2018PubMedRequires access

[High mobility group box 1 promotes apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation by regulating the expression of Bcl-2 and Bax].

M Li, Yaoping Li, Lin Sun, Junlai Song, Cuicui Lv

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Abstract

OBJECTIVE: To investigate the effect of high mobility group protein box 1 (HMGB1) on apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation (OGD/R), and to investigate the possible mechanism by evaluating the expression of apoptosis related protein Bcl-2 and Bax. METHODS: The cerebral cortex astrocytes of neonatal rats were divided into normal group, model group, interference group and control group. Lentivirus vector of rat HMGB1 short hairpin RNA (shRNA) was used to suppress the HMGB1 protein expression in the astrocytes. Then the detection was made after astrocytes were deprived of oxygen and glucose 6 h, reoxygenation for 24 h. The effect of RNA interference was evaluated by Western blotting. The cell survival rate was measured by MTT assay. The apoptosis of astrocytes was determined by TUNEL assay. The expressions of Bcl-2 and Bax were detected by Western blotting. RESULTS: Compared with the normal group, the protein expression of HMGB1 was significantly increased in model group after OGD/R (P<0.001), the astrocytes survival rate was decreased (P<0.001), the number of apoptotic cells labeled with TUNEL was increased (P<0.001), and the ratio of Bcl-2/Bax was decreased (P<0.001). Compared with the model group, RNA interference effectively inhibited the expression of HMGB1 in interference group (P<0.001), the astrocytes survival rate was increased (P<0.001), the number of apoptotic cells labeled with TUNEL was reduced (P<0.01), and the ratio of Bcl-2/Bax was increased (P<0.001). CONCLUSION: The apoptosis of astrocytes can be induced by HMGB1 after OGD/R, and the mechanism may be related to regulating the expression of apoptosis related proteins Bcl-2 and Bax.

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OBJECTIVE: To investigate the effect of high mobility group protein box 1 (HMGB1) on apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation (OGD/R), and to investigate the possible mechanism by evaluating the expression of apoptosis related protein Bcl-2 and Bax. METHODS: The cerebral cortex astrocytes of neonatal rats were divided into normal group, model group, interference group and control group. Lentivirus vector of rat HMGB1 short hairpin RNA (shRNA) was used to suppress the HMGB1 protein expression in the astrocytes. Then the detection was made after astrocytes were deprived of oxygen and glucose 6 h, reoxygenation for 24 h. The effect of RNA interference was evaluated by Western blotting. The cell survival rate was measured by MTT assay. The apoptosis of astrocytes was determined by TUNEL assay. The expressions of Bcl-2 and Bax were detected by Western blotting. RESULTS: Compared with the normal group, the protein expression of HMGB1 was significantly increased in model group after OGD/R (P<0.001), the astrocytes survival rate was decreased (P<0.001), the number of apoptotic cells labeled with TUNEL was increased (P<0.001), and the ratio of Bcl-2/Bax was decreased (P<0.001). Compared with the model group, RNA interference effectively inhibited the expression of HMGB1 in interference group (P<0.001), the astrocytes survival rate was increased (P<0.001), the number of apoptotic cells labeled with TUNEL was reduced (P<0.01), and the ratio of Bcl-2/Bax was increased (P<0.001). CONCLUSION: The apoptosis of astrocytes can be induced by HMGB1 after OGD/R, and the mechanism may be related to regulating the expression of apoptosis related proteins Bcl-2 and Bax.

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Available abstract

OBJECTIVE: To investigate the effect of high mobility group protein box 1 (HMGB1) on apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation (OGD/R), and to investigate the possible mechanism by evaluating the expression of apoptosis related protein Bcl-2 and Bax. METHODS: The cerebral cortex astrocytes of neonatal rats were divided into normal group, model group, interference group and control group. Lentivirus vector of rat HMGB1 short hairpin RNA (shRNA) was used to suppress the HMGB1 protein expression in the astrocytes. Then the detection was made after astrocytes were deprived of oxygen and glucose 6 h, reoxygenation for 24 h. The effect of RNA interference was evaluated by Western blotting. The cell survival rate was measured by MTT assay. The apoptosis of astrocytes was determined by TUNEL assay. The expressions of Bcl-2 and Bax were detected by Western blotting. RESULTS: Compared with the normal group, the protein expression of HMGB1 was significantly increased in model group after OGD/R (P<0.001), the astrocytes survival rate was decreased (P<0.001), the number of apoptotic cells labeled with TUNEL was increased (P<0.001), and the ratio of Bcl-2/Bax was decreased (P<0.001). Compared with the model group, RNA interference effectively inhibited the expression of HMGB1 in interference group (P<0.001), the astrocytes survival rate was increased (P<0.001), the number of apoptotic cells labeled with TUNEL was reduced (P<0.01), and the ratio of Bcl-2/Bax was increased (P<0.001). CONCLUSION: The apoptosis of astrocytes can be induced by HMGB1 after OGD/R, and the mechanism may be related to regulating the expression of apoptosis related proteins Bcl-2 and Bax.

Key concepts: TUNEL assay, Apoptosis, Small hairpin RNA, Blot, HMGB1, Molecular biology, RNA interference, Astrocyte

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[High mobility group box 1 promotes apoptosis of astrocytes after oxygen glucose deprivation/reoxygenation by regulating the expression of Bcl-2 and Bax]. — Research Paper | ScholarLens