人参茎叶皂苷对热损伤大鼠不同脏器糖皮质激素受体的影响

目的:观察人参茎叶皂苷(ginsenosides,GSS)对热损伤大鼠不同脏器糖皮质激素受体(glucocorticoidreceptor,GR)的影响,并探讨其作用机制。方法:雄性SD大鼠随机分为:(1)正常对照组,室温下饲养,蒸馏水灌胃;(2)GSS治疗组,室温下饲养,GSS灌胃;(3)热损伤模型组,蒸馏水灌胃,制作热损伤模型;(4)热损伤模型GSS治疗组,GSS灌胃,制作热损伤模型。采用放射配体结合法检测大鼠脑、胸腺、肺和肝细胞液GR结合活性;逆转录聚合酶链反应法测定脑、肝细胞液GRmRNA的水平;放射免疫法测定血浆促肾上腺皮质激素(adrenocorticotropin,ACTH)和皮质酮(corticosterone,CS)的浓度。结果:热损伤模型GSS治疗组大鼠脑、肺和肝细胞液GR结合活性以及脑和肝细胞液GRmRNA表达水平均明显高于单纯热损伤模型组(P<0.05或P<0.01);热损伤模型GSS治疗组大鼠血浆ACTH和CS浓度与单纯热损伤模型组比较则无明显差异。结论:GSS可缓解热损伤大鼠不同脏器GR结合活性的下降幅度,其作用机制可能与促进GRmRNA的表达有关。     

参附汤对休克大鼠不同器官糖皮质激素受体的上调作用

目的：探讨参附汤临床益气回阳救逆功效的作用机制。方法：以失血性休克大鼠为模型，观察参附汤对失血性大鼠血浆皮质酮(GC)、促肾上腺皮质激素(ACTH)及脑、肝、胸腺等部位糖皮质激素受体(GCR)的影响。结果：参附汤对失血性休克大鼠脑、肝、胸腺等部位的糖皮质激素受体活性有明显的上调作用。结论：参附汤通过保护GCR，提高机体GC系统在失血性休克过程中的生物学效应，可能是其临床益气回阳救逆功效的重要作用机制之一；参附汤上调失血性休克大鼠糖皮质激素受体的作用未见器官特务性。     

参附汤对失血性休克大鼠糖皮质激素及其受体的影响

深入探讨参附汤的药理作用机理。以急性失血性休克大鼠为实验对象，同步观察参附汤对血浆皮质酮及肝胞液，胸腺细胞糖皮质激素受体的影响。参附汤组大鼠肝胞液及胸腺细胞ＧＲ的结合位点都明显高于单纯失血组，参附汤组血浆皮质酮略高于失血组。参附汤可纠正失血性休克模型的ＧＲ的减少，以发挥其救治休克，回阳固脱的作用。     

生脉散组分配伍对热损伤大鼠糖皮质激素受体的影响

目的：研究生脉散组分配伍对热损伤大鼠糖皮质激素受体（glucocorticoid receptor,GR）的影响。 方法：雄性SD大鼠32只随机分为正常对照组、模型组、人参总皂苷组和生脉散组分配伍组（简称组分配伍组）,每组8只,连续灌胃1周进行预处理。除正常对照组外,其余各组大鼠于末次给药30min后复制大昂热损伤模型。随后立即断头处死动物,收集血清和肝、肺、肾脏组织。酶联免疫吸附测定法（enzyme-linked immkinosorbent assay,ELISA）检测血清皮质酮（corticosterone,CS）的浓度,放射免疫检测法（radioimmunoassay,RIA）检测血清促肾上腺皮质激素（adrenocorticotropic hormone,ACTH）的浓度;放射性配体受体结合法（radioligand receptor binding assay,RRA）检测肝、肺、肾细胞液GR结合容量。 结果：模型组肝、肺、肾胞液GR明显低于正常对照组（P〈0.01）。组分配伍组肝和肺胞液GR结合容量明显高于模型组（P〈0．01）,肾胞液GR结合容量与模型组比较,差异无统计学意义。组分配伍组肝胞液GR明显高于人参总皂苷组（P〈0．01）,肺和肾胞液GR结合容量与人参总皂苷组比较有上升的趋势．但差异无统计学意义。正常大鼠CS及ACTH水平与模型组、组分配伍组和人参总皂苷组比较,差异有统计学意义（P〈0．01）,模型组CS及ACTH水平与各给药组比较,差异无统计学意义。 结论：生脉散组分配伍可增强人参总皂苷上调热损伤大鼠GR水平的作用。     

