黄芩苷通过抑制小胶质细胞活化缓解吗啡耐受

探讨黄芩苷对小胶质细胞活化和慢性耐受的影响并考察其作用机制。应用RT-PCR和ELISA法考察细胞释放炎症因子IL-1β和TNF-α的变化,Western blot法检测细胞中p-p38MAPK和IBA-1蛋白表达变化,热板法检测慢性吗啡耐受小鼠热板痛阈变化。实验结果发现,黄芩苷(1,10,100 μmol/L)剂量依赖性地抑制吗啡活化的BV2细胞释放IL-1β和TNF-α,100 μmol/L黄芩苷明显降低p-p38MAPK水平。鞘内注射黄芩苷(20,40,60 μg)可剂量依赖性地改善慢性吗啡耐受,鞘内注射黄芩苷60 μg可模拟出小胶质细胞抑制剂米诺环素的作用并能显著抑制慢性吗啡耐受小鼠脊髓p-p38MAPK以及IBA-1的表达。本研究表明,黄芩苷可通过抑制p38MAPK信号通路抑制吗啡引起的小胶质细胞活化,进而显著改善慢性吗啡耐受。     

鞘内注射右美托咪定对大鼠吗啡耐受及脊髓炎性反应的影响

目的 探讨鞘内注射右美托咪定对大鼠吗啡耐受及脊髓炎性反应的影响.方法 采用随机数字表法将33只体质量180～200 g雄性SD大鼠,分为3组(n=11):对照组(NS组)、吗啡组(M组)、右美托咪定+吗啡组(Dex组).NS组鞘内注射无菌生理盐水10μL,1次/d,共7 d;M组鞘内注射吗啡15 μg,1次/d,共7 d;Dex组鞘内注射15 μg吗啡和1.5 μg右美托咪定,1次/天,共7d.于吗啡鞘内注射第1、3、5、7天给药前和给药后30 min测定大鼠的热痛阈.Western blot法检测脊髓中小胶质细胞标记物Iba-1、白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和磷酸化p38MAPK(p-p38)的蛋白表达水平.免疫组织化学法检测脊髓Iba-1阳性细胞密度.结果 随着吗啡鞘内注射次数增加,M组和Dex组热水甩尾实验最大镇痛效应百分比(MPE)逐渐下降(P＜0.05).与NS组比较,M组甩尾实验MPE在吗啡注射的第1、3、5、7天较高(P＜0.05);与M组比较,Dex组甩尾实验MPE于吗啡注射的第3、5、7天较高(P＜0.05).与NS组比较,M组大鼠脊髓Iba-1阳性细胞密度及Iba-1、IL-1β、TNF-α和p-p38蛋白表达水平明显升高(P＜0.05);与M组比较,Dex组大鼠脊髓Iba-1阳性细胞密度及Iba-1、IL-1β、TNF-α和p-p38蛋白表达水平明显降低(P＜0.05).结论 鞘内注射右美托咪定可缓解吗啡耐受的形成,其机制可能与抑制脊髓小胶质细胞介导的炎性反应相关.     

宁泌泰胶囊改善慢性前列腺炎引发疼痛的研究

目的 研究宁泌泰(NMT)胶囊改善慢性前列腺炎引发疼痛的作用及潜在机制。方法 将50只雄性C57BL/6小鼠随机分为空白组、模型组、NMT低剂量组、NMT中剂量组、NMT高剂量组,每组10只。造模后低、中、高剂量组分别灌胃给药6、12、18 g·kg^-1NMT溶液,空白组、模型组灌胃等量的生理盐水。采用热辐射甩尾法和热板法进行行为学检测;免疫组化法检测各组前列腺神经胶质纤维酸性蛋白(GFAP)以及肿瘤坏死因子-α(TNF-α)的含量;qPCR法检测前列腺组织中TNF-α、IL-6和IL-1β mRNA的相对表达量;ELISA法检测前列腺组织中IL-6、IL-1β、TNF-α表达水平;Western blot法检测p-ERK/ERK、p-JNK/JNK、p-p38MAPK/p38MAPK蛋白表达水平。结果 与模型组比较,NMT低、中、高3个剂量组小鼠的热板法和甩尾法所测定痛阈值均显著提高(P<0.01,P<0.001),前列腺组织中GFAP和TNF-α表达量明显降低(P<0.05),TNF-α、IL-6和IL-1β的mRNA水平均显著降低(P<...     

