荞麦花叶芦丁对高糖刺激血管损伤的保护作用

doi:10.13481/j.1671-587x.20160109

摘要/Abstract

摘要：

目的: 观察荞麦花叶芦丁(RBFL)对高糖所致血管损伤的保护作用,并探讨其可能机制。方法: 45只SD大鼠随机取出8只作为正常对照组,其余37只空腹一次性腹腔注射链脲佐菌素(STZ,60 mg·kg^-1)建立1型糖尿病动物模型。将造模成功大鼠分为模型组 (n=10)和低剂量(45 mg·kg^-1,n=8)、中剂量(90 mg·kg^-1,n=8)及高剂量(180 mg·kg^-1,n=8)RBFL干预组,灌胃给药,每天1次,连续8周;正常对照组和模型组大鼠灌胃等量生理盐水。给药8周后观察各组大鼠的一般状况,记录血糖和体质量,检测不同浓度乙酰胆碱 (Ach)(1×10^-9~1×10^-5 mol·L^-1)和硝普钠 (SNP)(1×10^-10~1×10^-5 mol·L^-1)诱导的各组大鼠胸主动脉血管舒张率,检测各组大鼠血清中氧自由基(ROS)、一氧化氮(NO)水平和超氧化物歧化酶(SOD)、谷胱甘肽-过氧化物酶(GSH-Px)及总一氧化氮合酶(tNOS)活性。正常大鼠处死并迅速分离胸主动脉,分为正常葡萄糖对照组(11 mmol·L^-1 葡萄糖),高浓度葡萄糖损伤(HG)组(44 mmol·L^-1 葡萄糖),低剂量RBFL(0.2 mg·L^-1)、中剂量RBFL(0.4 mg·L^-1)和高剂量RBFL(0.8 mg·L^-1)处理组,HG+亚硝基左旋精氨酸甲酯(L-NAME)组,HG+L-NAME+RBFL组,HG+亚甲蓝(MB)组,HG+MB+RBFL组(n=6)。检测不同浓度Ach 和SNP诱导的各组大鼠胸主动脉血管舒张率,检测正常葡萄糖对照组、HG和RBFL处理组大鼠胸主动脉组织中SOD和NOS活性。结果: 与正常对照组比较,模型组大鼠体质量下降,血糖升高(P<0.01);与模型组比较,RBFL干预组大鼠体质量升高,血糖降低(P<0.05或P<0.01)。与正常对照组比较,模型组大鼠胸主动脉Ach诱导的血管舒张率降低(P<0.05或P<0.01);与模型组比较,RBFL干预组Ach诱导的血管舒张率均升高(P<0.05或P<0.01),且该作用可被L-NAME和MB所取消;各组大鼠SNP诱导的血管舒张率比较差异无统计学意义(P>0.05)。与正常对照组比较,模型组大鼠血清ROS水平升高(P<0.01),NO水平及SOD和NOS活性降低(P<0.01);与模型组比较, 高剂量RBFL干预组大鼠血清ROS水平降低(P<0.01), NO水平及SOD和NOS活性升高(P<0.01)。各组大鼠血清GSH-Px活性比较差异无统计学意义(P>0.05)。与正常葡萄糖对照组比较,HG组大鼠Ach诱导的胸主动脉血管舒张率降低(P<0.01);与HG组比较,不同剂量RBFL(0.2、0.4和0.8 mg·L^-1)处理组大鼠Ach诱导的血管舒张率均升高(P<0.05或P<0.01),且该作用可被L-NAME和MB所取消;各组大鼠SNP诱导的血管舒张率比较差异无统计学意义(P>0.05)。与正常葡萄糖对照组比较,HG组大鼠血管组织中SOD和tNOS活性下降(P<0.01);与HG组比较,高剂量RBFL 处理组大鼠血管组织中SOD和tNOS活性升高(P<0.01)。结论: RBFL慢性整体给药和急性处理可以有效改善小鼠高糖刺激血管内皮依赖性舒张功能,其保护作用与增强血管SOD活性、减少ROS过量生成及促进血管内皮细胞合成NO有关。

关键词: 荞麦芦丁, 高糖血管损伤, 氧化应激, 血管内皮, 一氧化氮, 血管舒张

Abstract:

