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Anti-high mobility group box-1 (HMGB1) antibody inhibits hemorrhage-induced brain injury and improved neurological deficits in rats

View Article: PubMed Central - PubMed

ABSTRACT

As one of the most lethal stroke subtypes, intracerebral hemorrhage (ICH) is acknowledged as a serious clinical problem lacking effective treatment. Available evidence from preclinical and clinical studies suggests that inflammatory mechanisms are involved in the progression of ICH-induced secondary brain injury. High mobility group box-1 (HMGB1) is a ubiquitous and abundant nonhistone DNA-binding protein, and is also an important proinflammatory molecule once released into the extracellular space from the nuclei. Here, we show that treatment with neutralizing anti-HMGB1 mAb (1 mg/kg, i.v. twice) remarkably ameliorated ICH-injury induced by local injection of collagenase IV in the striatum of rats. Administration of anti-HMGB1 mAb inhibited the release of HMGB1 into the extracellular space in the peri-hematomal region, reduced serum HMGB1 levels and decreased brain edema by protecting blood-brain barrier integrity, in association with decreased activated microglia and the expression of inflammation-related factors at 24 h after ICH. Consequently, anti-HMGB1 mAb reduced the oxidative stress and improved the behavioral performance of rats. These results strongly indicate that HMGB1 plays a critical role in the development of ICH-induced secondary injury through the amplification of plural inflammatory responses. Intravenous injection of neutralizing anti-HMGB1 mAb has potential as a novel therapeutic strategy for ICH.

No MeSH data available.


Related in: MedlinePlus

Effects of anti-HMGB1 mAb on oxidative stress, cell apoptosis and the impairment of motor functions after ICH.(a) The blood samples were collected from the left ventricle of the heart at 24 h after ICH. Diacron-reactive oxygen metabolites (d-ROM) were measured using serum samples. F(2,20) = 10.953, p < 0.001. (b) Changes in the serum bioantioxidant potentials (BAP) at 24 h after ICH. BAP was measured using serum samples. F(2,20) = 3.833, p = 0.039. (c,d) TUNEL staining to reveal apoptotic cells in the ipsilateral striatum in each group. The scale bar represents 100 μm. F(2,9) = 50.182, p < 0.001. (e) The grip strength test of contralateral forelimb was measured at the indicated times in each rat as described in the Methods section. A hundred % represents the pre-experiment strength. F(7,36) = 9.867, p < 0.001. (f) Contralateral forelimb use in the cylinder tested at the indicated times after ICH. F(7,36) = 4.222, p = 0.002. Results are shown for the sham group (Sham, n = 7, 7, 4 in (a,b,d), respectively), the control IgG-treated group (Con IgG, n = 8, 8, 4, 5, 5 in (a,b,d,e,f), respectively), and anti-HMGB1 mAb-treated group (α-HMGB1, n = 8, 8, 4, 6, 6 to (a,b,d,e,f), respectively). Values represent the means ± SEM. *p < 0.05, **p < 0.01 compared with the sham group. #p < 0.05 compared with the control IgG-treated group.
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f8: Effects of anti-HMGB1 mAb on oxidative stress, cell apoptosis and the impairment of motor functions after ICH.(a) The blood samples were collected from the left ventricle of the heart at 24 h after ICH. Diacron-reactive oxygen metabolites (d-ROM) were measured using serum samples. F(2,20) = 10.953, p < 0.001. (b) Changes in the serum bioantioxidant potentials (BAP) at 24 h after ICH. BAP was measured using serum samples. F(2,20) = 3.833, p = 0.039. (c,d) TUNEL staining to reveal apoptotic cells in the ipsilateral striatum in each group. The scale bar represents 100 μm. F(2,9) = 50.182, p < 0.001. (e) The grip strength test of contralateral forelimb was measured at the indicated times in each rat as described in the Methods section. A hundred % represents the pre-experiment strength. F(7,36) = 9.867, p < 0.001. (f) Contralateral forelimb use in the cylinder tested at the indicated times after ICH. F(7,36) = 4.222, p = 0.002. Results are shown for the sham group (Sham, n = 7, 7, 4 in (a,b,d), respectively), the control IgG-treated group (Con IgG, n = 8, 8, 4, 5, 5 in (a,b,d,e,f), respectively), and anti-HMGB1 mAb-treated group (α-HMGB1, n = 8, 8, 4, 6, 6 to (a,b,d,e,f), respectively). Values represent the means ± SEM. *p < 0.05, **p < 0.01 compared with the sham group. #p < 0.05 compared with the control IgG-treated group.

Mentions: It is well known that ROS are produced during normal oxidative metabolism, but high ROS levels may damage neurons and cause neuronal death. The serum hydroperoxide concentration was determined and the results are shown in Fig. 8a. Twenty-four hours after ICH, the hydroperoxide concentration increased in the control IgG-treated groups. However, the levels in the anti-HMGB1 group remained significantly lower than that in the control group. To evaluate the oxidant/antioxidant balance after ICH, the serum bioantioxidant potency (BAP) was measured (Fig. 8b). The pattern of serum bioantioxidant potency was roughly parallel to that of the ROS. When comparing the two groups, the BAP in the control group was significantly higher than that in the anti-HMGB1-treated group.


