目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理...目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理组(SNI+RS102895)和小胶质细胞活化抑制剂米诺环素(MC)处理组(SNI+MC),共4组。其中假手术组和模型组均进一步分为7 d、14 d和18 d组,SNI+RS102895和SNI+MC组均在第18天时取材。手术诱发神经痛,采用机械缩足阈(PWTs)测定不同时点痛阈,所有小鼠处死前2 d进行高架十字迷宫(EPM)实验,处死前1 d进行旷场实验(OFT)观察焦虑样行为变化。免疫荧光法检测小鼠海马CA3脑区白细胞分化抗原45(CD45)的表达及与小胶质细胞标记物离子钙接头蛋白分子1(IBA-1)、跨膜蛋白119(TMEM119),星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)和神经元标记物神经元核抗原(NeuN)的共表达,流式细胞术检测14 d SNI小鼠全脑单核细胞的百分比。18 d小鼠SNI后第5~16天分别灌胃给予MC 90 mg/(kg·d)、RS1028955 mg/(kg·d)与生理盐水,观察阻断单核细胞浸润对小鼠神经痛、焦虑样行为及海马CA3区CD45及IBA-1的表达的影响。结果7 d和14 d组小鼠SNI后1 d PWTs下降,持续到处死前(P<0.01)。假手术组CD45表达极少;与同时段假手术组比较,7 d SNI小鼠CD45表达无增加((P>0.05),14 d SNI小鼠CD45表达显著上升(P<0.01),且只与IBA-1和TMEM119有少量共表达,与GFAP、NeuN无共表达。14 d SNI小鼠全脑单核细胞百分比显著上升(P<0.01)。抑制小胶质细胞激活与抑制CCR2表达均能减少SNI小鼠CA3区CD45的表达(P<0.01),且能提高SNI小鼠机械痛阈(P<0.01)并缓解焦虑样行为(P<0.01)。结论SNI诱发神经痛14 d后小鼠海马CA3区有外周单核细胞的浸润,该现象可能参与了神经痛的维持及促进焦虑样行为的发生。展开更多
Background: Systemic hypertension is one of the main risk factors for atheroth rombosis. Tissue factor(TF) is found in the adventitia of blood vessels and in t he lipid core of atherosclerotic plaques, and is specific...Background: Systemic hypertension is one of the main risk factors for atheroth rombosis. Tissue factor(TF) is found in the adventitia of blood vessels and in t he lipid core of atherosclerotic plaques, and is specifically expressed on monoc yte or macrophage cell membrane surfaces. TF plays a pivotal role in blood clott ing physiology and is involved in pro-inflammatory action and atherosclerotic p laque destabilization. Objective: In this study we investigated whether there is any relationship between TF messenger RNA expression and activity in blood mono cytes isolated from hypertensive patients with clinical signs of atherosclerosis ,uncomplicated hypertensive individuals and normotensive control subjects. Metho ds: Eighty subjects(41 men and 39 women, mean age 41±12 years)with untreated es sential hypertension and 41 control subjects matched for sex and age were enroll ed in the study. Patients were classified according to whether they had a normal (≤1 mm, 41 patients) or abnormal(>1 mm, 39 patients) intima-media thickness(IM T). Results: TF mRNA expression and activity in hypertensive individuals with no carotid atherosclerosis were no different from control subjects in unstimulated and stimulated monocytes. Abnormal IMT patients showed a higher TF mRNA express ion compared with normal IMT hypertensive subjects(P< 0.001). Conclusions: We de monstrated that TF mRNA and activity levels in monocytes obtained from uncomplic ated hypertensive individuals are comparable with those of normotensive subjects , whereas atherosclerotic hypertensive patients showed increased levels of these parameters.展开更多
文摘目的观察不同时间点神经痛小鼠海马CA3区外周单核细胞的浸润,阐述这一现象对小鼠神经痛及焦虑样行为的影响。方法采用健康雄性C57小鼠,将小鼠随机分为假手术组(sham)、坐骨神经分支选择性损伤(SNI)模型组(SNI)、CCR2抑制剂RS102895处理组(SNI+RS102895)和小胶质细胞活化抑制剂米诺环素(MC)处理组(SNI+MC),共4组。其中假手术组和模型组均进一步分为7 d、14 d和18 d组,SNI+RS102895和SNI+MC组均在第18天时取材。手术诱发神经痛,采用机械缩足阈(PWTs)测定不同时点痛阈,所有小鼠处死前2 d进行高架十字迷宫(EPM)实验,处死前1 d进行旷场实验(OFT)观察焦虑样行为变化。免疫荧光法检测小鼠海马CA3脑区白细胞分化抗原45(CD45)的表达及与小胶质细胞标记物离子钙接头蛋白分子1(IBA-1)、跨膜蛋白119(TMEM119),星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)和神经元标记物神经元核抗原(NeuN)的共表达,流式细胞术检测14 d SNI小鼠全脑单核细胞的百分比。18 d小鼠SNI后第5~16天分别灌胃给予MC 90 mg/(kg·d)、RS1028955 mg/(kg·d)与生理盐水,观察阻断单核细胞浸润对小鼠神经痛、焦虑样行为及海马CA3区CD45及IBA-1的表达的影响。结果7 d和14 d组小鼠SNI后1 d PWTs下降,持续到处死前(P<0.01)。假手术组CD45表达极少;与同时段假手术组比较,7 d SNI小鼠CD45表达无增加((P>0.05),14 d SNI小鼠CD45表达显著上升(P<0.01),且只与IBA-1和TMEM119有少量共表达,与GFAP、NeuN无共表达。14 d SNI小鼠全脑单核细胞百分比显著上升(P<0.01)。抑制小胶质细胞激活与抑制CCR2表达均能减少SNI小鼠CA3区CD45的表达(P<0.01),且能提高SNI小鼠机械痛阈(P<0.01)并缓解焦虑样行为(P<0.01)。结论SNI诱发神经痛14 d后小鼠海马CA3区有外周单核细胞的浸润,该现象可能参与了神经痛的维持及促进焦虑样行为的发生。
文摘Background: Systemic hypertension is one of the main risk factors for atheroth rombosis. Tissue factor(TF) is found in the adventitia of blood vessels and in t he lipid core of atherosclerotic plaques, and is specifically expressed on monoc yte or macrophage cell membrane surfaces. TF plays a pivotal role in blood clott ing physiology and is involved in pro-inflammatory action and atherosclerotic p laque destabilization. Objective: In this study we investigated whether there is any relationship between TF messenger RNA expression and activity in blood mono cytes isolated from hypertensive patients with clinical signs of atherosclerosis ,uncomplicated hypertensive individuals and normotensive control subjects. Metho ds: Eighty subjects(41 men and 39 women, mean age 41±12 years)with untreated es sential hypertension and 41 control subjects matched for sex and age were enroll ed in the study. Patients were classified according to whether they had a normal (≤1 mm, 41 patients) or abnormal(>1 mm, 39 patients) intima-media thickness(IM T). Results: TF mRNA expression and activity in hypertensive individuals with no carotid atherosclerosis were no different from control subjects in unstimulated and stimulated monocytes. Abnormal IMT patients showed a higher TF mRNA express ion compared with normal IMT hypertensive subjects(P< 0.001). Conclusions: We de monstrated that TF mRNA and activity levels in monocytes obtained from uncomplic ated hypertensive individuals are comparable with those of normotensive subjects , whereas atherosclerotic hypertensive patients showed increased levels of these parameters.
