电针胃经穴后胃溃疡大鼠胃黏膜细胞蛋白激酶C活性的变化

目的：观察电针胃经（穴）对应激性胃溃疡大鼠胃黏膜细胞蛋白激酶C活性的影响。 方法：实验于2005—07／12在湖南中医药大学针灸基础实验室完成。选择健康SD大鼠60只，随机数字表法分为5组，溃疡血清组（12只）、胃经血清组（12只，取穴：四白、梁门、足三里）和胆经血清组（12只，取穴：阳白、日月、阳陵泉）、胃经血清＋PD153035组（12只）和胆经血清＋PD153035组（12只）。采用水浸束缚法制作胃溃疡动物模型。电针刺激用G6805型电针仪，疏密波。电针频率疏波4Hz。密波20Hz。强度以肌肉或针柄微颤为度，电针时间30min。利用链霉蛋白酶消化法分离胃黏膜细胞，分别用表皮生长因子受体抑制剂PD153035和血清孵育胃黏膜细胞，应用同位素掺入法检测蛋白激酶C活性。 结果：纳入动物60只，均进入结果分析。溃疡血清组大鼠胃黏膜细胞蛋白激酶C有少量表达[（29．16&;#177;30．94）nkat／g]；胃经血清组和胆经血清组大鼠胃黏膜细胞蛋白激酶C呈现较强上增性表达，其中胃经血清组大鼠上增性表达特别明显，两组相比差异有显著性意义[分别为（114．92&;#177;65．61），（36．47&;#177;27.54）nkat／g，P〈0．011；胃经血清＋PD153035组和胆经血清＋PD153035组大鼠胃黏膜细胞蛋白激酶C可见微弱表达[分别为（17.39&;#177;23．39），（23．44&;#177;14．42）nkat／g]，胃经血清组与胃经血清＋PD153035组比较差异有显著性意义（P〈0．01）。 结论：针刺能刺激大鼠胃黏膜细胞信号转导通路中蛋白激酶C的活化，并且存在经脉-脏腑的特异性联系。     

电针胃经穴后胃溃疡大鼠胃黏膜细胞蛋白激酶C活性的变化

目的:观察电针胃经(穴)对应激性胃溃疡大鼠胃黏膜细胞蛋白激酶C活性的影响。方法:实验于2005-07/12在湖南中医药大学针灸基础实验室完成。选择健康SD大鼠60只,随机数字表法分为5组,溃疡血清组穴12只雪、胃经血清组穴12只,取穴:四白、梁门、足三里雪和胆经血清组穴12只,取穴:阳白、日月、阳陵泉雪、胃经血清 PD153035组穴12只雪和胆经血清 PD153035组穴12只雪。采用水浸束缚法制作胃溃疡动物模型。电针刺激用G6805型电针仪,疏密波,电针频率疏波4Hz,密波20Hz,强度以肌肉或针柄微颤为度,电针时间30min。利用链霉蛋白酶消化法分离胃黏膜细胞,分别用表皮生长因子受体抑制剂PD153035和血清孵育胃黏膜细胞,应用同位素掺入法检测蛋白激酶C活性。结果:纳入动物60只,均进入结果分析。溃疡血清组大鼠胃黏膜细胞蛋白激酶C有少量表达眼(29.16±30.94)nkat/g演;胃经血清组和胆经血清组大鼠胃黏膜细胞蛋白激酶C呈现较强上增性表达,其中胃经血清组大鼠上增性表达特别明显,两组相比差异有显著性意义眼分别为(114.92±65.61),(36.47±27.54)nkat/g,P<0.01演;胃经血清 PD153035组和胆经血清 PD153035组大鼠胃黏膜细胞蛋白激酶C可见微弱表达眼分别为(17.39±23.39),(23.44±14.42)nkat/g演,胃经血清组与胃经血清 PD153035组比较差异有显著性意义穴P<0.01雪。结论:针刺能刺激大鼠胃黏膜细胞信号转导通路中蛋白激酶C的活化,并且存在经脉-脏腑的特异性联系。     

