新塔花对ApoE-/-小鼠动脉粥样硬化模型中TLR4/NF-κb信号通路的影响
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摘要
目的观察新塔花水提物抗动脉粥样硬化(AS)的作用机制并对Toll样受体4(TLR4)/核转录因子-κb(NF-κb)信号通路及相关细胞因子的影响并探讨其机制。方法 40只8周龄ApoE小鼠随机分为模型组、阿托伐他汀组[3 mg/(kg·d),溶于0.5 mL蒸馏水]、新塔花水提物高剂量组[32 g/(kg·d)]、新塔花水提物低剂量组[9 g/(kg·d)],每组10只;同品系C57小鼠10只为空白组。所有小鼠给予高脂饲料普通环境饲养。适应性喂养1周后,实验组给予一定剂量药物灌胃,空白组和模型组给予等量蒸馏水灌胃,每周根据小鼠体质量变化调整灌胃剂量。12周后处死小鼠,采集血清及主动脉。胸主动脉用于苏木素伊红(HE)染色;血清用于酶联免疫双抗夹心(ELISA)法和血脂4项检测:三酰甘油(TAG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDLG)、高密度脂蛋白胆固醇(HDL-C);升主动脉用于免疫组织化学(IHC)法检测TLR4、NF-κb蛋白水平。结果血脂检测他汀组、高剂量组、低剂量组具有明显的降脂效果(P <0.01),高剂量组和低剂量组具有一定的降低胆固醇效果(P <0.05);ELISA见模型组细胞黏附因子(ICAM)比空白组明显升高(P <0.01),他汀组、高剂量组、低剂量组低于模型组(P <0.01);模型组基质金属蛋白酶-9(MMP-9)较空白组明显升高(P <0.01),他汀组、高剂量组、低剂量组低于模型组(P <0.01);免疫组化(IHC)结果模型组高于空白组免疫组化分析模型组TLR4、NF-κb表达水平均高于空白组(P <0.01),他汀组、高剂量组和低剂量组TLR4、NF-κb表达水平均低于模型组(P <0.01)。结论新塔花具有预防AS的作用,其机制可能与调控TLR4/NF-κb信号通路有关。
Objective: To observe the Effect of Ziziphora clinopodioides Lam. on the TLR4/NF-κb signaling pathway in Atherosclerosis model Mice,and to explore the effect on the prevention and treatment of atherosclerosis. Methods: Forty ApoE(-/-) mice of 8 weeks' old were divided into the model group,Atorvastatin group [3 mg/(kg·d),dissolved in 0.5 mL distilled water],high-dose group of new tower water extract [32 g/(kg·d)],lowdose group of new tower water extract [9 g/(kg·d)], [supplemental dose data] 10 per group;10 strains of the same line C57 mice were the blank group. All mice were fed a high fat diet in a common environment. After 1 week of adaptive feeding,the experimental group was given a certain dose of drug intragastrically,and the blank group and model group were given equal volume of distilled water. After 12 weeks,the mice were sacrificed and serum and aorta were collected. The thoracic aorta was used for hematoxylin and eosin(HE) staining;serum was used for enzyme-linked immunosorbent assay(ELISA) and four lipid tests(TCHO/TG/HDL-C/LDL-G);ascending aorta was used for the levels of TLR4;NF-κb protein were detected by dose group had significant immunohistochemistry(IHC). Results: Blood lipid test showed that statin group,high-dose group and low-dose group had significant lipid-lowering effect(P < 0.01),while high-dose group and low-dose group had certain cholesterollowering effect(P < 0.05). ELISA showed that ICAM in model group was significantly higher than that in blank group(P < 0.01). ICAM in statin group,high dose group and low dose group was lower than that in model group(P < 0.01). MMP-9 in the model group was significantly higher than that in the blank group(P < 0.01). In statin group,high dose group and low dose group,MMP-9 was lower than that in the model group(P < 0.01). The results of IHC showed that the expression of TLR4 and NF-κb in model group was higher than that in blank group(P <0.01). The expression of TLR4 and NF-κb in statin group,high-dose group and low-dose group were lower than that in the model group(P < 0.01). Conclusion: The mechanism of preventing AS may be related to the regulation of TLR4/NF-κb signaling pathway.
