Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by A<italic style="font-style: italic">β</italic><sub>25-35</sub>

Ling WU; Qin ZHENG; Yuan-yuan GUO; Ke-nan ZHANG; Jun LUO; Shuai XIAO; Wen-jing LI; Ming YANG

doi:10.13422/j.cnki.syfjx.20200204

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Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ_25-35

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- Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ_25-35
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 5, Pages: 26-33(2020)
- 作者机构：
  
  江西中医药大学　现代中药制剂教育部重点实验室，南昌　 330004
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200204
  CLC： R2-0; R22; R285.5; R289
- Received：01 July 2019，
  
  Published Online：11 October 2019，
  
  Published：05 March 2020
- 稿件说明：
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Ling WU, Qin ZHENG, Yuan-yuan GUO, et al. Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ_25-35[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 26-33.
DOI：

Ling WU, Qin ZHENG, Yuan-yuan GUO, et al. Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ_25-35[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 26-33. DOI： 10.13422/j.cnki.syfjx.20200204.

摘要

目的：

观察镇心省睡益智方(省睡方)水提液对抗

淀粉样蛋白25-35(A

25-35

)诱导的人脑微血管内皮细胞(HBMEC)损伤的神经保护作用和潜在机制。

方法：

采用A

25-35

诱导的HBMEC细胞损伤作为阿尔茨海默病(AD)细胞模型。本研究分为空白组，A

25-35

组，省睡方水提液低、中、高剂量组(125，250，500 mg·L

-1

)。对其进行相关治疗后，采用噻唑蓝(MTT)比色法确定不同浓度药物及A

25-35

的细胞毒性，采用Hoechst-33258染色观察细胞凋亡情况，采用比色法检测半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性，采用蛋白免疫印迹法(Western blot)检测晚期糖基化终产物受体(RAGE)，低密度脂蛋白受体相关蛋白(LRP1)，葡萄糖转运蛋白1(GLUT1)及葡萄糖转运蛋白3(GLUT3)的表达。

结果：

与空白组比较，A

25-35

组细胞活力显著下降(

0.01)，Hoechst-33258染色观察到明亮的蓝色荧光、染色质固缩、呈致密浓染或碎块状致密浓染，颜色有些发白，凋亡细胞百分比显著增加(

0.01)，Caspase-3活性显著增加(

0.01)，RAGE蛋白表达显著增加(

0.01)，LRP1，GLUT1和GLUT3蛋白表达显著下降(

0.01)；与A

25-35

组比较，省睡方水提液组细胞存活率以剂量依赖性方式显著增加，500 mg·L

-1

省睡方水提液组保护作用比其他组更明显(

0.05)，500 mg·L

-1

省睡方水提液组及显著抑制凋亡细胞的数量(

0.01)，且显著降低Caspase-3活性(

0.01)，125 mg·L

-1

省睡方水提液组RAGE蛋白表达未显著降低，250，500 mg·L

-1

省睡方水提液组RAGE蛋白表达显著下降(

0.01)，而各剂量省睡方水提液组LRP1，GLUT1和GLUT3蛋白表达明显增加(

0.05

0.01)。

结论：

省睡方水提液能够减弱A

25-35

寡聚体诱导的HBMEC细胞毒性，抑制细胞凋亡，降低Caspase-3活性，降低RAGE蛋白的表达，升高LRP1，GLUT1和GLUT3蛋白的表达，从而降低A

在脑内异常聚集和沉积可能是其防治AD的机制。

Abstract

Objective:

To observe the neuroprotective effect and potential mechanism of Zhenxin Shengshui Yizhi Fang(XSF) aqueous extract on human brain microvascular endothelial cells (HBMEC) injury induced by amyloid-

protein(A

)

25-35

Method:

HBMEC cells damage induced by A

25-35

was used as Alzheimer' s disease(AD) cell model. The study included control group

25-35

group

and low

medium and high-dose XSF aqueous extract groups (125

250

500 mg·L

-1

). After treatment

the cytotoxicity of different concentrations of drugs and A

25-35

was determined by methyl thiazolyl tetrazolium(MTT) colorimetry. Apoptosis was observed by Hoechst-33258 staining. The activity of Caspase-3 was detected by colorimetry. Western blot was used to detect the expression levels of the receptor of advanced glycation end products (RAGE) and low-density lipoprotein receptor-related protein (LRP1).

