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Acetylcholinesterase inhibitors reduce neuroinflammation and -degeneration in the cortex and hippocampus of a surgery stress rat model.

Kalb A, von Haefen C, Sifringer M, Tegethoff A, Paeschke N, Kostova M, Feldheiser A, Spies CD - PLoS ONE (2013)

Bottom Line: Surgery accompanied by LPS-treatment led to increased IL-1beta gene and protein upregulation in the cortex and hippocampus but was significantly reduced by physostigmine and neostigmine.Physostigmine and neostigmine significantly decreased the protein expression of IL-1 and TNF-alpha.Along with LPS-treatment, acetylcholinesterase inhibitors reduce the pro-inflammatory response as well as neurodegeneration after surgery in the cortex and hippocampus.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology and Intensive Care Medicine, Campus Charité Mitte and Campus Virchow-Klinikum, Charité, Universitätsmedizin Berlin, Germany.

ABSTRACT
Exogenous stress like tissue damage and pathogen invasion during surgical trauma could lead to a peripheral inflammatory response and induce neuroinflammation, which can result in postoperative cognitive dysfunction (POCD). The cholinergic anti-inflammatory pathway is a neurohumoral mechanism that plays a prominent role by suppressing the inflammatory response. Treatments with acetylcholinesterase inhibitors enhance cholinergic transmission and may therefore act as a potential approach to prevent neuroinflammation. In the presence or absence of acetylcholinesterase inhibitors, adult Wistar rats underwent surgery alone or were additionally treated with lipopolysaccharide (LPS). Physostigmine, which can overcome the blood-brain barrier or neostigmine acting only peripheral, served as acetylcholinesterase inhibitors. The expression of pro- and anti-inflammatory cytokines in the cortex, hippocampus, spleen and plasma was measured after 1 h, 24 h, 3 d and 7 d using Real-Time PCR, western blot analysis or cytometric bead array (CBA). Fluoro-Jade B staining of brain slices was employed to elucidate neurodegeneration. The activity of acetylcholinesterase was estimated using a spectrofluorometric method. Surgery accompanied by LPS-treatment led to increased IL-1beta gene and protein upregulation in the cortex and hippocampus but was significantly reduced by physostigmine and neostigmine. Furthermore, surgery in combination with LPS-treatment caused increased protein expression of IL-1, TNF-alpha and IL-10 in the spleen and plasma. Physostigmine and neostigmine significantly decreased the protein expression of IL-1 and TNF-alpha. Neuronal degeneration and the activity of acetylcholinesterase were elevated after surgery with LPS-treatment and reduced by physostigmine and neostigmine. Along with LPS-treatment, acetylcholinesterase inhibitors reduce the pro-inflammatory response as well as neurodegeneration after surgery in the cortex and hippocampus. This combination may represent a tool to break the pathogenesis of POCD.

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Physostigmine and neostigmine reduce surgery combined with LPS-induced IL-1beta expression in the cortex.IL-1beta expression was measured by quantitative Real-Time PCR in cortex samples extracted 1, 24, and 72 h postintervention (A–C). Also, IL-1beta expression was detected by western blot analysis in cortex samples extracted 1 h postintervention (D–E). Surgery combined with LPS-treatment resulted in an increased gene expression of IL-1beta after 1 and 24 h, which was decreased by physostigmine and neostigmine administration. Results of Real-Time PCR quantification are shown as mean ± SEM (n = 8 per group). Data are normalized to levels of saline treated rats (Control = 100%; bars represent mean ± SEM, n = 8 per group). Blots are representative of a series of three blots. The densitometric data represent the ratio of the IL-1beta band to the corresponding β-actin band density. ***P<0.001 and **P<0.01 represent the difference between LPS and saline treated groups. ##P<0.01 and #P<0.05 represent the difference between LPS and physostigmine or neostigmine treated groups.
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pone-0062679-g001: Physostigmine and neostigmine reduce surgery combined with LPS-induced IL-1beta expression in the cortex.IL-1beta expression was measured by quantitative Real-Time PCR in cortex samples extracted 1, 24, and 72 h postintervention (A–C). Also, IL-1beta expression was detected by western blot analysis in cortex samples extracted 1 h postintervention (D–E). Surgery combined with LPS-treatment resulted in an increased gene expression of IL-1beta after 1 and 24 h, which was decreased by physostigmine and neostigmine administration. Results of Real-Time PCR quantification are shown as mean ± SEM (n = 8 per group). Data are normalized to levels of saline treated rats (Control = 100%; bars represent mean ± SEM, n = 8 per group). Blots are representative of a series of three blots. The densitometric data represent the ratio of the IL-1beta band to the corresponding β-actin band density. ***P<0.001 and **P<0.01 represent the difference between LPS and saline treated groups. ##P<0.01 and #P<0.05 represent the difference between LPS and physostigmine or neostigmine treated groups.