Objective: To observe the Effect of Ziziphora clinopodioides Lam. on the TLR4/NF-κb signaling pathway in Atherosclerosis model Mice,and to explore the effect on the prevention and treatment of atherosclerosis. Methods: Forty ApoE(-/-) mice of 8 weeks' old were divided into the model group,Atorvastatin group [3 mg/(kg·d),dissolved in 0.5 mL distilled water],high-dose group of new tower water extract [32 g/(kg·d)],lowdose group of new tower water extract [9 g/(kg·d)], [supplemental dose data] 10 per group;10 strains of the same line C57 mice were the blank group. All mice were fed a high fat diet in a common environment. After 1 week of adaptive feeding,the experimental group was given a certain dose of drug intragastrically,and the blank group and model group were given equal volume of distilled water. After 12 weeks,the mice were sacrificed and serum and aorta were collected. The thoracic aorta was used for hematoxylin and eosin(HE) staining;serum was used for enzyme-linked immunosorbent assay(ELISA) and four lipid tests(TCHO/TG/HDL-C/LDL-G);ascending aorta was used for the levels of TLR4;NF-κb protein were detected by dose group had significant immunohistochemistry(IHC). Results: Blood lipid test showed that statin group,high-dose group and low-dose group had significant lipid-lowering effect(P < 0.01),while high-dose group and low-dose group had certain cholesterollowering effect(P < 0.05). ELISA showed that ICAM in model group was significantly higher than that in blank group(P < 0.01). ICAM in statin group,high dose group and low dose group was lower than that in model group(P < 0.01). MMP-9 in the model group was significantly higher than that in the blank group(P < 0.01). In statin group,high dose group and low dose group,MMP-9 was lower than that in the model group(P < 0.01). The results of IHC showed that the expression of TLR4 and NF-κb in model group was higher than that in blank group(P <0.01). The expression of TLR4 and NF-κb in statin group,high-dose group and low-dose group were lower than that in the model group(P < 0.01). Conclusion: The mechanism of preventing AS may be related to the regulation of TLR4/NF-κb signaling pathway.
引文
[1]陈伟伟,高润霖,刘力生,等.《中国心血管病报告2016》概要[J].中国循环杂志,2017,32(6):521-530.
[2] BHCk M, Hansson GK. Anti-inflammatory therapies for atherosclerosis[J]. Nature Reviews Cardiology,2015,12(4):199-211.
[3]王阳,米树华,杨关林.动脉粥样硬化机制研究进展[J].中国医药,2017,12(4):619-623.
[4] Maslinska D,Laurekamionowska M,Ma linska S. Toll-like receptors as an innate immunity bridge to neuroinflammation in medulloblastoma[J]. Folia Neuropathologica, 2012, 50(50):375-381.
[5] Prakash P,Kulkarni PP,Lentz SR,et al. Cellular fibronectin containing extra domain A promotes arterial thrombosis in mice through platelet Toll-like receptor 4[J]. Blood,2015,125(20):3164-3172.
[6]刘勇民,沙吾提·伊克木.维吾尔药志(上册)[M].乌鲁木齐:新疆科技卫生出版社,1999:446-451.
[7] Asri Y,Ardestani MF,Rabie M,et al. The effect of environmental factors on growth characteristics, seed germination and essential of Ziziphora clinopodioides Lam[J]. 2016,8(29):91-106.
[8] Shahbazi Y. Chemical compositions, antioxidant and antimicrobial properties of Ziziphora clinopodioides Lam. essential oils collected from different parts of Iran[J]. Journal of Food Science&Technology,2017,13(10):1-13.
[9]吴皓东,李燕,郝宇薇,等.唇香草挥发油急性毒性实验研究[J].新疆医科大学学报,2017,40(9):1133-1135.
[10] Behravan J,Ramezani M,Hassanzadeh MK,et al. Composition,Antimycotic and antibacterial activity of ziziphora clinopodioides Lam. Essential Oil from Iran[J]. Journal of Essential Oil Bearing Plants,2007,10(4):339-345.
[11] Yang Wei Jun,Liu Chong,Gu Zheng-Yi,et al. Protective effects of acacetin isolated from Ziziphora clinopo dioides Lam.(Xintahua)on neonatal rat cardiomyo-cytes[J]. Chinese Medicine,2014,9(1):28-33.
[12]张洪平,周月,李得新.维吾尔药唇香草挥发油抗炎、止咳、祛痰和镇痛的药效学研究[J].中华中医药学刊,2017,36(8):2010-2012.
