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Ischemic postconditioning decreases cerebral edema and brain blood barrier disruption caused by relief of carotid stenosis in a rat model of cerebral hypoperfusion.

Yang F, Zhang X, Sun Y, Wang B, Zhou C, Luo Y, Ge P - PLoS ONE (2013)

Bottom Line: ELISA was used to analyze the expression of MMP-9, claudin-5 and occludin.The activity and location of MMP-9 was analyzed by gelatin zymography and in situ zymography, respectively.The distribution of tight junction proteins claudin-5 and occludin was observed by immunohistochemistry.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, First Bethune Hospital of Jilin University, Changchun, China.

ABSTRACT

Background and purpose: Complications due to brain edema and breakdown of blood brain barrier are an important factor affecting the treatment effects of patients with severe carotid stenosis. In this study, we investigated the protective effects of ischemic postconditioning on brain edema and disruption of blood brain barrier via establishing rat model of hypoperfusion due to severe carotid stenosis.

Methods: Wistar rat model of hypoperfusion due to severe carotid stenosis was established by binding a stainless microtube to both carotid arteries. Ischemic postconditioning procedure consisted of three cycles of 30 seconds ischemia and 30 seconds reperfusion. Brain edema was evaluated by measuring cerebral water content, and blood brain barrier permeability was assayed by examining cerebral concentration of Evans' Blue (EB) and fluorescein sodium (NaF). ELISA was used to analyze the expression of MMP-9, claudin-5 and occludin. The activity and location of MMP-9 was analyzed by gelatin zymography and in situ zymography, respectively. The distribution of tight junction proteins claudin-5 and occludin was observed by immunohistochemistry.

Results: The increased brain water content and cerebral concentration of EB and NaF were suppressed by administration of ischemic postconditioning prior to relief of carotid stenosis. Zymographic studies showed that MMP-9 was mainly located in the cortex and its activity was significantly improved by relief of carotid stenosis and, but the elevated MMP-9 activity was inhibited markedly by ischemic postconditioning. Immunohistochemistry revealed that ischemic postconditioning improved the discontinuous distribution of claudin-5 and occludin. ELISA detected that the expression of up-regulated MMP-9 and down-regulated claudin-5 and occludin caused by carotid relief were all attenuated by ischemic postconditioning.

Conclusions: Ischemic postconditioning is an effective method to prevent brain edema and improve BBB permeability and could be used during relief of severe carotid stenosis.

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Related in: MedlinePlus

Distribution and expression of tight junction proteins caludin-5 and occludin.A, distribution of caludin-5 and occludin in cerebral microvessels. Immuno-histochemical images showed that claudin-5 and occludin were located sharply and continuously on cerebral microvessels in the sham and carotid stenosis group. At 2 day following relief of carotid stenosis, their distribution became discontinuous, which was reversed partly by ischemic postconditioning. B, ELISA assay of claudin-5 concentration. C, ELISA assay of occludin concentration. The level of caludin-5 and occludin in stenosis relief group were significantly lower that those in the sham and the carotid stenosis group at each scheduled time point. However, ischemic postconditioning maintained their quantity. *: p<0.01, versus carotid stenosis group; #: p<0.01, versus stenosis relief group.
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pone-0057869-g004: Distribution and expression of tight junction proteins caludin-5 and occludin.A, distribution of caludin-5 and occludin in cerebral microvessels. Immuno-histochemical images showed that claudin-5 and occludin were located sharply and continuously on cerebral microvessels in the sham and carotid stenosis group. At 2 day following relief of carotid stenosis, their distribution became discontinuous, which was reversed partly by ischemic postconditioning. B, ELISA assay of claudin-5 concentration. C, ELISA assay of occludin concentration. The level of caludin-5 and occludin in stenosis relief group were significantly lower that those in the sham and the carotid stenosis group at each scheduled time point. However, ischemic postconditioning maintained their quantity. *: p<0.01, versus carotid stenosis group; #: p<0.01, versus stenosis relief group.

Mentions: In this section, we tested the distribution of tight junction proteins claudin-5 and occludin on cerebral microvessels and analyzed their changes in quantity. Immuno-histochemical images showed that claudin-5 and occludin were both located sharply and continuously on cerebral microvessels in the carotid stenosis group (Figure 4 A), but they became discontinuous (not circling the cerebral microvessels) in stenosis relief group. However, ischemic postconditioning reversed partly this alteration caused by stenosis relief. For quantifying the quantitative changes of claudin-5 and occludin, ELISA assay was performed. As shown in figure 4 B and C, in comparison with those in the sham and the carotid stenosis group, the level of claudin-5 and occludin decreased significantly from day 1 to day 3 in the stenosis relief group. Nevertheless, ischemic postconditioning suppressed the reduction in these two proteins at each corresponding time point (P<0.01, versus carotid stenosis group). Considering claudin-5 and occludin were the constituents of BBB, we thought that the protection of ischemic postconditioning on permeability of BBB is via maintaining the quantity of claudin-5 and occludin in BBB.


