左归降糖通脉方含药血清对晚期糖基化终末产物诱导脑微血管内皮细胞损伤的影响
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摘要
目的探讨左归降糖通脉方治疗糖尿病血管并发症的可能作用机制。方法将30只大鼠随机分为空白组、中药组和西药组,每组10只。中药组给予左归降糖通脉方36 g/(kg·d)灌胃,西药组给予尼莫地平片18. 35 mg/(kg·d)及格列齐特缓释片27. 5 mg/(kg·d)灌胃,空白组予以4 ml蒸馏水灌胃,均每日1次,连续5天,制备含药血清。将晚期糖基化终末产物(AGEs)设为50、100、200、400 mg/L 4个浓度组,分别作用12、24、48 h 3个时间点,观察AGEs对大鼠脑微血管内皮细胞(BMECs)形态结构改变的影响来筛选最佳干预浓度和时间。将左归降糖通脉方含药血清分为2. 5%、5%、10%3个浓度组,作用于BMECs 24 h后,测定BMECs增殖率来确定左归降糖通脉方中药含药血清的最佳作用浓度。再将BMECs分为空白组、模型组、中药组、西药组,中药组和西药组分别加入相应含药血清进行干预。检测各组细胞上清液中白细胞介素1β(IL-1β)、细胞间黏附分子1 (ICAM-1)、血管细胞黏附分子1 (VCAM-1)的含量。结果AGEs的最佳干预浓度和时间点为200 mg/L作用24 h,中药含药血清的最佳作用浓度为5%。与空白组比较,模型组上清液中IL-1β、ICAM-1、VCAM-1含量明显升高(P <0. 05);与模型组比较,中药组和西药组IL-1β、ICAM-1、VCAM-1明显下调(P <0. 05)。结论左归降糖通脉方能下调IL-1β、ICAM-1、VCAM-1表达,从而抑制炎症反应,可能是其治疗糖尿病血管并发症的作用机制之一。
Objective To investigate the possible mechanism of Zuogui Jiangtang Tongmai Formulae( ZGJTTMF)( 左归降糖通脉方) in the treatment of diabetic vascular complications. Methods Thirty rats were randomly divided into the blank group,the Chinese medicine group and the western medicine group,with 10 rats in each group. The Chinese medicine group was intragastrically given ZGJTTMF 36 g/( kg ·d),and the western medicine group was given nimodipine tablets 18. 35 mg/( kg·d) and gliclazide sustained-release tablets 27. 5 mg/( kg·d) by gavage. The blank group was intragastrically administered with 4 ml of distilled water. Rats were all given once a day for 5 consecutive days to prepare the drug-containing serum. The advanced glycation end products( AGEs) were set to four concentrations of 50,100,200,and 400 mg/L for 12,24,and 48 h. Effects of AGEs on morphological structural changes of rat brain microvascular endothelial cells( BMECs) were observed to screen for optimal intervention concentrations and times. The drug-containing serum of ZGJTTMF was divided into three concentration groups of2. 5%,5% and 10%. After 24 hours of action on BMECs,the proliferation rate of BMECs was measured to determine the optimal action concentration of serum containing the drug of ZGJTTMF. The BMECs were divided into the blank group,the model group,the Chinese medicine group and the western medicine group. The Chinese medicine group and the western medicine group were respectively added with corresponding drug-containing serum for intervention. The contents of interleukin-1β( IL-1β),intercellular adhesion molecule 1( ICAM-1) and vascular cell adhesion molecule 1( VCAM-1) in the supernatant of each group were detected. Results The optimal concentration and time point of AGEs was 200 mg/L for 24 h,and the optimal concentration of Chinese medicine containing serum was 5%. Compared with the blank group,the contents of IL-1β,ICAM-1 and VCAM-1 in the supernatant of the model group were significantly increased( P < 0. 05). Compared with the model group,IL-1β,ICAM-1 and VCAM-1 in the Chinese medicine group and the western medicine group were significantly down-regulated( P < 0. 05).Conclusion ZGJTTMF can down-regulate the expression of IL-1β,ICAM-1 and VCAM-1,thus inhibiting the inflammatory response,which may be one of the mechanisms of its treatment of diabetic vascular complications.
