BACKGROUND: Astrocytes are considered to provide nutritional support in the central nervous system. However, recent studies have confirmed that astrocytes also play an important role in chronic pain. OBJECTIVE: To i...BACKGROUND: Astrocytes are considered to provide nutritional support in the central nervous system. However, recent studies have confirmed that astrocytes also play an important role in chronic pain. OBJECTIVE: To investigate the effects of intrathecal injection of fluorocitrate, minocycline or both on astrocyte activation and proliferation in the spinal dorsal horn of compressed dorsal root ganglion in rats. DESIGN, TIME AND SETTING: The neurology randomized controlled animal study was performed at the Jiangsu Institute of Anesthesia Medicine, from September 2006 to April 2007. MATERIALS: A total of 96 male Sprague Dawley rats, aged 6-8 weeks, were selected for this study. Following intrathecal catheterization, 80 rats underwent steel bar insertion into the L4-5 intervertebral foramina to make a stable compression on the L4-5 posterior root ganglion. Thus rat models of ganglion compression were established. Minocycline and fluorocitrate were purchased from Sigma, USA. METHODS: A total of 96 rats were randomly and equally divided into six groups. Rat L4, L5 transverse process and intervertebral foramina were exposed in the sham operation group, but without steel bar insertion. The model group did not receive any manipulations. Rats in the phosphate buffered saline (PBS) group were intrathecally injected with 0.01 mmol/L PBS (20 μL). Rats in the fluorocitrate group were subjected to 1 μmol/L fluorocitrate (20 μL). Rats in the minocycline group were intrathecally injected with 5 g/L minocycline (20 μL). Rats in the minocycline and fluorocitrate group received a mixture (20 μL) of 5 g/L minocycline and 1 μmol/L fluorocitrate. Following model establishment, drugs were administered once a day. MAIN OUTCOME MEASURES: At 7 and 14 days following model induction, glial fibrillary acidic protein expression in the spinal dorsal horn was measured by immunofluorescence microscopy. Six sections with significant glial fibrillary acidic protein -positive expression were obtained to count astrocytes under an inverted microscope. RESULTS: No significant differences in astrocyte count were detected between the fluorocitrate and model groups. Cell bodies were small with a few processes in the fluorocitrate group, compared with the model group. The astrocyte count decreased significantly in the minocycline group and the minocycline and fluorocitrate group compared with the sham operation, model, PBS and fluorocitrate groups (P 〈 0.01). The decrease in astrocyte count was mainly found in layers Ⅲ–Ⅳ of the spinal dorsal horn. Cell body volume was smaller and process numbers were fewer in the minocycline group and the minocycline and fluorocitrate group, compared with the model and PBS groups. CONCLUSION: Fluorocitrate can inhibit astrocyte activation, but does not affect astrocyte proliferation. However, minocycline can inhibit the activation and proliferation of astrocytes.展开更多
BACKGROUND: Inflammatory responses in injured nerves have been recognized as important factors for initially sensitizing nociceptive neurons. Cyclooxygenase (COX) is the rate-limiting enzyme in prostaglandin synthe...BACKGROUND: Inflammatory responses in injured nerves have been recognized as important factors for initially sensitizing nociceptive neurons. Cyclooxygenase (COX) is the rate-limiting enzyme in prostaglandin synthesis, and COX-2 inhibitor is involved in mechanisms of analgesia and anti-inflammation. OBJECTIVE: To investigate the effects of COX-2 inhibitor on thermal and mechanical hyperalgesia, as well as expression of growth associated protein 43 (GAP-43) and nerve growth factor (NGF) in dorsal root ganglion, in a rat model of neuropathic pain due to chronic constriction injury. DESIGN, TIME AND SETTING: A randomized, controlled, comparison study that was performed at the Surgical Department and Pathological Laboratory, Second Affiliated Hospital of Shantou University Medical College from September 2006 to September 2007. MATERIALS: COX-2 inhibitor, Iornoxicam, was purchased from Nycomed Pharmaceutical (Austria); rabbit anti-GAP-43, and rabbit anti-NGF polyclonal antibodies were purchased from Boster, Wuhan, China. METHODS: A total of 50 adult, Wistar rats were randomly assigned to four groups: normal control (n = 5), model (n = 15), normal saline control (n = 15), and Iornoxicam treatment (n =15). With exception of the control group, the sciatic nerve of all rats was loosely ligated to establish a model of chronic constriction injury. The model rats were divided into three subgroups according to varying post-operative survival periods: 3, 7 and 14 days (n = 5), respectively. Rats in the Iornoxicam treatment group were intraperitoneally injected with 1.3 mg/kg lornoxicam every 12 hours throughout the entire experimental procedure. Rats in the normal saline control group were intraperitoneally injected with 1.3 mL/kg saline. MAIN OUTCOME MEASURES: Immunohistochemistry revealed expression of GAP-43 and NGF in the L5 dorsal root ganglions. Mechanical withdrawal threshold and thermal withdrawal latency were used to observe neurological behavioral changes in rats. RESULTS: The relative gray values of GAP-43- and NGF-positive neurons in the model group were remarkably