Ischemic postconditioning decreases cerebral edema and brain blood barrier disruption caused by relief of carotid stenosis in a rat model of cerebral hypoperfusion.

Yang F, Zhang X, Sun Y, Wang B, Zhou C, Luo Y, Ge P - PLoS ONE (2013)

Distribution and expression of tight junction proteins caludin-5 and occludin.A, distribution of caludin-5 and occludin in cerebral microvessels. Immuno-histochemical images showed that claudin-5 and occludin were located sharply and continuously on cerebral microvessels in the sham and carotid stenosis group. At 2 day following relief of carotid stenosis, their distribution became discontinuous, which was reversed partly by ischemic postconditioning. B, ELISA assay of claudin-5 concentration. C, ELISA assay of occludin concentration. The level of caludin-5 and occludin in stenosis relief group were significantly lower that those in the sham and the carotid stenosis group at each scheduled time point. However, ischemic postconditioning maintained their quantity. *: p<0.01, versus carotid stenosis group; #: p<0.01, versus stenosis relief group.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585273&req=5

pone-0057869-g004: Distribution and expression of tight junction proteins caludin-5 and occludin.A, distribution of caludin-5 and occludin in cerebral microvessels. Immuno-histochemical images showed that claudin-5 and occludin were located sharply and continuously on cerebral microvessels in the sham and carotid stenosis group. At 2 day following relief of carotid stenosis, their distribution became discontinuous, which was reversed partly by ischemic postconditioning. B, ELISA assay of claudin-5 concentration. C, ELISA assay of occludin concentration. The level of caludin-5 and occludin in stenosis relief group were significantly lower that those in the sham and the carotid stenosis group at each scheduled time point. However, ischemic postconditioning maintained their quantity. *: p<0.01, versus carotid stenosis group; #: p<0.01, versus stenosis relief group.
Mentions: In this section, we tested the distribution of tight junction proteins claudin-5 and occludin on cerebral microvessels and analyzed their changes in quantity. Immuno-histochemical images showed that claudin-5 and occludin were both located sharply and continuously on cerebral microvessels in the carotid stenosis group (Figure 4 A), but they became discontinuous (not circling the cerebral microvessels) in stenosis relief group. However, ischemic postconditioning reversed partly this alteration caused by stenosis relief. For quantifying the quantitative changes of claudin-5 and occludin, ELISA assay was performed. As shown in figure 4 B and C, in comparison with those in the sham and the carotid stenosis group, the level of claudin-5 and occludin decreased significantly from day 1 to day 3 in the stenosis relief group. Nevertheless, ischemic postconditioning suppressed the reduction in these two proteins at each corresponding time point (P<0.01, versus carotid stenosis group). Considering claudin-5 and occludin were the constituents of BBB, we thought that the protection of ischemic postconditioning on permeability of BBB is via maintaining the quantity of claudin-5 and occludin in BBB.

Bottom Line: ELISA was used to analyze the expression of MMP-9, claudin-5 and occludin.The activity and location of MMP-9 was analyzed by gelatin zymography and in situ zymography, respectively.The distribution of tight junction proteins claudin-5 and occludin was observed by immunohistochemistry.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, First Bethune Hospital of Jilin University, Changchun, China.

ABSTRACT

Background and purpose: Complications due to brain edema and breakdown of blood brain barrier are an important factor affecting the treatment effects of patients with severe carotid stenosis. In this study, we investigated the protective effects of ischemic postconditioning on brain edema and disruption of blood brain barrier via establishing rat model of hypoperfusion due to severe carotid stenosis.

Methods: Wistar rat model of hypoperfusion due to severe carotid stenosis was established by binding a stainless microtube to both carotid arteries. Ischemic postconditioning procedure consisted of three cycles of 30 seconds ischemia and 30 seconds reperfusion. Brain edema was evaluated by measuring cerebral water content, and blood brain barrier permeability was assayed by examining cerebral concentration of Evans' Blue (EB) and fluorescein sodium (NaF). ELISA was used to analyze the expression of MMP-9, claudin-5 and occludin. The activity and location of MMP-9 was analyzed by gelatin zymography and in situ zymography, respectively. The distribution of tight junction proteins claudin-5 and occludin was observed by immunohistochemistry.

Results: The increased brain water content and cerebral concentration of EB and NaF were suppressed by administration of ischemic postconditioning prior to relief of carotid stenosis. Zymographic studies showed that MMP-9 was mainly located in the cortex and its activity was significantly improved by relief of carotid stenosis and, but the elevated MMP-9 activity was inhibited markedly by ischemic postconditioning. Immunohistochemistry revealed that ischemic postconditioning improved the discontinuous distribution of claudin-5 and occludin. ELISA detected that the expression of up-regulated MMP-9 and down-regulated claudin-5 and occludin caused by carotid relief were all attenuated by ischemic postconditioning.

Conclusions: Ischemic postconditioning is an effective method to prevent brain edema and improve BBB permeability and could be used during relief of severe carotid stenosis.

Show MeSH
Related in: MedlinePlus