Objective To investigate the possible mechanism of Zuogui Jiangtang Tongmai Formulae( ZGJTTMF)( 左归降糖通脉方) in the treatment of diabetic vascular complications. Methods Thirty rats were randomly divided into the blank group,the Chinese medicine group and the western medicine group,with 10 rats in each group. The Chinese medicine group was intragastrically given ZGJTTMF 36 g/( kg ·d),and the western medicine group was given nimodipine tablets 18. 35 mg/( kg·d) and gliclazide sustained-release tablets 27. 5 mg/( kg·d) by gavage. The blank group was intragastrically administered with 4 ml of distilled water. Rats were all given once a day for 5 consecutive days to prepare the drug-containing serum. The advanced glycation end products( AGEs) were set to four concentrations of 50,100,200,and 400 mg/L for 12,24,and 48 h. Effects of AGEs on morphological structural changes of rat brain microvascular endothelial cells( BMECs) were observed to screen for optimal intervention concentrations and times. The drug-containing serum of ZGJTTMF was divided into three concentration groups of2. 5%,5% and 10%. After 24 hours of action on BMECs,the proliferation rate of BMECs was measured to determine the optimal action concentration of serum containing the drug of ZGJTTMF. The BMECs were divided into the blank group,the model group,the Chinese medicine group and the western medicine group. The Chinese medicine group and the western medicine group were respectively added with corresponding drug-containing serum for intervention. The contents of interleukin-1β( IL-1β),intercellular adhesion molecule 1( ICAM-1) and vascular cell adhesion molecule 1( VCAM-1) in the supernatant of each group were detected. Results The optimal concentration and time point of AGEs was 200 mg/L for 24 h,and the optimal concentration of Chinese medicine containing serum was 5%. Compared with the blank group,the contents of IL-1β,ICAM-1 and VCAM-1 in the supernatant of the model group were significantly increased( P < 0. 05). Compared with the model group,IL-1β,ICAM-1 and VCAM-1 in the Chinese medicine group and the western medicine group were significantly down-regulated( P < 0. 05).Conclusion ZGJTTMF can down-regulate the expression of IL-1β,ICAM-1 and VCAM-1,thus inhibiting the inflammatory response,which may be one of the mechanisms of its treatment of diabetic vascular complications.
引文
[1]何洪波,祝之明.我国糖尿病合并高血压的流行病学和治疗现状[J].中国科学:生命科学,2018,48(8):855-865.
[2]岳宗相,曾洪长,谢春光,等.糖尿病血管并发症病因学研究进展[J].现代临床医学,2012,38(4):247-250.
[3]YAMAGISHI S,MATSUI T,FUKAMI K. Role of receptor for advanced glycation end products(RAGE)and its ligands in cancer risk[J]. Rejuvenation Res,2015,18(1):48-56.
[4]邓奕辉,喻嵘,陈大舜.降糖通脉方治疗糖尿病合并脑梗死32例临床观察[J].湖南中医学院学报,2004,24(5):40-42.
[5]COOKE JP. Does ADMA cause endothelial dysfunction?[J]. Arterioscler Thromb Vasc Biol,2000,20(9):2032-2037.
[6]贺石林,王键,王净净.中医科研设计与统计学:中医科研设计与统计方法[M].长沙:湖南科学技术出版社,2001:64.
[7]李振宇,谢秀梅,杨天伦,等.前纤维蛋白1在糖基化终末产物介导内皮损伤中的作用[J].中国动脉硬化杂志,2015,23(1):11-16.
[8]李静,张振,姜新魁,等.糖基化终末产物对肠道L细胞株胰高血糖素肽-1分泌水平及L细胞凋亡的影响[J].广东医学,2015,36(7):1013-1016.
[9]廖俞强,邓奕辉,颜佳博,等.滋阴活血解毒方对AGEs诱导VEC损伤黏附分子及凝血相关因子表达的影响[J].湖南中医药大学学报,2017,37(5):485-488.
[10]XUE J,RAY R,SINGER D,et al. The receptor for advanced glycation end products(RAGE)specifically recognizes methylglyoxa L-derived AGEs[J]. Biochemistry,2014,53(20):3327-3335.