p38丝裂原活化蛋白激酶在小鼠肠缺血再灌注肺损伤中的作用

目的探讨p38丝裂原活化蛋白激酶(p38MAPK)在小鼠肠缺血再灌注肺损伤的作用。方法10周龄健康雄性C57BL/6小鼠随机分为假手术组、缺血再灌注组和缺血再灌注+SB239063处理组(SB239063组),SB239063组于术前1h腹腔注入p38MAPK抑制剂SB239063(3mg/kg),另两组注入等量生理盐水。采用夹闭C57BL/6小鼠肠系膜前动脉45min后再灌注6h的方法造成肠缺血再灌注损伤模型。处死小鼠取肺标本,蛋白质印迹法检测肺组织磷酸化p38MAPK蛋白水平,RT-PCR检测肺组织TNF-α和IL-1βmRNA表达,H-E染色观察肺组织病理学改变。结果肠缺血再灌注导致明显肺损伤,肺组织p38MAPK活化明显增加,TNF-α和IL-1β基因表达水平明显升高(与假手术组比较P<0.01);SB239063可抑制肺组织p38MAPK活化,减轻小肠缺血再灌注引起的肺损伤,并下调肺组织TNF-α和IL-1βmRNA表达(与缺血再灌注组比较P<0.05)。结论p38MAPK在小鼠肠缺血再灌注肺损伤中起重要作用,抑制p38MAPK活化可减轻肠缺血再灌注肺损伤。     

祛风通络方对高糖诱导人肾小球系膜细胞炎性反应的影响机制研究

目的 观察祛风通络方对高糖诱导人肾小球系膜细胞炎性反应的影响.方法 体外培养人肾小球系膜细胞株并采用25.0 mmol/L高糖DMEM诱导其增殖,设立正常组、模型组、抑制剂组、祛风通络组,ELISA法测定IL-1β、IL-6、TNF-α、TGF-β1的表达水平,real-time PCR法测定白细胞介素-1β(IL-1β)、IL-6、肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、细胞间黏附分子-1(ICAM-1)、单核细胞趋化蛋白-1(MCP-1)、相对分子量为38×103的促分裂素原活化蛋白激酶(p38 MAPK)mRNA的表达,Western blot法检测ICAM-1、MCP-1、p38 MAPK的表达.结果 与正常组比较,模型组IL-1β、IL-6、TNF-α、TGF-β1水平增加(P<0.05),IL-1β、IL-6、TNF-α、TGF-β1、ICAM-1、MCP-1、p38 MAPK mRNA相对表达水平升高(P<0.05),p38 MAPK、ICAM-1、MCP-1的相对表达水平升高(P<0.05);与模型组比较,祛风通络组IL-1β、IL-6、TNF-α、TGF-β1水平下降(P<0.05),IL-1β、TNF-α、TGF-β1、ICAM-1、MCP-1、p38 MAPK mRNA相对表达水平降低(P<0.05),MCP-1的相对表达水平降低(P<0.05).与抑制剂组比较,祛风通络组p38 MAPK、IL-1β、IL-6、TGF-β1 mRNA相对表达水平,差异有统计学意义(P<0.05)结论祛风通络方可以明显改善高糖诱导人肾小球系膜细胞的炎性反应,减少IL-1β、IL-6、TNF-α、TGF-β1、MCP-1、ICAM-1的表达,这一过程可能是通过p38 MAPK信号通路实现的.     

吗啡耐受大鼠背根神经节磷酸化p38 MAPK的表达

 【目的】 观察鞘内慢性注射吗啡是否激活背根神经节(DRG)神经元,使其p38丝裂原激活的蛋白激酶(p38 MAPK)磷酸化水平发生改变&#65377; 【方法】 通过对成年雄性SD大鼠连续7 d鞘内注射吗啡15 μg而建立吗啡耐受的动物模型,或连续7 d在吗啡注射前30 min预先鞘内注射p38 MAPK抑制剂SB203580 10 μg,用50 ℃热水甩尾法观察吗啡的镇痛效果,并用免疫组织化学法观察DRG 磷酸化p38 MAPK(p-p38 MAPK)的表达变化&#65377;【结果】 慢性鞘内注射吗啡7 d后,吗啡镇痛作用明显下降(P < 0.001),吗啡耐受形成;而SB203580预处理可明显抑制吗啡耐受的形成(P < 0.001);免疫组织化学染色显示脊髓DRG p-p38 MAPK表达明显上调(P < 0.01),且以小&#65380;中型神经元增加为主&#65377; 【结论】 鞘内慢性注射吗啡可激活脊髓DRG小型和中型神经元,使其p38 MAPK磷酸化水平增加,这可能是吗啡耐受的机制之一&#65377;     