参芪注射液对大鼠失血性休克再灌注肠粘膜糖皮质激素受体的调节作用

目的 探讨参芪注射液对大鼠失血性休克-再灌注后肠粘膜组织糖皮质激素受体的影响.方法 雄性SD大鼠72只,随机分成:正常组、模型组、参芪低剂量组(参芪注射液10g/kg)和参芪高剂量组(参纸注射液20g/kg).对SD大鼠静脉使用肝素后缓慢放血,建立重度血失性休克及复苏的动物模型;参芪注射液于再灌注前静脉注入.造模完成后观察各组动物肠粘膜病理学变化,动态观察2,6、12 h肠粘膜、GR的结合活性和GRRNA的表达水平.结果 正常组肠粘膜无明显改变,模型组肠粘膜重度损伤,参芪高剂量组轻度损伤,低剂量组中度损伤;参芪高剂量组与低剂量组比较,肠粘膜损伤减轻程度更明显.模型组与正常组比较,2、6、12 h肠粘膜GR结合活性和GRmRNA表达明显降低(P<0.01);参芪注射液高低剂量组与模型组比较,2、6、12h肠粘膜GR结合活性和GRmRNA表达明显升高(P<0.01,P<0.05);参芪高剂量组与低剂量组比较,2、6、12h肠粘膜GR结合活性和GRmRNA表达升高更明显(P<0.05).结论 参芪注射液能有效地减轻大鼠失血性休克再灌注肠粘膜损伤,提高肠粘膜GR结合活性和GRmRNA的表达水平.     

烫伤后大鼠肝糖皮质激素受体减少的机理——肾上腺部分切除动物的研究

部分去除肾上腺的大鼠烫伤后,血清糖皮质激素无明显升高,而肝胞液糖皮质激素受体(GR)却有升高的趋势,其中烫伤后6 h组和对照组的相差有非常显著的意义。进一步用打点杂交进行的研究表明,部分去腺大鼠烫伤后6h肝GR mRNA也有增加。这些结果提示,完整动物烫伤时肝胞液GR的减少依赖于血中糖皮质激素的升高。     

加味生脉散对热损伤大鼠糖皮质激素及其受体的影响

目的；探讨加味生脉散增强机体热耐力的作用机理。方法；以ＳＤ大鼠制备损伤模型，观察加味生脉散夺热损伤大鼠血浆糖皮质激素（ＧＣ）、促肾上腺皮质激素（ＡＣＴＨ）以及脑、肝胞洽谈以质激素受体（ＧＣＲ）的影响。结果；中味生脉散组大鼠脑、肝胞液ＧＣＲ的结合活性（Ｒｓ）（５３．１７±１４．０６，３３２．２３±５３．８３）炕于单纯员伤组（２８．２７±１０．２７，２１６．９０±５８．９７），同时并不降低血浆ＧＣ和Ａ     

休克肠淋巴液引流对大鼠红细胞代谢的作用

目的 观察休克肠淋巴液引流对失血性休克大鼠红细胞三磷酸腺苷(ATP)、乳酸(LA)、2,3-二磷酸甘油酸(2,3-DPG)以及红细胞内外离子浓度的影响,揭示肠淋巴途径在休克发病学中的意义.方法 Wistar雄性大鼠均分为假休克组、休克组(复制失血性休克模型)、引流组(复制失血性休克模型,自低血压1h引流休克肠淋巴液).在低血压3h或相应时间,经腹主动脉取血,制备红细胞悬液检测红细胞ATP、LA水平;制备红细胞内液检测2,3-DPG、Na+、K+浓度;制备血浆,检测Na+、K+、Cl-、Ca浓度.结果 休克组、引流组大鼠红细胞2,3-DPG、LA含量显著高于、ATP含量显著低于假休克组,且引流组2,3-DPG、ATP含量高于休克组;休克组血浆仅K+浓度高于假休克组、引流组红细胞内液Na+浓度显著低于休克组.结论 失血性休克大鼠红细胞能量代谢障碍的表现为2,3-DPG代偿性增加、ATP减少、LA堆积,并引起血钾升高;休克肠淋巴液引流增强了2,3-DPG的代偿、提高了ATP生成、减少LA堆积.肠淋巴液在休克红细胞能量代谢障碍的发病学中发挥重要作用.     

