云南灯盏花胶囊对慢性肾功能衰竭大鼠TGF-β_1与PAI-1 mRNA表达及肾功能的影响

目的:研究中药复方制剂云南灯盏花胶囊对慢性肾功能衰竭(CRF)大鼠肾功能及肾组织纤维化的影响。方法:60只SD大鼠,随机分为正常对照组、模型组、苯那普利组和灯盏花组。应用Platt 5/6肾切除方法制备CRF模型。苯那普利组予盐酸苯那普利(0.29 mg/100 g)灌胃,灯盏花组予灯盏花胶囊(0.3 g/100 g)灌胃,正常组及模型组予等量生理盐水灌胃,疗程12周。检测各组大鼠血肌酐、尿素氮、24 h尿蛋白定量;RT-PCR方法检测各组大鼠肾组织肿瘤生长因子-β1(TGF-β1)、血浆纤溶酶原激活物抑制剂-1(PAI-1)的mRNA表达。结果:灯盏花组及苯那普利组大鼠血清肌酐、尿素氮浓度和24 h尿蛋白定量与模型组比较均明显降低(P<0.05),且灯盏花组大鼠血清肌酐、24 h尿蛋白定量均低于苯那普利组(P<0.05)。灯盏花组、苯那普利组大鼠肾组织TGF-β1和PAI-1 mRNA表达均低于模型组,差异有统计学意义(P<0.05),且灯盏花组TGF-β1和PAI-1 mRNA表达水平低于苯那普利组(P<0.05)。结论:云南灯盏花胶囊能改善CRF大鼠的肾功能,抑制肾组织纤维化。

病毒性心肌炎患儿外周血中TNF-α、TGF-β1和IFN-γ细胞因子水平分析

目的:探讨病毒性心肌炎患儿外周血中肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)和细胞因子干扰素-γ(IFN-γ)与疾病的关系。方法:以临床确诊的28例病毒性心肌炎患儿为研究对象,与20例正常健康儿童进行比较,采用Elisa方法检测血清中TNF-α、TGF-β1和IFN-γ蛋白含量;采用实时荧光定量PCR法检测TNF-α、TGF-β1和IFN-γmRNA水平。结果:与健康儿童相比较,病毒性心肌炎患儿外周血IFN-γ的蛋白含量及IFN-γmRNA表达水平明显降低(P<0.01);TNF-α、TGF-β1的蛋白含量及TNF-α、TGF-β1的mRNA表达水平明显升高(P<0.01)。结论:TNF-α、TGF-β1和IFN-γ参与病毒性心肌炎的发病过程。

E-钙黏蛋白在神经母细胞瘤上皮-间质转化中的作用

目的研究神经母细胞瘤中E-钙黏蛋白(E-cadherin)的表达在肿瘤转化生长因子-β1(TGF-β1)诱导的上皮-间质转化(EMT)中的作用。方法体外采用TGF-β1(1μg/L、5μg/L、10μg/L)处理人神经母细胞瘤SK-N-SH细胞株,与空白对照组比较,采用荧光定量PCR及Western blot法检测EMT相关基因及蛋白表达。采用迁移实验及划痕实验检测肿瘤细胞迁移能力的改变。选取2008年1月至2012年12月上海交通大学附属儿童医院血液肿瘤科收治的神经母细胞瘤患儿18例。收集患儿术后肿瘤组织,采用免疫组织化学法检测E-cadherin的表达情况,并与患儿临床特点及生存预后行相关性分析。结果与空白对照组相比,SK-N-SH细胞经TGF-β1(1μg/L、5μg/L、10μg/L)处理后,实时荧光定量PCR及Western blot均提示上皮细胞标志E-cadherin的mRNA(0.603±0.081、0.606±0.008、0.716±0.166比1.000)及蛋白(0.855±0.026、0.600±0.017、0.495±0.011比1.000)表达量明显下降(F=8.144,P=0.035;F=74.810,P<0.001),间质细胞标志α-平滑肌肌动蛋白的mRNA(2.132±0.167、3.494±0.017、4.184±0.021比1.000)及蛋白(1.175±0.053、1.189±0.058、1.225±0.106比1.000)的表达量明显上升(F=547.300,P<0.001;F=68.810,P=0.007),提示发生EMT。划痕实验及Transwell迁移实验均发现随着TGF-β1处理后(5μg/L),迁移细胞数明显增多(t=16.070,P=0.040)。神经母细胞瘤患儿组织病理中E-cadherin强阳性、阳性、弱阳性及阴性表达的10年总体生存(OS)率分别为(77.78±13.86)%、(75.00±21.66)%、(25.00±21.65)%及0,差异有统计学意义(F=8.160,P=0.040)。结论TGF-β1可诱导神经母细胞瘤SK-N-SH细胞发生EMT,并使细胞迁移能力增强。神经母细胞瘤患儿E-cadherin表达降低与疾病临床进展及复发/转移明显相关。