Anti-high mobility group box-1 (HMGB1) antibody inhibits hemorrhage-induced brain injury and improved neurological deficits in rats
Effects of anti-HMGB1 mAb on oxidative stress, cell apoptosis and the impairment of motor functions after ICH.(a) The blood samples were collected from the left ventricle of the heart at 24 h after ICH. Diacron-reactive oxygen metabolites (d-ROM) were measured using serum samples. F(2,20) = 10.953, p < 0.001. (b) Changes in the serum bioantioxidant potentials (BAP) at 24 h after ICH. BAP was measured using serum samples. F(2,20) = 3.833, p = 0.039. (c,d) TUNEL staining to reveal apoptotic cells in the ipsilateral striatum in each group. The scale bar represents 100 μm. F(2,9) = 50.182, p < 0.001. (e) The grip strength test of contralateral forelimb was measured at the indicated times in each rat as described in the Methods section. A hundred % represents the pre-experiment strength. F(7,36) = 9.867, p < 0.001. (f) Contralateral forelimb use in the cylinder tested at the indicated times after ICH. F(7,36) = 4.222, p = 0.002. Results are shown for the sham group (Sham, n = 7, 7, 4 in (a,b,d), respectively), the control IgG-treated group (Con IgG, n = 8, 8, 4, 5, 5 in (a,b,d,e,f), respectively), and anti-HMGB1 mAb-treated group (α-HMGB1, n = 8, 8, 4, 6, 6 to (a,b,d,e,f), respectively). Values represent the means ± SEM. *p < 0.05, **p < 0.01 compared with the sham group. #p < 0.05 compared with the control IgG-treated group.
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Related In: Results  -  Collection

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f8: Effects of anti-HMGB1 mAb on oxidative stress, cell apoptosis and the impairment of motor functions after ICH.(a) The blood samples were collected from the left ventricle of the heart at 24 h after ICH. Diacron-reactive oxygen metabolites (d-ROM) were measured using serum samples. F(2,20) = 10.953, p < 0.001. (b) Changes in the serum bioantioxidant potentials (BAP) at 24 h after ICH. BAP was measured using serum samples. F(2,20) = 3.833, p = 0.039. (c,d) TUNEL staining to reveal apoptotic cells in the ipsilateral striatum in each group. The scale bar represents 100 μm. F(2,9) = 50.182, p < 0.001. (e) The grip strength test of contralateral forelimb was measured at the indicated times in each rat as described in the Methods section. A hundred % represents the pre-experiment strength. F(7,36) = 9.867, p < 0.001. (f) Contralateral forelimb use in the cylinder tested at the indicated times after ICH. F(7,36) = 4.222, p = 0.002. Results are shown for the sham group (Sham, n = 7, 7, 4 in (a,b,d), respectively), the control IgG-treated group (Con IgG, n = 8, 8, 4, 5, 5 in (a,b,d,e,f), respectively), and anti-HMGB1 mAb-treated group (α-HMGB1, n = 8, 8, 4, 6, 6 to (a,b,d,e,f), respectively). Values represent the means ± SEM. *p < 0.05, **p < 0.01 compared with the sham group. #p < 0.05 compared with the control IgG-treated group.
Mentions: It is well known that ROS are produced during normal oxidative metabolism, but high ROS levels may damage neurons and cause neuronal death. The serum hydroperoxide concentration was determined and the results are shown in Fig. 8a. Twenty-four hours after ICH, the hydroperoxide concentration increased in the control IgG-treated groups. However, the levels in the anti-HMGB1 group remained significantly lower than that in the control group. To evaluate the oxidant/antioxidant balance after ICH, the serum bioantioxidant potency (BAP) was measured (Fig. 8b). The pattern of serum bioantioxidant potency was roughly parallel to that of the ROS. When comparing the two groups, the BAP in the control group was significantly higher than that in the anti-HMGB1-treated group.

View Article: PubMed Central - PubMed

ABSTRACT

As one of the most lethal stroke subtypes, intracerebral hemorrhage (ICH) is acknowledged as a serious clinical problem lacking effective treatment. Available evidence from preclinical and clinical studies suggests that inflammatory mechanisms are involved in the progression of ICH-induced secondary brain injury. High mobility group box-1 (HMGB1) is a ubiquitous and abundant nonhistone DNA-binding protein, and is also an important proinflammatory molecule once released into the extracellular space from the nuclei. Here, we show that treatment with neutralizing anti-HMGB1 mAb (1&thinsp;mg/kg, i.v. twice) remarkably ameliorated ICH-injury induced by local injection of collagenase IV in the striatum of rats. Administration of anti-HMGB1 mAb inhibited the release of HMGB1 into the extracellular space in the peri-hematomal region, reduced serum HMGB1 levels and decreased brain edema by protecting blood-brain barrier integrity, in association with decreased activated microglia and the expression of inflammation-related factors at 24&thinsp;h after ICH. Consequently, anti-HMGB1 mAb reduced the oxidative stress and improved the behavioral performance of rats. These results strongly indicate that HMGB1 plays a critical role in the development of ICH-induced secondary injury through the amplification of plural inflammatory responses. Intravenous injection of neutralizing anti-HMGB1 mAb has potential as a novel therapeutic strategy for ICH.

No MeSH data available.


Related in: MedlinePlus