糖皮质激素受体在创伤失血性休克大鼠肝损伤中的作用

目的 从组织受体水平探讨糖皮质激素受体(GR)在创伤失血性休克后肝组织中的变化及其对肝损伤的作用机制.方法 采用双侧股骨骨折伴失血性休克模型,并对GR进行阻断后再致伤,动态观察伤后8h大鼠肝组织GR、肝脏病理、肝功能生化指标、伤后8h大鼠死亡率等变化.肝组织GR采用免疫印迹法测定其蛋白含量,并进行计算机图像分析.结果 肝组织GR的蛋白含量在创伤失血性休克后1h即开始下降,2h明显低于正常对照组(P＜0.01),6h降至最低,8h仍显著低于正常;光镜下伤后4h-8h肝窦内少许淤血,有散在炎性细胞浸润;血清丙氨酸氨基转移酶(ALT)、总胆红素(TB)伤后4h开始增高,白蛋白下降;大鼠死亡率10%.GR阻断后再致双侧股骨骨折并失血性休克,光镜下伤后2h肝窦内即可见较多炎性细胞浸润,血清ALT、TB在伤后2h即有明显升高(P＜0.01),白蛋白明显下降(P＜0.01);大鼠死亡率明显升高(50%).结论 GR不足可导致严重创伤并失血性休克后肝损伤的发生;且减少越多,肝损害越重,死亡率越高.提示GR在严重创伤休克后肝组织细胞损伤与抗损伤机制方面可能起着重要作用.     

养阴抗毒胶囊2号对阴虚大鼠糖皮质激素受体mRNA水平的影响

目的:利用氢化可的松法建立阴虚大鼠模型,观察养阴抗毒胶囊2号对阴虚大鼠模型肝组织糖皮质激素受体(GR)mRNA水平的影响,探讨其减轻激素副作用的机理。方法:SD大鼠40只,随机分为空白对照组、阴虚模型组、养阴抗毒胶囊2号组、六味地黄丸组和桂附地黄丸组。在中药预处理及造模给药9天后处死,放射免疫方法检测各组血浆皮质醇(CORT)含量,RT-PCR半定量测定肝组织GR mRNA水平,并进行图像分析和数据处理。结果:与模型组相比,养阴抗毒胶囊2号组和六味地黄丸组大鼠血浆CORT含量均显著降低(P<0.05),同时肝组织GR mRNA水平均明显升高(P<0.05),并且养阴抗毒胶囊2号组优于六味地黄丸组(P<0.05);而桂附地黄丸组大鼠的血浆CORT含量和肝组织GR mR-NA水平均与模型组没有差别(P>0.05)。结论:养阴抗毒胶囊2号能明显降低阴虚大鼠模型血浆CORT水平,提高肝组织GR mRNA水平,提示它可能改善HPA轴功能亢进状态,从而减轻激素副作用。     

Increased hsp70 of glucocorticoid receptor complex induced by scald and heat stress and its possible effect on the affinity of glucocorticoid receptor

Background Glucocorticoid （GC） insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor （GR）. Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins （hsp） especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro. Methods The affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting. Results Similar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats （30% total body surface area immersion scald） and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols. Conclusions The primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.     

Increased hsp70 of glucocorticoid receptor complex induced by scald and heat stress and its possible effect on the affinity of glucocorticoid receptor

Background Glucocorticoid (GC) insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor (GR). Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins (hsp) especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro. Methods The affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting.Results Similar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats (30% total body surface area immersion scald) and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols.Conclusions The primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.     