电针大鼠胃经后血清对其胃黏膜细胞表皮生长因子受体表达的影响及浓度效应

目的：观察不同稀释度电针胃经后血清对大鼠胃黏膜细胞表皮生长因子受体表达的影响及其浓度效应相关性。方法：实验于2005-07／08在湖南中医药大学针灸基础实验室完成。①选择健康SD大鼠48只，随机分为1／2稀释度血清组、1／5稀释度血清组、1／10稀释度血清组和1／20稀释度血清组，每组12只。②随机选4只大鼠作为血清供体，采用华兴邦动物穴位谱并结合模拟人体经穴法进行取穴，足三里：膝关节后外侧，在腓骨小头下约5mm处。梁门：腹正中线与乳头线之间的中线上，脐上4寸。四白：眶下缘正中。电针刺激用G6805型电针仪，疏密波，电针频率疏波4Hz，密波20Hz，强度以肌肉微颤为度，电针时间30min。将针刺后大鼠行颈动脉取血，离心后吸取血清，将同组4只大鼠血清混合。③另8只大鼠不进行干预。利用链霉蛋白酶消化法分离大鼠胃黏膜细胞。④各组分别用1／2，1／5，1／10，1／20稀释度的血清孵育胃黏膜细胞，采用免疫细胞化学法检测胃黏膜细胞表皮生长因子受体表达水平。结果：纳入动物48只，均进入结果分析。1／2，1／5，1／10和1／20稀释度血清组大鼠胃黏膜细胞表皮生长因子受体均有较强表达，其中1／10稀释度血清组大鼠胃黏膜细胞表皮生长因子受体表达的平均灰度值和面积百分比与1／2，1／5，1／20稀释度血清组比较差异有显著性[平均灰度值：145．43&;#177;5．28，166．35&;#177;9．62，161．38&;#177;6．11，156．53&;#177;4．15；面积百分比：（21．62&;#177;2．08）％，（15．85&;#177;1．35）％，（18．07&;#177;1．28）％，（19．60&;#177;1．10）％，P〈0．05-0．01]。结论：电针胃经后血清可增强胃黏膜细胞表皮生长因子受体的表达，并且存在一定的浓度-效应相关性。     

电针夹背穴在佐剂性关节炎大鼠镇痛过程中磷酸化p38丝裂原活化蛋白激酶电针的变化及作用

目的：观察电针夹脊穴对佐剂性关节炎大鼠背根神经节(dorsal root ganglion，DRG)内磷酸化p38MAPK表达的影响，从信号转导的角度研究电针镇痛的机制。方法：以完全福氏佐剂(CFA)佐剂性关节炎大鼠为疼痛模型，采用免疫组化技术，分别检测空白对照组、模型组、及电针治疗组SD大鼠DGG内磷酸化p38丝裂原活化蛋白激酶(mitogen-activated protein kinase，MAPK)表达特征。结果：与空白对照组[(7．65&;#177;0．72)s]相比，造模后24，48h及7d后，模型组大鼠痛阚下降分别为(6．67&;#177;0．59)，(6．53&;#177;0．73)，(6．64&;#177;0．76)s，同时DRG内p38MAPK磷酸化明显增多(P&;lt;0．05)，尤以48h后增加明显；电针治疗组大鼠在电针夹脊穴治疗后，24，48h及7d后痛阈检测发现其痛阚回升，分别为(7．3&;#177;0．79)，(7．42&;#177;0．93)，和(7．44&;#177;0．89)s，DRG内p38MAPK磷酸化水平下降(P&;lt;0．05)。结论：p38MAPK可能为电针镇痛中一个重要的信号转导分子，其磷酸化在疼痛及电针镇痛过程中其重要作用。     

