中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
2期
373-376
,共4页
刘朋%郭闻亚%赵晓南%吕艳霞%魏淑明%王秀丽
劉朋%郭聞亞%趙曉南%呂豔霞%魏淑明%王秀麗
류붕%곽문아%조효남%려염하%위숙명%왕수려
糖尿病%疼痛%γ-氨基丁酸受体%环磷酸腺苷反应元件结合蛋白%N-甲基-D-天冬氨酸受体
糖尿病%疼痛%γ-氨基丁痠受體%環燐痠腺苷反應元件結閤蛋白%N-甲基-D-天鼕氨痠受體
당뇨병%동통%γ-안기정산수체%배린산선감반응원건결합단백%N-갑기-D-천동안산수체
Diabetic%Pain%γ-aminobutyric acid receptor%Cyclic adenosine monophosphate response element binding protein%N-methyl-D-aspartate receptor
目的 利用γ-氨基丁酸B型受体(GABAR)受体激动剂(巴氯芬)和拮抗剂(CGP55845),探讨激活GABAB受体对糖尿病神经痛大鼠脊髓背角磷酸化环磷酸腺苷反应元件结合蛋白(p-CREB)和N-甲基-D-天冬氨酸受体2B亚基(NMDA-2B、NR2B)表达的影响.方法 62只SD雄性大鼠随机分为两组:正常对照组(C组)和糖尿病神经痛模型组(D组),腹腔分别注射生理盐水或链脲佐菌素(STZ,60 mg/kg).50只大鼠腹腔注射STZ,4周后36只大鼠成功制备成糖尿病神经痛(DNP)模型并鞘内置管,根据鞘内给药(共20μ1)随机分为3组(n=12):DNP对照组(D1组):生理盐水10 μl+生理盐水10μ；巴氯芬组(D2组):生理盐水10 μ1+巴氯芬0.5μg;CGP55845+巴氯芬组(D3组):CGP55845 10 μg+巴氯芬0.5 μg；12只同周龄正常大鼠腹腔注射生理盐水并鞘内置管作为C组,鞘内注射生理盐水10μl+生理盐水10μl.4组大鼠两次鞘内注射间隔15 min,连续4d,每天鞘内注射前、后30 min测定大鼠50％机械缩足阈值(PWT),各时点分别为:T1、T2、T3、T4,最后1次测完后取大鼠脊髓背角,采用分子生物学方法测定p-CREB、环磷酸腺苷反应元件结合蛋白(CREB)和NR2B受体表达变化.结果 与C组比较,D1、D3两组大鼠T1-T4各时点PWT明显降低(P＜0.05),p-CREB和NR2B蛋白表达及NR2B mRNA表达明显增多(P＜0.05)；与D1组比较,D2组大鼠各时点PWT显著升高(P＜0.05)；与D1组p-CREB(0.76 ±0.13)、NR2B(1.28 ±0.14)蛋白表达、NR2B mRNA表达(0.83±0.10)比较,D2组p-CREB (0.45±0.08)和NR2B(0.88 0.13)蛋白表达及NR2B mRNA表达(0.53±0.08)显著降低(P＜0.05).4组间比较,CREB蛋白表达差异无统计学意义(P＞0.05).结论 激活GABAB受体可使糖尿病神经痛大鼠脊髓背角p-CREB、NR2B蛋白表达下调,抑制糖尿病神经痛.
目的 利用γ-氨基丁痠B型受體(GABAR)受體激動劑(巴氯芬)和拮抗劑(CGP55845),探討激活GABAB受體對糖尿病神經痛大鼠脊髓揹角燐痠化環燐痠腺苷反應元件結閤蛋白(p-CREB)和N-甲基-D-天鼕氨痠受體2B亞基(NMDA-2B、NR2B)錶達的影響.方法 62隻SD雄性大鼠隨機分為兩組:正常對照組(C組)和糖尿病神經痛模型組(D組),腹腔分彆註射生理鹽水或鏈脲佐菌素(STZ,60 mg/kg).50隻大鼠腹腔註射STZ,4週後36隻大鼠成功製備成糖尿病神經痛(DNP)模型併鞘內置管,根據鞘內給藥(共20μ1)隨機分為3組(n=12):DNP對照組(D1組):生理鹽水10 μl+生理鹽水10μ；巴氯芬組(D2組):生理鹽水10 μ1+巴氯芬0.5μg;CGP55845+巴氯芬組(D3組):CGP55845 10 μg+巴氯芬0.5 μg；12隻同週齡正常大鼠腹腔註射生理鹽水併鞘內置管作為C組,鞘內註射生理鹽水10μl+生理鹽水10μl.4組大鼠兩次鞘內註射間隔15 min,連續4d,每天鞘內註射前、後30 min測定大鼠50％機械縮足閾值(PWT),各時點分彆為:T1、T2、T3、T4,最後1次測完後取大鼠脊髓揹角,採用分子生物學方法測定p-CREB、環燐痠腺苷反應元件結閤蛋白(CREB)和NR2B受體錶達變化.結果 與C組比較,D1、D3兩組大鼠T1-T4各時點PWT明顯降低(P＜0.05),p-CREB和NR2B蛋白錶達及NR2B mRNA錶達明顯增多(P＜0.05)；與D1組比較,D2組大鼠各時點PWT顯著升高(P＜0.05)；與D1組p-CREB(0.76 ±0.13)、NR2B(1.28 ±0.14)蛋白錶達、NR2B mRNA錶達(0.83±0.10)比較,D2組p-CREB (0.45±0.08)和NR2B(0.88 0.13)蛋白錶達及NR2B mRNA錶達(0.53±0.08)顯著降低(P＜0.05).4組間比較,CREB蛋白錶達差異無統計學意義(P＞0.05).結論 激活GABAB受體可使糖尿病神經痛大鼠脊髓揹角p-CREB、NR2B蛋白錶達下調,抑製糖尿病神經痛.