Objective: To observe the protective effect of rutin from buckwheat flowers and leaves (RBFL)on the vascular injury induced by high glucose stimulation,and to explore the possible mechanism.Methods: Eight rats were randomly selected from forty-five SD rats and used as control group,the last were used to induce the type Ⅰ diabetes mellitus model with STZ (60 mg·kg^-1,i.p.). The successful model rats were divided into model group (n=10),low dose (45 mg·kg^-1)of RBFL group,moderate dose (90 mg·kg^-1)of RBFL group and high dose (180 mg·kg^-1)of RBFLgroup (n=8). The rats in RBFL groups were gavaged the corresponding doses of RBFL once a day for 8 weeks;the rats in control group and model group were gavaged the same volume saline. The general condition was observed after 8 weeks;the blood glucose and body weight were recorded;the vessel relaxant rates of thoracic aortas induced by acetylcholine (Ach)(10^-9-10^-5 mol·L^-1)and sodium nitroprusside (SNP)(10^-10-10^-5 mol·L^-1)of the rats in various groups were detected;the serum reactive oxygen species (ROS)level,nitric oxide (NO)level,and superoxide dismutase (SOD),glutathione peroxidase (GSH-Px)and total nitric oxide synthase (tNOS)activities of the rats in various groups were detected. The thoracic aortas were separated rapidly from the normal rats and divided into normal glucose group (11 mmol·L^-1 glucose),high glucose(HG)group (44 mmol·L^-1 glucose),low RBFL(0.2 mg·L^-1)group,moderate RBFL(0.4 mg·L^-1)group,high RBFL(0.8 mg·L^-1)group,HG+ NG-nitro-L-arginine methyl ester (L-NAME)group,HG+L-NAME +RBFL group,HG+methylene blue (MB)group,and HG+MB +RBFL group. The vessel relaxant rates of thoracic aortas of the rats in various groups induced by Ach and SNP were detected;the SOD and NOS activities in thoracic aortas of the rats in normal group,HG group and RBFL group were detected.Results: Compared with control group,the body weights of the rats in model group were decreased and the blood glucose was increased (P<0.01);compared with model group,the body weights of the rats in RBFL group were increased and the blood glucose was decreased (P<0.05 or P<0.01);compared with control group,the vessel relaxant rate induced by Ach in model group was decreased (P<0.05 or P<0.01);compared with model group,the vessel relaxant rates induced by Ach in RBFL groups were increased (P<0.05 or P<0.01)and the effect was cancelled by L-NAME and MB,while the SNP-induced vessel relaxant rates had no changes (P>0.05). Compared with control group,the level of serum ROS in model group was increased(P<0.01);the NO level,the SOD and NOS activities were decreased(P<0.01).Compared with model group,the level of serum ROS in high RBFL group was decreased;the NO level,the activities of SOD and tNOS were increased (P<0.01),but the serum GSH-Px activity had no changes (P>0.05). Compared with normal glucose group,the vessel relaxant rate induced by Ach in HG group was decreased (P<0.01);compared with HG group,the vessel relaxant rates induced by Ach in RBFL groups were increased (P<0.05 or P<0.01)and the effect was cancelled by L-NAME and MB,while the SNP-induced vessel relaxant rate had no changes (P>0.05).Compared with normal glucose group,the SOD and NOS activities in HG group were decreased (P<0.01);compared with HG group,the activities of SOD and tNOS in high RBFL group were increased (P<0.01). Conclusion: RBFL could improve the endothelium-dependent vasorelaxation in high glucose injury of the rats and the mechanism may be related to enhancing the SOD activity,reducing the ROS generation and promoting the NO synthesis of endothelium cells.

Key words: rutin from buckwheat flowers and leaves, high glucose vascular injury, oxidative stress, endothelium, nitric oxide, vasorelaxation

中图分类号:

R285.5

张博男, 张静怡, 储金秀, 王树华, 韩淑英. 荞麦花叶芦丁对高糖刺激血管损伤的保护作用[J]. 吉林大学学报(医学版), 2016, 42(01): 40-47.

ZHANG Bonan, ZHANG Jingyi, CHU Jinxiu, WANG Shuhua, HAN Shuying. Protective effect of rutin from buckwheat flowers and leaves on vascular injury induced by high glucose stimulation[J]. Journal of Jilin University Medicine Edition, 2016, 42(01): 40-47.

参考文献

[1] Matsumoto T,Lopes RA,Taguchi K,et al.Linking the beneficial effects of current therapeutic approaches in diabetes to the vascular endothelin system[J].Life Sci,2014,118(2): 129-135.

[2] Giacco F,Brownlee M.Oxidative stress and diabetic complications[J].Circ Res,2010,107(9): 1058-1070.

[3] 张博男,储金秀,韩淑英.荞麦花叶总黄酮的舒张血管作用及其机制[J].中国药理学通报,2010,26(7):952-956.

[4] Zhang BN,Han SY,Wang ZL,ea al.Protective effects of rutin from buckwheat flowers and leaves on diabetic cardiomyopathy[J].Lat Am J Phar,2013,32(5): 674-678.

[5] Canivell S,Gomis R.Diagnosis and classification of autoimmune diabetes mellitus[J].Autoimmun Rev,2014,13(4/5):403-407.

[6] Han SY,Chu JX,Li GM,et al.Effects of rutin from leaves and flowers of buckwheat on angiotensin Ⅱ-induced hypertrophy of cardiac myocytes and proliferation of fibroblasts[J].Lat Am J Phar,2010,29(1): 137-140.

[7] 左风云,董岭,杨英.大鼠1型与2型糖尿病模型的对比研究[J].疾病检测与控制,2015, 9(6):383-384.

[8] Diagnosis and classification of diabetes mellitus[J].Diabetes Care,2013,36(Suppl 1):S67-74.

[9] Vanhoutte PM.Endothelial dysfunction: the first step toward coronary arteriosclerosis[J].Circ J,2009,73(4):595-601.

[10] Batchuluun B,Inoguchi T,Sonoda N,et al.Metformin and liraglutide ameliorate high glucose-induced oxidative stress via inhibition of PKC-NAD(P)H oxidase pathway in human aortic endothelial cells[J].Atherosclerosis,2014,232(1): 156-164.

[11] Toda N,Imamura T,Okamura T.Alteration of nitric oxide-mediated blood flow regulation in diabetes mellitus[J].Pharmacol Ther,2010,127(3): 189-209.

[12] Devi TS,Hosoya K,Terasaki T,et al. The critical role of TX-the NIP in oxidative stress and DNA damage and retinal pericy teapoptosis under high glucose: implications for diabetic reti-nopathy[J].J Exp Cell Res,2013,319(7): 1001-1012.

[13] Dursun B,Dursun E,Suleymanlar G,et al.The effect of hemodialysis on accelerated atherosclerosis in diabetic patients: correlation of carotid artery intima-media thickness with oxidative stress[J].J Diabetes Complications,2009,23(4): 257-264.

[14] 张孝侠,李永华.氧化损伤与糖尿病发病的相关性[J].中国热带医学,2013,13(5):611-613.

[15] 刘江月.梓醇抑制NADPH氧化酶保护2型糖尿病早期血管内皮功能[J].中国中药杂志,2014,39(15):2936-2941.