电针大鼠胃经后血清对其胃黏膜细胞表皮生长因子受体表达的影响及浓度效应

目的:观察不同稀释度电针胃经后血清对大鼠胃黏膜细胞表皮生长因子受体表达的影响及其浓度效应相关性。方法:实验于2005-07/08在湖南中医药大学针灸基础实验室完成。①选择健康SD大鼠48只,随机分为1/2稀释度血清组、1/5稀释度血清组、1/10稀释度血清组和1/20稀释度血清组,每组12只。②随机选4只大鼠作为血清供体,采用华兴邦动物(位谱并结合模拟人体经(法进行取(,足三里:膝关节后外侧,在腓骨小头下约5mm处。梁门:腹正中线与乳头线之间的中线上,脐上4寸。四白:眶下缘正中。电针刺激用G6805型电针仪,疏密波,电针频率疏波4Hz,密波20Hz,强度以肌肉微颤为度,电针时间30min。将针刺后大鼠行颈动脉取血,离心后吸取血清,将同组4只大鼠血清混合。③另8只大鼠不进行干预。利用链霉蛋白酶消化法分离大鼠胃黏膜细胞。④各组分别用1/2,1/5,1/10,1/20稀释度的血清孵育胃黏膜细胞,采用免疫细胞化学法检测胃黏膜细胞表皮生长因子受体表达水平。结果:纳入动物48只,均进入结果分析。1/2,1/5,1/10和1/20稀释度血清组大鼠胃黏膜细胞表皮生长因子受体均有较强表达,其中1/10稀释度血清组大鼠胃黏膜细胞表皮生长因子受体表达的平均灰度值和面积百分比与1/2,1/5,1/20稀释度血清组比较差异有显著性眼平均灰度值:145.43±5.28,166.35±9.62,161.38±6.11,156.53±4.15;面积百分比:(21.62±2.08)%,(15.85±1.35)%,(18.07±1.28)%,(19.60±1.10)%,P<0.05～0.01演。结论:电针胃经后血清可增强胃黏膜细胞表皮生长因子受体的表达,并且存在一定的浓度-效应相关性。     

电针对阿尔茨海默病大鼠学习记忆能力及前额叶P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化通路的影响

目的 观察电针百会、肾俞两穴对阿尔茨海默病(AD)模型大鼠学习记忆能力及前额叶皮质(PFC) P35/P25-周期蛋白依赖性激酶5 (CDK5)-Tau蛋白磷酸化信号通路的影响,以探讨电针治疗AD的相关机制。方法 雄性成年Sprague-Dawley大鼠随机分为正常对照组、假手术组、模型组和电针组,每组6只。后两组双侧海马注射Aβ_25-35,假手术组注射等量生理盐水。造模后次日,电针组电针百会、肾俞,留针15 min,每天1次,共10 d。治疗后,行Morris水迷宫实验,免疫组织化学染色和Western blotting检测各组PFC P35/P25-CDK5-Tau蛋白磷酸化相关蛋白的表达情况。结果 与正常对照组和假手术组比较,模型组的逃避潜伏期和搜索路径均增加(P < 0.05),穿越平台次数减少( P < 0.05);与模型组比较,电针组逃避潜伏期和搜索路径均缩短( P < 0.05),穿越平台次数增加( P < 0.05)。模型组P35/P25和CDK5阳性表达明显高于正常对照组和假手术组( P < 0.01),电针组显著低于模型组( P < 0.001)。模型组P35/P25、CDK5、Tau[pS199]、Tau[pS202]相对表达量高于正常对照组和假手术组( P < 0.05),电针组上述蛋白的表达低于模型组( P < 0.05)。 结论 电针刺激能有效改善AD大鼠的学习记忆和空间探索能力,可能通过影响大鼠PFC的P35/P25-CDK5-Tau蛋白磷酸化信号通路,延缓AD的发生与发展。     

