(9735):124.
[16] Cogan D G,Toussaint D,Kuwabara T.Retinal vascular patterns.Ⅳ.Diabetic retinopathy[J].Arch Ophthalmol,1961,66:366.
[17] 于波.光学相干断层扫描与眼底荧光血管造影在糖尿病视网膜病变临床前期的应用价值[J].中国药物经济学,2015,10(12):192.
[18] Sohn E H,van Dijk H W,Jiao C,et al.Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus[J].Proc Natl Acad Sci U S A,2016,113(19):E2655.
[19] 李芳,张晨,陈雪艺,等.闪光视觉诱发电位联合闪光视网膜电流图检查评价白内障患者术后视功能[J].中国老年学杂志,2013,33(23):5858.
[20] 徐寒松,孔德明,李雪梅,等.通脉糖眼明胶囊对单纯型糖尿病视网膜病变患者闪光视网膜电图及其振荡电位的影响[J].中医杂志,2007,48(6):512.