[13] Shahbazi Y. Chemical compositions, antioxidant and antimicrobial properties of Ziziphora clinopodioides Lam. essential oils collected from different parts of Iran[J]. Journal of Food Science&Technology,2017,13(10):1-13.
[14]沙爱龙,陈红玲,孟庆艳.维药芳香新塔花水提物的抗炎镇痛作用及其急性毒性[J].中国实验方剂学杂志,2012,18(4):202-204.
[15]张选明,安冬青.唇香草化学成分及药理和临床应用研究进展[J].中国中医急症,2018,27(3):548-551.
[16] Shahla SN. Chemical composition and in vitro antibacterial activity of Ziziphora clinopodioides Lam. essential oil against some pathogenic bacteria[J]. African Journal of Microbiology Research,2012,6(7):1504-1508.
[17] Moghadam HD,Sani AM, Sangatash M M. Antifungal activity of essential oil of Ziziphora clinopodioides and the inhibition of aflatoxin B1 production in maize grain[J]. Toxicology&Industrial Health,2016,32(3):493-499.
[2] BHCk M, Hansson GK. Anti-inflammatory therapies for atherosclerosis[J]. Nature Reviews Cardiology,2015,12(4):199-211.
[3]王阳,米树华,杨关林.动脉粥样硬化机制研究进展[J].中国医药,2017,12(4):619-623.
[4] Maslinska D,Laurekamionowska M,Ma linska S. Toll-like receptors as an innate immunity bridge to neuroinflammation in medulloblastoma[J]. Folia Neuropathologica, 2012, 50(50):375-381.
[5] Prakash P,Kulkarni PP,Lentz SR,et al. Cellular fibronectin containing extra domain A promotes arterial thrombosis in mice through platelet Toll-like receptor 4[J]. Blood,2015,125(20):3164-3172.
[6]刘勇民,沙吾提·伊克木.维吾尔药志(上册)[M].乌鲁木齐:新疆科技卫生出版社,1999:446-451.
[7] Asri Y,Ardestani MF,Rabie M,et al. The effect of environmental factors on growth characteristics, seed germination and essential of Ziziphora clinopodioides Lam[J]. 2016,8(29):91-106.
[8] Shahbazi Y. Chemical compositions, antioxidant and antimicrobial properties of Ziziphora clinopodioides Lam. essential oils collected from different parts of Iran[J]. Journal of Food Science&Technology,2017,13(10):1-13.
[9]吴皓东,李燕,郝宇薇,等.唇香草挥发油急性毒性实验研究[J].新疆医科大学学报,2017,40(9):1133-1135.
[10] Behravan J,Ramezani M,Hassanzadeh MK,et al. Composition,Antimycotic and antibacterial activity of ziziphora clinopodioides Lam. Essential Oil from Iran[J]. Journal of Essential Oil Bearing Plants,2007,10(4):339-345.
[11] Yang Wei Jun,Liu Chong,Gu Zheng-Yi,et al. Protective effects of acacetin isolated from Ziziphora clinopo dioides Lam.(Xintahua)on neonatal rat cardiomyo-cytes[J]. Chinese Medicine,2014,9(1):28-33.
[12]张洪平,周月,李得新.维吾尔药唇香草挥发油抗炎、止咳、祛痰和镇痛的药效学研究[J].中华中医药学刊,2017,36(8):2010-2012.
[13] Shahbazi Y. Chemical compositions, antioxidant and antimicrobial properties of Ziziphora clinopodioides Lam. essential oils collected from different parts of Iran[J]. Journal of Food Science&Technology,2017,13(10):1-13.
[14]沙爱龙,陈红玲,孟庆艳.维药芳香新塔花水提物的抗炎镇痛作用及其急性毒性[J].中国实验方剂学杂志,2012,18(4):202-204.
[15]张选明,安冬青.唇香草化学成分及药理和临床应用研究进展[J].中国中医急症,2018,27(3):548-551.
[16] Shahla SN. Chemical composition and in vitro antibacterial activity of Ziziphora clinopodioides Lam. essential oil against some pathogenic bacteria[J]. African Journal of Microbiology Research,2012,6(7):1504-1508.
[17] Moghadam HD,Sani AM, Sangatash M M. Antifungal activity of essential oil of Ziziphora clinopodioides and the inhibition of aflatoxin B1 production in maize grain[J]. Toxicology&Industrial Health,2016,32(3):493-499.