목적 이용γ-안기정산B형수체(GABAR)수체격동제(파록분)화길항제(CGP55845),탐토격활GABAB수체대당뇨병신경통대서척수배각린산화배린산선감반응원건결합단백(p-CREB)화N-갑기-D-천동안산수체2B아기(NMDA-2B、NR2B)표체적영향.방법 62지SD웅성대서수궤분위량조:정상대조조(C조)화당뇨병신경통모형조(D조),복강분별주사생리염수혹련뇨좌균소(STZ,60 mg/kg).50지대서복강주사STZ,4주후36지대서성공제비성당뇨병신경통(DNP)모형병초내치관,근거초내급약(공20μ1)수궤분위3조(n=12):DNP대조조(D1조):생리염수10 μl+생리염수10μ；파록분조(D2조):생리염수10 μ1+파록분0.5μg;CGP55845+파록분조(D3조):CGP55845 10 μg+파록분0.5 μg；12지동주령정상대서복강주사생리염수병초내치관작위C조,초내주사생리염수10μl+생리염수10μl.4조대서량차초내주사간격15 min,련속4d,매천초내주사전、후30 min측정대서50％궤계축족역치(PWT),각시점분별위:T1、T2、T3、T4,최후1차측완후취대서척수배각,채용분자생물학방법측정p-CREB、배린산선감반응원건결합단백(CREB)화NR2B수체표체변화.결과 여C조비교,D1、D3량조대서T1-T4각시점PWT명현강저(P＜0.05),p-CREB화NR2B단백표체급NR2B mRNA표체명현증다(P＜0.05)；여D1조비교,D2조대서각시점PWT현저승고(P＜0.05)；여D1조p-CREB(0.76 ±0.13)、NR2B(1.28 ±0.14)단백표체、NR2B mRNA표체(0.83±0.10)비교,D2조p-CREB (0.45±0.08)화NR2B(0.88 0.13)단백표체급NR2B mRNA표체(0.53±0.08)현저강저(P＜0.05).4조간비교,CREB단백표체차이무통계학의의(P＞0.05).결론 격활GABAB수체가사당뇨병신경통대서척수배각p-CREB、NR2B단백표체하조,억제당뇨병신경통.
Objective To investigate the effect of activating γ-aminobutyric acid(GABAB) receptors on the expression of phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) and N-methyl-D-aspartate receptor subunit 2B (NR2B) receptors in the spinal dorsal horn in rats with diabetic neuropathic pain (DNP) by using GABAB receptors agonist (baclofen) and atagonist (CGP55845).Methods Sixty-two male SD rats were randomly divided into two groups:normal control group (C group) and DNP nodel group (D group),which were intraperitonealy injected with saline and streptozocin (STZ) respectively.Fifty rats were intraperitonealy injected with STZ (60 mg/kg),and 4weeks later,DNP models were successfully established in 36 rats which were randomly divided into 3 groups (n ＝ 12 each) according to the injected medicines:saline 10 μ1 + saline 10 μl were injected intrathecally in D1 group,saline 10 μl + baclofen 0.5 μg in D2 group,CGP55845 10 μg + baclofen 0.5 μg in D3 group.Saline 10 μl + saline 10 μl were injected intrathecally in 12 normal rats as C group.There was an interval of 15 min between twice intrathecal injections in four groups.The PWT was measured at 30 min before and after intrathecal injection for 4 days,and the time points were as follows:T1,T2,T3and T4.The spinal cord dorsal horns of rats were removed after measurement of PWT for detection of the expression of p-CREB,cyclic adenosine monophosphate response element binding protein (CREB) and NR2B receptor.Results As compared with C group,the expression levels of NR2B and p-CREB protein were significantly increased,while PWT was significantly decreased at each time point (T1-T4) in D1 and D3 groups (P＜0.05).As compared with D1 group [p-CREB protein (0.76 ±0.13),NR2B protein (1.28 ±0.14),and NR2B mRNA (0.83 ± 0.10)],the protein expression levels of NR2B (0.88 ±0.13) and p-CREB (0.45 ± 0.08) and NR2B mRNA expression (0.53 ± 0.08) were significantly decreased,and PWT was significantly increased at each time point (T1-T4) in D2 group (P ＜ 0.05).The expression levels of CREB had no significant diffierence among the four groups (P ＞ 0.05).Conclusion Activation of GABAB receptors could down-regulate the expression of p-CREB and NR2B proteins in the spinal cord dorsal horns of rats with DNP,and alleviate DNP.