Effect of electroacupuncture at conception vessel and governor vessel on correlative signals of stress-injury-repair in cerebral ischemia reperfusion rats

Objective To explore the effect of electroacupuncture at conception vessel and governor vessel on correlative signals of inflammatory response stressinjury-repair in cerebral ischemia reperfusion rats. Methods A total of 216 rats were included in the experiment, of which 192 rats were randomly divided into four groups, including sham operation group（group A）, model group（group B）, governor vessel group（group C）, and conception vessel and governor vessel group（group D）, 48 rats in each group. The rats in each group were further divided into six sub-groups, including 12 h group, 24 h group, 48 h group, 72 h group, 96 h group and 144 h group. MCAO operation course was simulated in group A, and the common carotid artery, internal carotid artery and external carotid artery on the right side were exposed, without insertion of suture or treatment. After completion of modeling in group B, treatment was not conducted. After completion of modeling in group C, ＂Shuǐgōu（水沟 GV 26）＂, ＂Bǎihuì（百会 GV 20）＂ and ＂Dàzhuī（大椎 GV 14）＂ of governor vessel were selected, and electroacupuncture device was connected with ＂GV 14＂ and ＂GV 20＂. After completion of modeling in group D, ＂GV 26＂, ＂GV 20＂ and ＂GV 14＂ of governor vessel, and ＂Qìhǎi（气海 CV 6）＂, ＂Guānyuán（关元 CV 4）＂ and ＂Chéngjiāng（承浆 CV 24）＂ of conception vessel were selected; electroacupuncture device was connected with ＂GV 14＂ and ＂GV 20＂, ＂CV 4＂ and ＂CV 6＂. Neurological severity score was conducted.Enzyme linked immunosorbent assay（ELISA） was applied to determine the concentration of peripheral serum adrenocorticotropic hormone（ACTH） of rats, and immunohistochemical method was adopted to detect the expression levels of tumor necrosis factor α（TNF-α） and transforming growth factor β1（TGF-β1） in brain tissue of rats. The other 24 rats were also randomly divided into four groups mentioned above, 6 rats in each group, to detect cerebral ischemie volume. Results Compared with group B, the ischemia volume in brain tissue at 144 h after cerebral ischemia reperfusion decreased significantly in group C and group D（P〈0.05）, and the ischemia volume in group D was less than that in group C（P〈0.05）. The neurological impairment scores of group C and group D at all time points were lower than those of group B, and the differences were statistically significant（all P〈0.05）. The scores of group D at all time points were lower than those of group C, and the differences since 24 h were statistically significant（all P〈0.05）. Compared with group B, the excessive expression of ACTH was suppressed dynamically and generally in group C and group D（P〈0.05）; the expression level of TNF-α was down-regulated and TGF-β1 was up-regulated as well in group C and group D（P〈0.05）. The effect in group D was superior to that in group C. Conclusion Electroacupuncture at conception vessel and governor vessel can protect brain through the intervention of the expression of correlative signals of inflammatory response stress-injury-repair of cerebral ischemia reperfusion.