P物质刺激下脊髓星形胶质细胞活化中磷酸化P38丝裂素活化蛋白激酶和c-fos的表达及作用

目的 观察P物质(substance P, SP)作用下,脊髓星形胶质细胞中P38丝裂素活化蛋白激酶的磷酸化(P-P38MAPK)和c-fos的表达,及P38MAPK磷酸化对炎性因子分泌的影响.方法 采用体外培养脊髓星形胶质细胞,施加SP刺激,分别采用Western blot和RT-PCR检测P38MAPK磷酸化和c-fos表达的变化,ELISA法检测SB203580阻断P38MAPK磷酸化对炎性因子TNF-α、IL-1β、NO分泌的影响.结果 在SP(10-7mol/L)刺激下,P38MAKP在15、30、45、60 min时均明显磷酸化,与对照组相比有显著性差异 (P＜0.05, P＜0.01),磷酸化高峰在45 min;SP(10-7 mol/L)作用下c-fos表达明显,30 min、1、3、6 h均显著高于对照组 (P＜0.05, P＜0.01),3 h达到高峰;SP(10-7 mol/L)刺激脊髓星形胶质细胞明显分泌炎性因子IL-1β、TNF-α和NO (P＜0.01), SB203580阻断P38MAKP信号通路后TNF-α和NO的分泌下降(P＜0.01),IL-1β的分泌在阻断前后无显著性差异(P＞0.05).结论 SP作用下脊髓星形胶质细胞中P38MAPK和 c-fos信号通路激活,P38MAPK磷酸化参与了SP作用下脊髓星形胶质细胞的活化及炎性因子的分泌,与星形胶质细胞在脊髓中枢对痛觉的调控有关.     

川芎嗪预处理对心肌缺血再灌注损伤大鼠p38MAPK和TNF-α表达的影响

目的:探讨川芎嗪预处理对心肌缺血再灌注损伤的保护作用及机制.方法:30只Wistar大鼠被随机分为3组:假手术组、缺血再灌注损伤(IR)组、川芎嗪预处理组(LI).建立大鼠心肌缺血再灌注损伤模型,应用酶联免疫吸附法,测定心肌TNF-α和IL-6水平;用免疫组化S-P法,检测p38MAPK蛋白的表达.结果:LI组与IR组比较,TNF-α、IL-6均显著降低(P＜0.01),p38MAPK蛋白阳性表达显著减弱(P＜0.01).结论:川芎嗪可降低p38MAPK的活性,抑制TNF-α和IL-6的表达而发挥心肌保护作用.     

槲皮素抑制脊髓小胶质细胞基质金属蛋白酶的表达改善术后疼痛的机制研究

考察槲皮素对足底切口诱发的术后疼痛的影响,并探讨其对脊髓小胶质细胞中基质金属蛋白酶(MMP)-9和MMP-2的抑制作用并考察其作用机制。制备小鼠足底切口术后痛模型,利用Von Frey Hairs检测小鼠机械痛阈值;利用明胶酶谱法检测槲皮素对脊髓水平MMP-9和MMP-2活力的影响;Western blot法检测小鼠脊髓小胶质细胞标记分子钙离子结合蛋白1(IBA-1)以及磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)的变化。结果发现,给予槲皮素(20,40,80 mg/kg,ip)能够显著地抑制足底切口手术引起的机械性痛觉异常以及脊髓小胶质细胞中MMP-9和MMP-2水平的升高;并抑制术后切口引起的脊髓水平IBA-1和p-p38的表达。研究结果表明,槲皮素可能通过抑制脊髓小胶质细胞中MMP-9和MMP-2的活化而改善术后疼痛。     

丙泊酚对小胶质细胞炎症因子的影响及其机制研究

目的 探讨丙泊酚对小胶质细胞炎症因子的影响及其机制。方法 将小胶质BV-2 细胞分为对照组、丙泊酚组、脂多糖（LPS）组、LPS+ 丙泊酚组,对照组细胞加入PBS 液,丙泊酚组细胞加入丙泊酚30μmol/L,LPS 组细胞加入LPS 1μg/ml,LPS+ 丙泊酚组细胞加入丙泊酚30μmol/L+LPS 1μg/ml。采用MTT 比色实验测定细胞活性,采用酶联免疫吸附法测定细胞上清液中白细胞介素1β（IL-1β）、白细胞介素6（IL-6）、肿瘤坏死因子-α（TNF-α）水平,采用逆转录- 聚合酶链反应测定细胞p38MAPK 和TLR4mRNA 的表达,采用Western blot 检测细胞p38MAPK 和TLR4 蛋白表达量。结果 LPS 组和LPS+ 丙泊酚组小胶质细胞活性低于对照组和丙泊酚组（P <0.05）,LPS+ 丙泊酚组小胶质细胞活性高于LPS 组（P <0.05）;LPS 组和LPS+ 丙泊酚组小胶质细胞上清液中IL-1β、IL-6、TNF-α 水平高于对照组和丙泊酚组（P <0.05）,LPS+ 丙泊酚组小胶质细胞上清液中IL-1β、IL-6、TNF-α 水平低于LPS 组（P <0.05）;LPS 组和LPS+ 丙泊酚组小胶质细胞p38MAPK、TLR4 mRNA 和蛋白表达量高于对照组和丙泊酚组（P <0.05）,LPS+ 丙泊酚 组小胶质细胞p38MAPK、TLR4 mRNA 和蛋白表达量低于LPS 组（P <0.05）;丙泊酚组和对照组小胶质细胞各指标比较,差异无统计学意义（P >0.05）。结论 丙泊酚能抑制小胶质细胞过度活化和炎症反应,其机制可能与丙泊酚可下调TLR4-p38MAPK 信号通路有关。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.