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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (2): 135-139.doi: 10.3736/jcim20090208

• Original Experimental Research • Previous Articles     Next Articles

Effects of Qingkailing effective components on nuclear factor-kappa B in an ischemia-reperfusion injury model of rat brain microvascular endothelial cells in vitro

YH Gao,YW Xing,ZZ Yuan,LQ Zhu,PT Li,SR Wang   

  • Received:2008-08-04 Accepted:2008-09-30 Online:2009-02-20 Published:2009-02-15

Objective

To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-κB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs.
Methods

Brain MVECs of male rats were digested with trypsin and subcultured, then the content of MVECs was adjusted to 1×105/mL and the MVECs were divided into normal control group, untreated group, hyocholic acid group, taurocholic acid group, baicalin group, jasminoidin group, Pinctada martensii group and nimodipine group, with six holes in each group. Except for the normal control group, the MVECs in the other groups were exposed in oxygen and glucose deprivation (OGD) circumstance in vitro to simulate ischemia-reperfusion injury. Immunocytochemical staining and image analysis system were used to observe the expression of NF-κB protein.
Results

Under a light microscope, the nuclei of MVECs in the normal control group were blank. Staining intensity of NF-κB protein in the nucleus in the untreated group was much deeper than that in the endochylema, with NF-κB shifted to nucleus after activation; a small quantity of NF-κB protein were expressed in the border of nucleus next to endochylema in groups of Qingkailing effective components, and the NF-κB protein expression was weaker than that in the untreated group. With the image analysis, we found that transmittance of nucleus and endochylema in the untreated group was significantly lower than that in the normal control group (P<0.01). Transmittance of nucleus and endochylema in the treated groups was higher than that in the untreated group (P<0.05, P<0.01).
Conclusion

Qingkailing effective components have significant effect in inhibiting NF-κB protein transferring from endochylema to nucleus in vitro.

Table 1

Effects of Qingkailing effective components on specific transparence value of MVEC nucleus vs endochylema (x±s)"

Group n Specific transparence value
(nucleus/endochylema)
Normal control 9 1.014±0.097**
Untreated 9 0.934±0.030
Nimodipine 9 1.003±0.063**
Hyocholic acid 9 1.009±0.028**
Baicalin 9 0.997±0.028*
Jasminoidin 9 1.015±0.016**
Pinctada martensii 9 1.002±0.019*
Taurocholic acid 9 1.015±0.067**
[1] Berti R, Williams AJ, Moffett JR, Hale SL, Velarde LC, Elliott PJ, Yao C, Dave JR, Tortella FC . Quantitative real-time RT-PCR analysis of inflammatory gene expression associated with ischemia-reperfusion brain injury[J]. J Cereb Blood Flow Metab, 2002,22(9):1068-1079
doi: 10.1097/00004647-200209000-00004
[2] Hara H, Fink K, Endres M, Friedlander RM, Gagliardini V, Yuan J, Moskowitz MA . Attenuation of transient focal cerebral ischemic injury in transgenic mice expressing a mutant ICE inhibitory protein[J]. J Cere Blood Flow Metab, 1997,17(4):370-375
doi: 10.1097/00004647-199704000-00002
[3] Gao YH, Yuan ZZ, Niu FL, Zhu LQ, Li PT, Wang SR . Effect of new QingKaiLing injection on adhesion molecules expression of an in vitro ischemia and reperfusion injury model of rat brain microvascular endothelial cell[J]. Zhonghua Zhong Yi Yao Za Zhi, 2008,23(3):197-200
高永红, 袁拯忠, 牛福玲, 朱陵群, 李澎涛, 王硕仁 . 新清开灵注射液对大鼠脑缺血再灌注损伤微血管内皮细胞黏附分子表达的影响[J]. 中华中医药杂志, 2008,23(3):197-200
[4] Lou J, Gasche Y, Zheng L, Critico B, Monso-Hinard C, Juillard P, Morel P, Buurman WA, Grau GE . Differential reactivity of brain microvascular endothelial cells to TNF reflects the genetic susceptibility to cerebral malaria[J]. Eur J Immunol, 1998,28(12):3989-4000
doi: 10.1002/(ISSN)1521-4141
[5] Gao YH, Yuan ZZ, Niu FL, Cui W, Zhu LQ, Wang SR . The protective effect of cholic acid on an in vitro injury model of ischemia-reperfusion in cerebral microvascular endothelial cells[J]. Zhongguo Zhong Yao Za Zhi, 2005,30(12):930-932
高永红, 袁拯忠, 牛福玲, 崔巍, 朱陵群, 王硕仁 . 脑微血管内皮细胞体外模拟缺血再灌注模型的建立及牛胆酸的保护作用[J]. 中国中药杂志, 2005,30(12):930-932
[6] Zhang W, Smith C, Shapiro A, Monette R, Hutchison J, Stanimirovic D . Increased expression of bioactive chemokines in human cerebromicrovascular endothelial cells and astrocytes subjected to simulated ischemia in vitro[J]. J Neuroimmunol, 1999,101(2):148-160
doi: 10.1016/S0165-5728(99)00137-X
[7] Chan PH . Reactive oxygen radicals in signaling and damage in the ischemic brain[J]. J Cereb Blood Flow Metab, 2001,21(1):2-14
doi: 10.1097/00004647-200101000-00002
[8] Sun Z, Andersson R . NF-KappaB activation and inhibition: a review[J]. Shock, 2002,18(2):99-106
doi: 10.1097/00024382-200208000-00001
[9] Wei D, Wan Q . The role of nuclear factor-κB in the pathophysiologic processes of cerebral ischemia[J]. Guo Wai Yi Xue Nao Xue Guan Ji Bing Fen Ce, 2004,12(4):285-288
魏东, 万琪 . 核因子κB在脑缺血病理生理过程中的作用[J]. 国外医学脑血管疾病分册, 2004,12(4):285-288
[10] Goebeler M, Gillitzer R, Kilian K, Utzel K, Brocker EB, Rapp UR, Ludwig S . Multiple signaling pathways regulate NF KappaB-dependent transcription of the monocyte chemoattractant protein-1 gene in primary endothelial cell[J]. Blood, 2001,97(1):46-55
doi: 10.1182/blood.V97.1.46
[11] Brown RC, Mark KS, Egleton RD, Huber JD, Burroughs AR, Davis TP . Protection against hypoxia-induced increase in blood-brain barrier permeability: role of tight junction proteins and NFkappaB[J]. J Cell Sci, 2002,116(4):693-700
[12] Howard EF, Chen Q, Cheng C, Carroll JE, Hess D . NF kappaB is activated and ICAM-1 gene expression is upregulated during reoxygenation of human brain endothelial cells[J]. Neurosci Lett, 1998,248(3):199-203
doi: 10.1016/S0304-3940(98)00239-0
[13] Hess DC, Howard E, Cheng C, Carroll J, Hill WD, Hsu CY . Hypertonic mannitol loading of NF-κB transcription factor decoys in human brain microvascular endothelial cells blocks upregulation of ICAM-1[J]. Stroke, 2000,31(5):1179-1186
doi: 10.1161/01.STR.31.5.1179
[14] Sharp FR, Lu A, Tang Y, Millhorn DE . Multiple molecular penumbras after focal cerebral ischemia[J]. J Cereb Blood Flow Metab, 2000,20(7):1011-1032
doi: 10.1097/00004647-200007000-00001
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