Mentions: As shown in Fig. 1, surgery combined with LPS-treatment led to increase IL-1beta gene expression in the cortex after 1 h (Fig. 1A) and 24 h (Fig. 1B) and was significantly reduced by physostigmine and neostigmine application. After 3 days (Fig. 1C) and 7 days (data not shown) no comparable expression of IL-1beta was detectable. Furthermore, the upregulation of IL-1beta in the cortex was confirmed after 1 h at the protein level by western blot analysis (Fig. 1D–E). Surgery in combination with LPS-treatment caused elevated IL-1beta gene expression in the cortex. The cholinergic activation by physostigmine and neostigmine minimized levels of IL-1beta protein expression. Similar results are shown for the hippocampus (Fig. 2). Surgery and treatment with LPS enhanced IL-1beta gene expression after 1 h (Fig. 2A) and 24 h (Fig. 2B) and was significantly reduced by physostigmine and neostigmine. After 3 days (Fig. 2C) and 7 days (data not shown) no comparable expression of IL-1beta was detectable. Western blot analysis in samples from the hippocampus extracted 24 h postintervention confirmed the upregulation of IL-1beta protein expression after surgery combined with LPS-treatment and its inhibition by physostigmine and neostigmine (Fig. 2D–E). No changes in the expression of the cytokines TNF-alpha and IL-10 were detected in the cortex or in the hippocampus (data not shown).


Acetylcholinesterase inhibitors reduce neuroinflammation and -degeneration in the cortex and hippocampus of a surgery stress rat model.

Kalb A, von Haefen C, Sifringer M, Tegethoff A, Paeschke N, Kostova M, Feldheiser A, Spies CD - PLoS ONE (2013)

Physostigmine and neostigmine reduce surgery combined with LPS-induced IL-1beta expression in the cortex.IL-1beta expression was measured by quantitative Real-Time PCR in cortex samples extracted 1, 24, and 72 h postintervention (A–C). Also, IL-1beta expression was detected by western blot analysis in cortex samples extracted 1 h postintervention (D–E). Surgery combined with LPS-treatment resulted in an increased gene expression of IL-1beta after 1 and 24 h, which was decreased by physostigmine and neostigmine administration. Results of Real-Time PCR quantification are shown as mean ± SEM (n = 8 per group). Data are normalized to levels of saline treated rats (Control = 100%; bars represent mean ± SEM, n = 8 per group). Blots are representative of a series of three blots. The densitometric data represent the ratio of the IL-1beta band to the corresponding β-actin band density. ***P<0.001 and **P<0.01 represent the difference between LPS and saline treated groups. ##P<0.01 and #P<0.05 represent the difference between LPS and physostigmine or neostigmine treated groups.
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Related In: Results  -  Collection