Result:

Compared with the control group

the cell viability of A

25-35

group was significantly decreased (

0.01). Hoechst-33258 staining showed bright blue fluorescence

chromatin condensation

dense staining or fragmentation dense staining

whitening in color

and significant increase of the percentage of apoptotic cells (

0.01). Caspase-3 activity increased significantly (

0.01). Western blot showed that RAGE protein expression increased significantly (

0.01)

while low-density lipoprotein receptor-related protein(LRP1)

glucose transporter 1(GLUT1) and GLUT3 protein expressions decreased significantly (

0.01). Compared with the A

25-35

group

the cell viability of XSF aqueous extract groups was significantly increased in a dose-dependent manner. The XSF aqueous extract had a more significant protective effect of than the other groups (

0.05). The XSF aqueous extract group (500 mg·L

-1

) significantly inhibited the number of apoptotic cells (

0.01)

but significantly reduced the Caspase-3 activity (

0.01). RAGE protein expression was not significantly decreased in XSF aqueous extract group (125 mg·L

-1

)

but significantly decreased in XSF aqueous extract group (250

500 mg·L

-1

0.01)

while LRP1

GLUT1 and GLUT3 protein expression significantly increased (

0.05

0.01) in a dose-dependent manner.

Conclusion:

XSF aqueous extract can attenuate the cytotoxicity of HBMEC induced by A

25-35

oligomer

inhibit apoptosis

decrease caspase-3 activity and RAGE protein expression

increase LRP1

GLUT1 and GLUT3 protein expressions

and reduce the abnormal accumulation and deposition of A

in the brain

which may be its mechanisms for prevention and treatment of AD.

关键词

Keywords

references

刘旭东，王凌志，刘慧慧，等．四君子汤对阿尔茨海默病大鼠行为学及海马神经元能量代谢功能的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 16 ): 1 - 6 .

A KUMAR , A SINGH , EKAVALI . A review on Alzheimer' s disease pathophysiology and its management: an update [J]. Pharmacol Rep ， 2015 ， 67 ( 2 ): 195 - 203 .

韩冉，马涛，张志辰，等．地黄饮子治疗阿尔茨海默病的实验研究进展 [J]．中国实验方剂学杂志， 2018 ， 24 ( 21 ): 124 - 130 .

D PENG , X YUAN , R ZHU . Memantine hydrochloride in the treatment of dementia subtypes [J]． J Clin Neurosci ， 2013 ， 20 ( 11 ): 1482 - 1485 .

L XU , C LI , J YANG . Chemical constituents of Polygala tenuifolia root [J]. J Chin Med Mater ， 2014 ， 37 ( 9 ): 1594 - 1596 .

R K JOSHI . Acorus calamus Linn: phytoconstituents and bactericidal property [J]． World J Microbiol Biotechnol ， 2016 ， 32 ( 10 ): 164 - 170 .

M DENG , L HUANG , Y FANG . Effects of total ginsenosides and volatile oil of acorus tatarinowii co-administration on ability of learning and memory and apoptosis in Alzheimer' s disease mice model induced by D-galactose and aluminium chloride [J]. J Chin Med Mater ， 2015 ， 38 ( 5 ): 1018 - 1026 .

T YABE , H TUCHIDA , H KIYOHARA ， et al . Induction of NGF synthesis in astrocytes by onjisaponins of Polygala tenuifolia，constituents of Kampo (Japanese herbal) medicine，Ninjin-Yoei-To [J]. Phytomedicine (Jena) ， 2003 ， 10 ( 2/3 ): 106 - 114 .

贺密会，刘明，周小江，等．镇心省睡益智方对快动眼睡眠剥夺大鼠促醒和认知功能的影响 [J]．解放军药学学报， 2013 ， 29 ( 5 ): 415 - 419 .

罗俊，张科楠，肖帅，等．镇心省睡益智方及其精油对AD模型小鼠学习记忆的影响 [J]．中国实验方剂学杂志， 2019 ， 25 ( 9 ): 74 - 81 .