increased compared with the normal control rats (P 〈 0.01), while the relative gray values in the Iomoxicam treatment group were significantly less than the model and normal saline control groups (P 〈 0.01). Mechanical withdrawal threshold and thermal withdrawal latency gradually decreased with increasing injury time in the model, normal saline control, and Iornoxicam treatment groups, and were significantly less than the normal control group (P 〈 0.05). In addition, mechanical withdrawal threshold and thermal withdrawal latency were significantly greater in the Iornoxicam treatment group compared with the model and normal saline control groups (P 〈 0.05). CONCLUSION: Intraperitoneal injection of the COX-2 inhibitor Iornoxicam attenuated mechanical and thermal hyperalgesia induced by sciatic nerve chronic constriction injury and inhibited the increased expression of GAP-43 and NGF.展开更多
Bioassay and GC-MS determination indicated that inhibitors of seeds of Fraxinus mandshurica Rupr were distributed mainly in zones of β-inhibitors (including ABA) and of high and low Rf value. The inhibitory activity ...Bioassay and GC-MS determination indicated that inhibitors of seeds of Fraxinus mandshurica Rupr were distributed mainly in zones of β-inhibitors (including ABA) and of high and low Rf value. The inhibitory activity of outer seed coat of the dormant seed (control treatment) was higher than that of seeds with only the inner coat attached e.g. ABA was 114 and 54ng/g fresh weight respectively. The inhibitory effect of No. 12 zone was even higher than ABA. The inhibitory activity of all the zones decreased after stratification (St) and burying (AO) treatment, especially that of the outer seed coat, whose ABA content decreased to 7(St) and 11(AO) ng / g(fw). However, the decrease in inhibition of root growth lagged bchind'thc decrease in germination. It is concluded that burial is advantageous to regeneration. Inhibitors of the outer seed coat and seeds were mostly destroyed under the adverse conditions of natural dissemination (Di treatment) indicating that Di was not conducive to regeneration. MS analysis revealed that the largest peaks of No. 6, 7 and 12 zones were linoleic acid, olcic acid and 17-C, 18-C, and 20-C long chain fatty acids. Several coumarins were found in No. 6 and 7 zones, and cadincncs, farncsol and scsquitcrpcncs were found in No. 6 and 12 zones having common bioorigins with ABA.展开更多
基金the Social Development Science and Technology Plan Program of Jiangsu Province, No. B2004515
文摘BACKGROUND: Astrocytes are considered to provide nutritional support in the central nervous system. However, recent studies have confirmed that astrocytes also play an important role in chronic pain. OBJECTIVE: To investigate the effects of intrathecal injection of fluorocitrate, minocycline or both on astrocyte activation and proliferation in the spinal dorsal horn of compressed dorsal root ganglion in rats. DESIGN, TIME AND SETTING: The neurology randomized controlled animal study was performed at the Jiangsu Institute of Anesthesia Medicine, from September 2006 to April 2007. MATERIALS: A total of 96 male Sprague Dawley rats, aged 6-8 weeks, were selected for this study. Following intrathecal catheterization, 80 rats underwent steel bar insertion into the L4-5 intervertebral foramina to make a stable compression on the L4-5 posterior root ganglion. Thus rat models of ganglion compression were established. Minocycline and fluorocitrate were purchased from Sigma, USA. METHODS: A total of 96 rats were randomly and equally divided into six groups. Rat L4, L5 transverse process and intervertebral foramina were exposed in the sham operation group, but without steel bar insertion. The model group did not receive any manipulations. Rats in the phosphate buffered saline (PBS) group were intrathecally injected with 0.01 mmol/L PBS (20 μL). Rats in the fluorocitrate group were subjected to 1 μmol/L fluorocitrate (20 μL). Rats in the minocycline group were intrathecally injected with 5 g/L minocycline (20 μL). Rats in the minocycline and fluorocitrate group received a mixture (20 μL) of 5 g/L minocycline and 1 μmol/L fluorocitrate. Following model establishment, drugs were administered once a day. MAIN OUTCOME MEASURES: At 7 and 14 days following model induction, glial fibrillary acidic protein expression in the spinal dorsal horn was measured by immunofluorescence microscopy. Six sections with significant glial fibrillary acidic protein -positive expression were obtained to count astrocytes under an inverted microscope. RESULTS: No significant differences in astrocyte count were detected between the fluorocitrate and model groups. Cell bodies were small with a few processes in the fluorocitrate group, compared with the model group. The astrocyte count decreased significantly in the minocycline group and the minocycline and fluorocitrate group compared with the sham operation, model, PBS and fluorocitrate groups (P 〈 0.01). The decrease in astrocyte count was mainly found in layers Ⅲ–Ⅳ of the spinal dorsal horn. Cell body volume was smaller and process numbers were fewer in the minocycline group and the minocycline and fluorocitrate group, compared with the model and PBS groups. CONCLUSION: Fluorocitrate can inhibit astrocyte activation, but does not affect astrocyte proliferation. However, minocycline can inhibit the activation and proliferation of astrocytes.