应激后大鼠下丘脑磷酸化p38MAPK的变化及电针足三里穴的调节作用

目的 观察应激对大鼠下丘脑磷酸化p38MAPK的影响及电针足三里穴的调节作用。方法将28只Wistar大鼠随机分为对照组、束缚应激组和束缚应激 电针组，后2组再根据束缚解除后及电针足三里穴位治疗后不同时间点分别分为3个亚组。电针治疗采用频率为5～12Hz的疏密波，治疗总时间30min。于相应时间点处死各组大鼠，采用Western印迹杂交法观察其下丘脑内磷酸化p38MAPK的变化。结果束缚应激组大鼠下丘脑内p38MAPK磷酸化水平明显高于对照组，且在束缚应激解除后3h仍保持在较高水平；应激大鼠经电针足三里穴处理后，下丘脑内磷酸化p38MAPK与束缚应激组相比明显回落，这种调节作用可持续到电针治疗结束后3h。结论束缚应激能引起下丘脑应激信号转导系统中p38MAPK的磷酸化，这可能是应激活动过程中的重要事件之一。电针足三里穴对下丘脑内磷酸化p38MAPK的调节作用可能与下丘脑-垂体-肾上腺轴的活动有关。     

电针对大鼠急性骨骼肌损伤修复中生长相关蛋白和聚集蛋白表达的影响

目的:通过观察电针治疗对急性骨骼肌损伤中生长相关蛋白(GAP-43)及聚集蛋白(agrin)表达的影响,探讨电针促进受损骨骼肌功能恢复的作用机制。方法:将32只SD大鼠随机分为正常组(A组,n=4)、模型组(B组,n=4)、自然恢复组(C组,n=12)、电针组(D组,n=12),A组为空白对照不做任何处理,其余各组使用自制重物打击器建立骨骼肌急性钝挫伤模型,C组不做电针处理自然恢复,D组于造模后48h开始电针干预,每日1次,每次15min。于造模后24h处死B组取材观察造模是否成功,C组及D组于建模后第7、14、21天三个时间点同时取材使用HE染色观察组织形态变化,Western-bolt检测GAP-43及agrin表达情况。结果:HE染色显示:与A组比较,各组肌细胞大量溶解、肌纤维排列紊乱及炎性细胞浸润。D组与C组比较,肌卫星细胞增殖较快,新生肌纤维明显增多,修复更为迅速。在正常组织中GAP-43表达量极少,骨骼肌损伤后各组中GAP-43表达量显著增高(P0.05);在第14天,GAP-43表达量达到高峰,D组继续表达呈上升趋势(P0.01),C组开始下降,但仍高于A组(P0.05)。与A组相比,骨骼肌受损后各时间点C、D两组agrin的表达明显增多(P0.05),其中D组差异最为显著(P0.01)。结论:电针有促进受损骨骼肌恢复的作用,可能与提高GAP-43和agrin的表达水平有关。     

生长相关蛋白在电针治疗大鼠脊髓损伤中的表达及意义

[目的]探讨生长相关蛋白(Gap-43)在电针治疗大鼠脊髓损伤(SCI)中的变化及意义.[方法]以2个月龄Wistar大鼠(96只)为受试对象,随机分为对照组(A组)、损伤组(B组)、治疗对照组(C组)和电流刺激治疗组(D组).B、C、D组于T9处行脊髓全横断,D组予电针治疗.用Western-blot测定各组损伤段脊...     