W B STINE , K N DAHLGREN , G A KRAFFT ， et al . In vitro characterization of conditions for amyloid-peptide oligomerization and fibrillogenesis [J]. J Biol Chem ， 2003 ， 278 ( 13 ): 11612 - 11622 .

H DU , P LI , J WANG , et al . The interaction of amyloid β and the receptor for advanced glycation endproducts induces matrix metalloproteinase-2 expression in brain endothelial cells [J]. Cell Mol Neurobiol , 2012 , 32 ( 1 ): 141 - 147 .

谭现伟 . A β 1-42 寡聚体制备方法的研究 [D]. 北京：北京交通大学， 2009 .

M A ERICKSON , W A BANKS . Blood brain barrier dysfunction as a cause and consequence of Alzheimer's disease [J]. J Cereb Blood Flow Metab ， 2013 ， 33 ( 49 ): 1500 - 1513 .

C BACHMEIER , M MULLAN , D PARIS . Characterization and use of human brain microvascular endothelial cells to examine β -amyloid exchange in the blood-brain barrier [J]. Cytotechnology ， 2010 ， 62 ( 6 ): 519 - 529 .

R DEANE , R BELL , A SAGARE ， et al . Clearance of amyloid- β peptide across the blood-brain barrier: implication for therapies in alzheimers disease [J]. CNS Neurol Disord Drug Targets ， 2009 ， 8 ( 1 ): 16 - 30 .

M CHRISTOFORIDIS , R SCHOBER , K KROHN ． Genetic-morphologic association study: association between the low density lipoprotein-receptor related protein (LRP) and cerebral amyloid angiopathy [J]． Neuropathol Appl Neurobiol ， 2010 ， 31 ( 1 ): 11 - 19 .

伦琦星，刘炳琳，李军，等．猫耳刺皂苷ilexpernoside C抑制低密度脂蛋白多聚体诱导泡沫细胞形成的活性及机制研究 [J]．中国中药杂志， 2016 ， 41 ( 3 ): 498 - 503 .

B SHACKLETON , F CRAWFORD , C BACHMEIER . Inhibition of ADAM10 promotes the clearance of A β across the BBB by reducing LRP1 ectodomain shedding [J]. Fluids Barriers CNS ， 2016 ， 13 ( 1 ): 78 - 89 .

E A WINKLER , Y NISHIDA , A P SAGARE , et al . GLUT1 reductions exacerbate Alzheimer's disease vasculo-neuronal dysfunction and degeneration [J]. Nat Neurosci , 2015 , 18 ( 4 ): 521 - 530 .

冯慧利，高凯，魏鹏，等 . MicroPET观察姜黄素对9月龄AD小鼠脑葡萄糖代谢的影响 [J]. 中国实验动物学报， 2013 ， 21 ( 4 ): 38 - 41 .

张琦，沈杨，贺文彬．远志、石菖蒲及药对干预AD发病机制的研究概况 [J]．辽宁中医杂志， 2018 ， 45 ( 1 ): 209 - 211 .

郝二伟，邓家刚，侯小涛，等．复方益智颗粒治疗老年性痴呆有效成分的文献研究 [J]．世界中医药， 2016 ， 11 ( 11 ): 2209 - 2212 .

倪杰，房宇，胡燕，等．细叶远志皂苷对阿尔茨海默病大鼠海马低密度脂蛋白受体相关蛋白1水平的影响 [J]．解剖学报， 2016 ， 47 ( 6 ): 744 - 749 .

姜静，邓扬鸥，杨丽． β -细辛醚，丁香酚和远志皂苷对A β 25-35神经细胞毒性的保护作用 [J]．中药药理与临床， 2012 ， 28 ( 5 ): 25 - 28 .

IRIE , W M KEUNG . Rhizoma acori graminei and its active principles protect PC-12 cells from the toxic effect of amyloid- β peptide [J]. Brain Res ， 2003 , 963 ( 1 ): 282 - 289 .

K Y SHIN , B Y WON , C HEO , et al . BT-11 improves stress-induced memory impairments through increment of glucose utilization and total neural cell adhesion molecule levels in rat brains [J]． J Neurosci Res , 2009 , 87 ( 1 ): 260 - 268 .

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⁰

Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ25-35

DOI：10.13422/j.cnki.syfjx.20200204

摘要

Abstract

关键词

Keywords

references

Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ_25-35