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pone-0062679-g001: Physostigmine and neostigmine reduce surgery combined with LPS-induced IL-1beta expression in the cortex.IL-1beta expression was measured by quantitative Real-Time PCR in cortex samples extracted 1, 24, and 72 h postintervention (A–C). Also, IL-1beta expression was detected by western blot analysis in cortex samples extracted 1 h postintervention (D–E). Surgery combined with LPS-treatment resulted in an increased gene expression of IL-1beta after 1 and 24 h, which was decreased by physostigmine and neostigmine administration. Results of Real-Time PCR quantification are shown as mean ± SEM (n = 8 per group). Data are normalized to levels of saline treated rats (Control = 100%; bars represent mean ± SEM, n = 8 per group). Blots are representative of a series of three blots. The densitometric data represent the ratio of the IL-1beta band to the corresponding β-actin band density. ***P<0.001 and **P<0.01 represent the difference between LPS and saline treated groups. ##P<0.01 and #P<0.05 represent the difference between LPS and physostigmine or neostigmine treated groups.
Mentions: As shown in Fig. 1, surgery combined with LPS-treatment led to increase IL-1beta gene expression in the cortex after 1 h (Fig. 1A) and 24 h (Fig. 1B) and was significantly reduced by physostigmine and neostigmine application. After 3 days (Fig. 1C) and 7 days (data not shown) no comparable expression of IL-1beta was detectable. Furthermore, the upregulation of IL-1beta in the cortex was confirmed after 1 h at the protein level by western blot analysis (Fig. 1D–E). Surgery in combination with LPS-treatment caused elevated IL-1beta gene expression in the cortex. The cholinergic activation by physostigmine and neostigmine minimized levels of IL-1beta protein expression. Similar results are shown for the hippocampus (Fig. 2). Surgery and treatment with LPS enhanced IL-1beta gene expression after 1 h (Fig. 2A) and 24 h (Fig. 2B) and was significantly reduced by physostigmine and neostigmine. After 3 days (Fig. 2C) and 7 days (data not shown) no comparable expression of IL-1beta was detectable. Western blot analysis in samples from the hippocampus extracted 24 h postintervention confirmed the upregulation of IL-1beta protein expression after surgery combined with LPS-treatment and its inhibition by physostigmine and neostigmine (Fig. 2D–E). No changes in the expression of the cytokines TNF-alpha and IL-10 were detected in the cortex or in the hippocampus (data not shown).

Bottom Line: Surgery accompanied by LPS-treatment led to increased IL-1beta gene and protein upregulation in the cortex and hippocampus but was significantly reduced by physostigmine and neostigmine.Physostigmine and neostigmine significantly decreased the protein expression of IL-1 and TNF-alpha.Along with LPS-treatment, acetylcholinesterase inhibitors reduce the pro-inflammatory response as well as neurodegeneration after surgery in the cortex and hippocampus.

View Article: PubMed Central - PubMed

Affiliation: Department of Anesthesiology and Intensive Care Medicine, Campus Charité Mitte and Campus Virchow-Klinikum, Charité, Universitätsmedizin Berlin, Germany.

ABSTRACT
Exogenous stress like tissue damage and pathogen invasion during surgical trauma could lead to a peripheral inflammatory response and induce neuroinflammation, which can result in postoperative cognitive dysfunction (POCD). The cholinergic anti-inflammatory pathway is a neurohumoral mechanism that plays a prominent role by suppressing the inflammatory response. Treatments with acetylcholinesterase inhibitors enhance cholinergic transmission and may therefore act as a potential approach to prevent neuroinflammation. In the presence or absence of acetylcholinesterase inhibitors, adult Wistar rats underwent surgery alone or were additionally treated with lipopolysaccharide (LPS). Physostigmine, which can overcome the blood-brain barrier or neostigmine acting only peripheral, served as acetylcholinesterase inhibitors. The expression of pro- and anti-inflammatory cytokines in the cortex, hippocampus, spleen and plasma was measured after 1 h, 24 h, 3 d and 7 d using Real-Time PCR, western blot analysis or cytometric bead array (CBA). Fluoro-Jade B staining of brain slices was employed to elucidate neurodegeneration. The activity of acetylcholinesterase was estimated using a spectrofluorometric method. Surgery accompanied by LPS-treatment led to increased IL-1beta gene and protein upregulation in the cortex and hippocampus but was significantly reduced by physostigmine and neostigmine. Furthermore, surgery in combination with LPS-treatment caused increased protein expression of IL-1, TNF-alpha and IL-10 in the spleen and plasma. Physostigmine and neostigmine significantly decreased the protein expression of IL-1 and TNF-alpha. Neuronal degeneration and the activity of acetylcholinesterase were elevated after surgery with LPS-treatment and reduced by physostigmine and neostigmine. Along with LPS-treatment, acetylcholinesterase inhibitors reduce the pro-inflammatory response as well as neurodegeneration after surgery in the cortex and hippocampus. This combination may represent a tool to break the pathogenesis of POCD.

Show MeSH
Related in: MedlinePlus