神经生长因子预处理阿尔茨海默病大鼠海马神经原磷酸化Tau蛋白的变化

背景：神经生长因子能够促进胆碱能神经元的分化,决定轴突的生长,并可参与损伤神经的再生和功能修复。目的：进一步验证神经生长因子预处理对拟阿尔茨海默病模型大鼠脑内神经原纤维缠结和磷酸化Tau蛋白表达的影响。方法：将3-5月龄雄性Wistar大鼠随机分为3组：模型组将冈田酸微量注射至拟大鼠海马CA1区建立拟阿尔茨海默病大鼠模型;预处理组于造模前将神经生长因子注射至侧脑室;对照组用同样方法注入等体积的二甲亚砜作为对照。通过Morris水迷宫观察上述大鼠的行为学变化,分别用改进的Bielschowsky染色观察海马CA1区神经原纤维缠结,用免疫组织化学和免疫印迹法观察海马区磷酸化Tau蛋白表达的变化。结果与结论：拟阿尔茨海默病模型组大鼠出现认知、学习记忆能力减退;与对照组比较,模型组海马CA1区出现较多神经原纤维缠结,而且磷酸化的Tau蛋白表达增多;神经生长因子预处理组大鼠的上述症状明显改善。提示神经生长因子预处理可以显著改善拟阿尔茨海默病模型大鼠的学习记忆能力,抑制神经原纤维缠结的形成,减少磷酸化Tau蛋白的表达。     

电针结合重复经颅磁刺激对局灶性脑缺血大鼠蛋白激酶A-环磷腺苷反应元件结合蛋白信号转导通路的影响

Objective To investigate the effects of electro-acupuncture (EA) combined with repetitive transcranial magnetic stimulation (rTMS) on protein kinase A-cyclic adenosine monophosphate response element binding protein (PKA-CREB) signal transduction system after focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Methods The animal model of transient focal ischemia was made by artificial middle cerebral artery occlusion. Seventy-five Wistar rats were randomly divided into normal group, model group, EA group, rTMS group and EA+rTMS group. The expressions of PKA and pCREB in hippocampus were detected and the neurologic impairment rating was observed at the 7th, 14th and 28th days, respectively, after infarction.Results The average gray densities of PKA and pCREB expressions in hippocampus after focal cerebral ischemia in model group were higher at the 7th d, lower at the 28th d than that in normal group (P＜0.05);higher in EA group, rTMS group, EA+rTMS group than that in model group at all time points (P＜0.05), higher in EA+rTMS group than that in EA group and rTMS group at 7th and 14th d(P＜0.05), and there was no difference between EA group and rTMS group(P＞0.05).The improvement of neural motor function was obvious in EA group, rTMS group and EA+rTMS group compared with model group (P＜0.01), especially in EA+rTMS group.Conclusions EA combined with rTMS can promote the functional recovery after cerebral ischemia,enhance the expression of PKA-CREB signal transduction system in hippocampus after focal cerebral ischemia, which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.     

电针结合重复经颅磁刺激对局灶性脑缺血大鼠蛋白激酶A-环磷腺苷反应元件结合蛋白信号转导通路的影响

目的探讨电针结合重复经颅磁刺激(rTMS)对局灶性脑缺血大鼠蛋白激酶A-环磷腺苷反应元件结合蛋白（PKA-CREB）信号转导通路的影响及其治疗缺血性脑损伤的机制。 方法取雄性Wistar大鼠75只,采用线栓法制备大鼠大脑中动脉闭塞模型,分为正常组、模型组、电针组、rTMS组和电针+rTMS组, 每组大鼠15只,通过蛋白印迹法检测正常组入组后及其它各组脑缺血后第7,14,28天3个时间点大鼠海马胞核内蛋白激酶A（PKA）、磷酸化环磷腺苷反应元件结合蛋白(pCREB)表达的变化,并观测其神经功能评分。 结果脑缺血后不同时间点缺血侧海马PKA及pCREB灰度值比较,模型组在造模后第7天时高于正常组,造模后第28天时低于正常组,差异均有统计学意义(P<0.05),造模后第14天时与正常组相比差异无统计学意义 (P&rt;0.05);电针组、rTMS组和电针+rTMS组3个时间点均高于模型组,造模后第7,14天时高于正常组,差异具有统计学意义(P<0.05),造模后第28天时与正常组相比差异无统计学意义(P&rt;0.05),其中,电针+rTMS组第7,14天时高于电针组、rTMS组,差异有统计学意义(P<0.05),电针组和rTMS组各时间点差异均无统计学意义(P&rt;0.05)。电针组、rTMS组和电针+rTMS组各时间点神经功能评分均较模型组改善(P<0.01),其中以电针+rTMS组神经功能评分改善最为明显。 结论电针结合rTMS对脑卒中后神经功能的恢复具有显著的促进作用, 蛋白激酶A-环磷腺苷反应元件结合蛋白信号转导通路蛋白的表达增强可能是其治疗缺血性脑卒中的机制之一。     