基金Supported by:the Scientific Research Program of Xiamen Science and Technology Bureau,No. 3502Z20077074
文摘BACKGROUND: Inflammatory responses in injured nerves have been recognized as important factors for initially sensitizing nociceptive neurons. Cyclooxygenase (COX) is the rate-limiting enzyme in prostaglandin synthesis, and COX-2 inhibitor is involved in mechanisms of analgesia and anti-inflammation. OBJECTIVE: To investigate the effects of COX-2 inhibitor on thermal and mechanical hyperalgesia, as well as expression of growth associated protein 43 (GAP-43) and nerve growth factor (NGF) in dorsal root ganglion, in a rat model of neuropathic pain due to chronic constriction injury. DESIGN, TIME AND SETTING: A randomized, controlled, comparison study that was performed at the Surgical Department and Pathological Laboratory, Second Affiliated Hospital of Shantou University Medical College from September 2006 to September 2007. MATERIALS: COX-2 inhibitor, Iornoxicam, was purchased from Nycomed Pharmaceutical (Austria); rabbit anti-GAP-43, and rabbit anti-NGF polyclonal antibodies were purchased from Boster, Wuhan, China. METHODS: A total of 50 adult, Wistar rats were randomly assigned to four groups: normal control (n = 5), model (n = 15), normal saline control (n = 15), and Iornoxicam treatment (n =15). With exception of the control group, the sciatic nerve of all rats was loosely ligated to establish a model of chronic constriction injury. The model rats were divided into three subgroups according to varying post-operative survival periods: 3, 7 and 14 days (n = 5), respectively. Rats in the Iornoxicam treatment group were intraperitoneally injected with 1.3 mg/kg lornoxicam every 12 hours throughout the entire experimental procedure. Rats in the normal saline control group were intraperitoneally injected with 1.3 mL/kg saline. MAIN OUTCOME MEASURES: Immunohistochemistry revealed expression of GAP-43 and NGF in the L5 dorsal root ganglions. Mechanical withdrawal threshold and thermal withdrawal latency were used to observe neurological behavioral changes in rats. RESULTS: The relative gray values of GAP-43- and NGF-positive neurons in the model group were remarkably increased compared with the normal control rats (P 〈 0.01), while the relative gray values in the Iomoxicam treatment group were significantly less than the model and normal saline control groups (P 〈 0.01). Mechanical withdrawal threshold and thermal withdrawal latency gradually decreased with increasing injury time in the model, normal saline control, and Iornoxicam treatment groups, and were significantly less than the normal control group (P 〈 0.05). In addition, mechanical withdrawal threshold and thermal withdrawal latency were significantly greater in the Iornoxicam treatment group compared with the model and normal saline control groups (P 〈 0.05). CONCLUSION: Intraperitoneal injection of the COX-2 inhibitor Iornoxicam attenuated mechanical and thermal hyperalgesia induced by sciatic nerve chronic constriction injury and inhibited the increased expression of GAP-43 and NGF.
文摘Bioassay and GC-MS determination indicated that inhibitors of seeds of Fraxinus mandshurica Rupr were distributed mainly in zones of β-inhibitors (including ABA) and of high and low Rf value. The inhibitory activity of outer seed coat of the dormant seed (control treatment) was higher than that of seeds with only the inner coat attached e.g. ABA was 114 and 54ng/g fresh weight respectively. The inhibitory effect of No. 12 zone was even higher than ABA. The inhibitory activity of all the zones decreased after stratification (St) and burying (AO) treatment, especially that of the outer seed coat, whose ABA content decreased to 7(St) and 11(AO) ng / g(fw). However, the decrease in inhibition of root growth lagged bchind'thc decrease in germination. It is concluded that burial is advantageous to regeneration. Inhibitors of the outer seed coat and seeds were mostly destroyed under the adverse conditions of natural dissemination (Di treatment) indicating that Di was not conducive to regeneration. MS analysis revealed that the largest peaks of No. 6, 7 and 12 zones were linoleic acid, olcic acid and 17-C, 18-C, and 20-C long chain fatty acids. Several coumarins were found in No. 6 and 7 zones, and cadincncs, farncsol and scsquitcrpcncs were found in No. 6 and 12 zones having common bioorigins with ABA.