[11]BASTA G, LAZZERINI G, MASSARO M, et al.Advanced glycation end products activate endothelium through signaL-transduction receptor RAGE:a mechanism for amplification of inflammatory responses[J]. Circulation,2002,105(7):816-822.
[12]URIBARRI J,DEL CASTILLO MD,DE LA MAZA MP,et al. Dietary advanced glycation end products and their role in health and disease[J]. Adv Nutr,2015,6(4):461-473.
[13]BARZILAY JI,ABRAHAM L,HECKBERT SR,et al.The relation of markers of inflammation to the development of glucose disorders in the elderly:the cardiovascular health study[J]. Diabetes,2001,50(10):2384-2389.
[14]乔爱敏,李乐,刘青.晚期糖基化终产物及其受体RAGE在糖尿病血管并发症中的作用机制[J].中国临床药理学与治疗学,2014,19(11):1306-1311.
[15]高红丽,李虹伟.糖基化终末代谢产物及其受体与动脉粥样硬化的关系及抑制剂的研究进展[J].临床和实验医学杂志,2016,15(6):610-613.
[16]汤运梁,徐积兄.糖基化终末产物与糖尿病慢性并发症研究进展[J].中国老年学杂志,2017,37(4):1012-1015.
[2]岳宗相,曾洪长,谢春光,等.糖尿病血管并发症病因学研究进展[J].现代临床医学,2012,38(4):247-250.
[3]YAMAGISHI S,MATSUI T,FUKAMI K. Role of receptor for advanced glycation end products(RAGE)and its ligands in cancer risk[J]. Rejuvenation Res,2015,18(1):48-56.
[4]邓奕辉,喻嵘,陈大舜.降糖通脉方治疗糖尿病合并脑梗死32例临床观察[J].湖南中医学院学报,2004,24(5):40-42.
[5]COOKE JP. Does ADMA cause endothelial dysfunction?[J]. Arterioscler Thromb Vasc Biol,2000,20(9):2032-2037.
[6]贺石林,王键,王净净.中医科研设计与统计学:中医科研设计与统计方法[M].长沙:湖南科学技术出版社,2001:64.
[7]李振宇,谢秀梅,杨天伦,等.前纤维蛋白1在糖基化终末产物介导内皮损伤中的作用[J].中国动脉硬化杂志,2015,23(1):11-16.
[8]李静,张振,姜新魁,等.糖基化终末产物对肠道L细胞株胰高血糖素肽-1分泌水平及L细胞凋亡的影响[J].广东医学,2015,36(7):1013-1016.
[9]廖俞强,邓奕辉,颜佳博,等.滋阴活血解毒方对AGEs诱导VEC损伤黏附分子及凝血相关因子表达的影响[J].湖南中医药大学学报,2017,37(5):485-488.
[10]XUE J,RAY R,SINGER D,et al. The receptor for advanced glycation end products(RAGE)specifically recognizes methylglyoxa L-derived AGEs[J]. Biochemistry,2014,53(20):3327-3335.
[11]BASTA G, LAZZERINI G, MASSARO M, et al.Advanced glycation end products activate endothelium through signaL-transduction receptor RAGE:a mechanism for amplification of inflammatory responses[J]. Circulation,2002,105(7):816-822.
[12]URIBARRI J,DEL CASTILLO MD,DE LA MAZA MP,et al. Dietary advanced glycation end products and their role in health and disease[J]. Adv Nutr,2015,6(4):461-473.
[13]BARZILAY JI,ABRAHAM L,HECKBERT SR,et al.The relation of markers of inflammation to the development of glucose disorders in the elderly:the cardiovascular health study[J]. Diabetes,2001,50(10):2384-2389.
[14]乔爱敏,李乐,刘青.晚期糖基化终产物及其受体RAGE在糖尿病血管并发症中的作用机制[J].中国临床药理学与治疗学,2014,19(11):1306-1311.
[15]高红丽,李虹伟.糖基化终末代谢产物及其受体与动脉粥样硬化的关系及抑制剂的研究进展[J].临床和实验医学杂志,2016,15(6):610-613.
[16]汤运梁,徐积兄.糖基化终末产物与糖尿病慢性并发症研究进展[J].中国老年学杂志,2017,37(4):1012-1015.