电针结合重复经颅磁刺激对局灶性脑缺血大鼠蛋白激酶A-环磷腺苷反应元件结合蛋白信号转导通路的影响

Objective To investigate the effects of electro-acupuncture (EA) combined with repetitive transcranial magnetic stimulation (rTMS) on protein kinase A-cyclic adenosine monophosphate response element binding protein (PKA-CREB) signal transduction system after focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Methods The animal model of transient focal ischemia was made by artificial middle cerebral artery occlusion. Seventy-five Wistar rats were randomly divided into normal group, model group, EA group, rTMS group and EA+rTMS group. The expressions of PKA and pCREB in hippocampus were detected and the neurologic impairment rating was observed at the 7th, 14th and 28th days, respectively, after infarction.Results The average gray densities of PKA and pCREB expressions in hippocampus after focal cerebral ischemia in model group were higher at the 7th d, lower at the 28th d than that in normal group (P＜0.05);higher in EA group, rTMS group, EA+rTMS group than that in model group at all time points (P＜0.05), higher in EA+rTMS group than that in EA group and rTMS group at 7th and 14th d(P＜0.05), and there was no difference between EA group and rTMS group(P＞0.05).The improvement of neural motor function was obvious in EA group, rTMS group and EA+rTMS group compared with model group (P＜0.01), especially in EA+rTMS group.Conclusions EA combined with rTMS can promote the functional recovery after cerebral ischemia,enhance the expression of PKA-CREB signal transduction system in hippocampus after focal cerebral ischemia, which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.     

G protein and adenylate cyclase complex-mediated signal transduction in the rat heart during sepsis

Changes in the protein level of various subunits of GTP-binding protein and the activity of adenylate cyclase in the rat heart during different phases of sepsis were studied. Sepsis was induced by cecal ligation and puncture (CLP). Experiments were divided into three groups: control, early sepsis, and late sepsis. Early and late sepsis refers to those animals sacrificed at 9 and 18 h, respectively, after CLP. The protein levels of various subunits of GTP-binding protein were determined by Western blot analysis. The activity of adenylate cyclase was measured based on the rate of formation of cAMP from [alpha-32P]ATP. The results show that protein levels of G alphas and G beta remained stable during the early and the late phases of sepsis. The protein levels of G alpha i-2 and G alpha i-3 remained relatively unaltered during the early phase of sepsis, but they were increased by 46.5% (P < 0.05) and 61.3% (P < 0.01), respectively, during the late phase of sepsis. The basal adenylate cyclase activity remained unchanged during the early phase while it was decreased by 25.7% (P < 0.05) during the late phase of sepsis. The isoproterenol-stimulated adenylate cyclase activity was unchanged during early sepsis while it was decreased by 44.6% (P < 0.01) during late sepsis. These data demonstrate that during the late hypodynamic phase of sepsis, myocardial G alpha i-2 and G alpha i-3 protein levels were increased and the increases were coupled with a reduction in adenylate cyclase activity. Because GTP-binding proteins mediate sympathetic control of cardiac function, the present findings may have a pathophysiological significance in contributing to the understanding of the pathogenesis of cardiac dysfunction during the late stage of sepsis.     

Prognostic evaluation of the protective potential of gastric mucosa in stomach ulcer

Potentialities of medical iconics in assessment of a protective mucosal barrier in gastric ulcer have been studied. Prognostic criteria have been derived for a common and a torpid course of the disease. Pathogenetic mechanisms of the above forms are specified.