Increased hsp70 of glucocorticoid receptor complex induced by scald and heat stress and its possible effect on the affinity of glucocorticoid receptor

Background Glucocorticoid (GC) insensitivity/GC resistance is an important etiological and prognostic factor in multiple diseases and pathophysiological processes such as scald, shock and asthma. The function of GC was mediated by glucocorticoid receptor (GR). Scald not only decreased the expression of GR but also reduced the affinity of GR, which played an important role in GC resistance in scalded rats. Whereas the molecular mechanism responsible for the decrease of GR affinity resulted from scald remains unclear. Recent studies showed that the changes of heat shock proteins (hsp) especially hsp90 and hsp70 of GR heterocomplex were associated with GR low affinity in vitro. Methods The affinity of GR in hepatic cytosols and in the cytosols of SMMC-7721 cells were determined by radioligand binding assay and scatchard plot. GR heterocomplex in cytosols were captured by coimmunoprecipation and the levels of hsp90 and hsp70 of GR complex were detected by quantitative Western blotting.Results Similar with that of hepatic cytosol of scalded rats, a remarkable decrease of GR affinity was also found in the cytosol of heat stressed SMMC-7721 cells. The level of hsp70 of GR complex in hepatic cytosol of scalded rats (30% total body surface area immersion scald) and in cytosol of heat stressed human hepatocarcinoma cell line SMMC-7721 were both increased by 1.5 fold, whereas no change of hsp90 in GR heterocomplex was found. According to the correlation analysis, there may be a positive relationship between increased hsp70 of GR complex and decreased GR affinity in the cytosols.Conclusions The primary results indicated that the level of hsp70 of GR heterocomplex was increased in the hepatic cytosol of scalded rats and the cytosol of heat stressed SMMC-7721 cells. The increase of hsp70 of GR complex might be associated with the decrease of GR affinity.     

甘草酸二铵上调急性肺损伤大鼠糖皮质激素受体的表达

目的观察甘草酸二铵(DG)对急性肺损伤(ALI)大鼠肺组织糖皮质激素受体(GR)及血清肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)表达的影响。方法将36只雄性SD大鼠随机分为假手术组(S组)、ALI组、DG干预组(HLG组)、地塞米松干预组(HLD组)、ALI受体阻断组(ALIR组)和DG干预受体阻断组(HLGR组)。各组动物造成失血性休克,然后给予内毒素(LPS)2mg/kg腹腔注射;LPS注射前1h,分别给予DG 20mg/kg或地塞米松2mg/kg腹腔注射,ALIR组和HLGR组大鼠肌肉注射受体阻断剂RU486。测定大鼠肺湿干比(W/D)、血氧分压(PaO2),并检测肺组织GR mRNA及GR蛋白的表达,同时测定血清TNF-α、IL-10浓度。结果 (1)ALI组TNF-α明显高于S组、HLGR组和HLG组(P<0.01);ALI组IL-10明显高于S组(P<0.01),但低于HLG组和HLGR组(P<0.05)。(2)ALI组GR mRNA和蛋白的表达明显低于S组(P<0.01);HLG组GRmRNA和蛋白的表达则明显高于ALI组(P<0.05,P<0.01)。(3)ALI组肺组织出现充血、水肿、中性粒细胞浸润,HLG组肺部炎症较ALI组减轻。结论 DG可能通过上调GR及IL-10的表达,同时抑制TNF-α的过度表达,从而起到减轻ALI的作用。     

The glucocorticoid resistance in burn and shock

The changes of glucocorticoid receptor (GR) and the reactivity of target cells toglucocorticoids (GC) were studied experimentally.The results showed that GC resistance,which wascaused mainly by the decrease of GR,developed in burned and shocked rats and dogs.Thesechanges may exacerbate the shock state,and even lead to the development of multiple organfailuue.The protection of GC against intestinal ischemic shock of dogs was demonstrated after theconcentration of dexamethasone (Dex) in plasma was elevated to 10~(-6) mol/L by iv injection of 5mgDex/kg body weight.     

孕期激素对后代大鼠脑发育的影响

目的通过对妊娠期孕鼠产前应用不同疗程糖皮质激素(GCs),研究后代仔鼠脑组织重量、含水量及糖皮质激素受体的表达,了解孕期使用糖皮质激素对后代脑发育的影响。方法孕鼠于怀孕第18天开始每天肌注地塞米松0.48 mg/(kg·次),q4 h一次,4次为一疗程,连用3 d;单疗程后期与对照组以同等容积生理盐水代替。仔鼠生后第1天(P1)、P3称脑重,用干湿法测脑含水量;于P3、P7、P42用ISH法测量糖皮质激素受体mRNA在脑海马部位的表达。结果仔鼠胎儿期应用激素后脑重减轻,脑组织含水量却无改变,糖皮质激素受体表达下调。结论胎儿时期使用过量糖皮质激素影响仔鼠脑发育。