热毒清对内毒素所致家兔DIC氧自由基代谢的影响

采用内毒素所致家兔弥散性血管内凝血(DIC)为模型,观察热毒清是否具有抗自由基作用。实验表明,与模型组比较,热毒清组血清、肝匀浆和线粒体LPO含量以及血清和肝匀浆XOD活性均显著降低;而肝匀浆SOD以及肝匀浆和线粒体GSH-Px活性明显升高。结果提示热毒清能有效地拮抗内毒素DIC 病理过程中增多的自由基,并通过抑制自由基生成酶和增强自由基清除酶活性而起作用。从而部分解释了热毒清保护细胞器的作用机理。     

中医清法制剂“热毒清”抗内毒素的实验研究

热毒清 (抗炎6号) 体外能使鲎试验转阴,电镜下可见能使大肠杆菌O_(111)B_4内毒素网状结构崩解。体内实验用家兔内毒素所致DlC模型,检测血小板、纤维蛋白原、凝血酶原时间和3F试验;光镜检查肾小球微血栓形成率和肾小球所含纤维素密度等,用药组和模型组比较差异均非常显著(P<0.01)。结合电镜观察和肝细胞溶酶体标志酶ACF测定,提示热毒清抗内毒素的作用机理可能与①直接降解内毒素,②增强MPS吞噬功能,③稳定溶酶体膜有关。 ·     

热毒清的拆方研究──对DIC家兔肝线粒体呼吸功能和体外稳定溶酶体膜的影响

对组成复方热毒清的４味中药进行拆方研究，分别观察其对播散性血管内凝血（ＤＩＣ）模型家兔肝细胞线粒体呼吸功能和体外稳定溶酶体膜的影响。结果表明：线粒体呼吸控制率（ＲＣＲ），模型组明显低于热毒清组、地塞米松组和正常组（Ｐ＜０．０１）；但组成热毒清的４味中药单方各项指标与模型组比较无明显统计学意义（Ｐ＞０．０５）。在体外稳定溶酶体膜实验中，除热毒清组溶酶体标志酶酸性磷酸酶（ＡＣＰ）和β－葡萄糖苷酸酶在溶酶体上清液中之酶比活性较低（Ｐ＜０．０５）外，余４味中草药均无统计学意义。由此可见：复方热毒清在保护ＤＩＣ家兔肝线粒体损害和体外稳定溶酶体膜方面优于组方中各单味药，从而从一个侧面验证了热毒清组方的合理性。     

心脉灵液对内毒素休克大鼠肝损伤保护作用研究──光镜电镜观察及线粒体定量分析

本实验观察了心脉灵液对内毒素休克大白鼠肝脏的组织学及超微结构变化，并对肝细胞线粒体进行了定量分析。光镜下可见内毒素休克组大白鼠肝血窦及中央静脉扩张瘀血，伴有白细胞增多、聚集，肝细胞肿胀、胞浆嗜酸性变，空泡变性；透射电镜可见肝细胞严重损伤，包括核固缩，线粒体肿胀、嵴消失、部分基质浓缩，内质网扩张，糖原含量减少。线粒体图象分析表明，内毒素休克组肝细胞线粒体数密度，面密度及表面积分体积之比明显降低，平均表面积及平均体积增大，与正常对照组相比Ｐ＜０．０１，心脉灵治疗组以上指标明显改善，与内毒素休克组相比Ｐ＜０．０１。     

热毒清的拆方研究：对DIC家兔肝线粒体呼吸功能和体外稳定…

对组成复方热毒清的４味中药进行拆方研究，分别观察其对播散性血管内凝血模型家兔肝细胞线粒体呼吸功能和体外稳定溶酶体膜的影响。     

热毒清对红细胞膜Na~+,K~+-ATP酶保护作用的实验研究

热毒清(原名抗炎6号)系我校附属同济医院用四味清热解毒中药制成的静脉注射液,具有抑菌、抗炎、解毒、退热、增强免疫功能及稳定溶酶体膜、保护线粒体H~+-ATP合酶等多方面的作用,其多相药理作用可能与保护生物膜活性有关。本实验研究热毒清对内毒素所致家兔红细胞膜Na~+,K~+-ATP酶活性损害作用的影响,以探讨热毒清治疗疾病的机理。 一、材料和方法 1.动物、健康日本大耳白兔24只,体重2～2.5 kg, 雌雄不拘,随机分为4组,每组6只。①正     

中医清热解毒法的实质研究获新成果

同济医科大学附属同济医院对中医清热解毒法制剂“热毒清”抗内毒素和保护溶酶体、线粒体实验研究通过鉴定,省内外有关中医、西医、中西医结合专家们认为,此项科研成果达到国际先进水平。 “热毒清”(原名抗炎6号)是用四味清热解毒中药制成的静脉注射液,经8个医院的共同临床观察,证明对多种急性感染性疾病疗效满意,总有效率在85％以上,某些急腹症疗效也达82.5％。为了进一     

内毒素及肿瘤坏死因子对肝细胞超微结构的影响

目的：研究内毒素及肿瘤坏死因子对肝细胞超微结构的影响。方法：我们采用胶原酶二步法灌注一月龄ＳＤ雄性大鼠肝脏,制备肝细胞悬液并进行了原代培养,重点研究了内毒素、肿瘤坏死因子对原代短期培养肝细胞超微结构的影响。结果：表现为糖原减少、溶酶体增多、线粒体肿大、嵴溶解、内质网扩张及断裂、甚至可见核空泡化、染色质消失。其中以肿瘤坏死因子引起的肝细胞损伤为重;若二者联合作用,则肝细胞受损更加严重。结论：内毒素与     

黄芪对梗阻性黄疸幼鼠肝脏超微结构及IL-10 mRNA表达的影响

目的 建立幼鼠梗阻性黄疸模型,对模型鼠应用黄芪干预,观察胆道梗阻后肝脏超微结构和IL-10 mRNA表达的改变,探讨黄芪对上述指标的影响.方法 将40只Wistar幼鼠随机分为正常对照组、假手术组、梗阻性黄疸(OJ)组和OJ+黄芪(A)组,每组10只.OJ组鼠于近肝门处游离并结扎胆总管,假手术组仅做游离不结扎胆总管.OJ+A组术后1d开始腹腔注射黄芪注射液(250 mg/100 g体重)共7 d,其余组相同时间腹腔注射生理盐水0.5 ml.检测实验鼠肝脏超微结构改变;应用RT-PCR技术检测肝组织IL-10 mRNA表达,对所得数据进行统计学处理.结果 (1)电镜下正常对照和假手术组肝细胞结构完整,细胞器分布均,线粒体正常.Disse间隙内可见枯否细胞,OJ组肝细胞内线粒体明显肿胀、减少、外膜不清、嵴消失,粗面内质网数量减少,滑面内质网增多、核糖体脱颗粒.肝细胞内质网扩张,细胞核边聚,毛细胆管扩张.OJ+A组肝细胞的超微结构变化较轻.(2)IL-10 mRNA基因表达OJ组显著高于对照组及OJ+A组,均P＜0.05.结论 梗阻性黄疸幼鼠模型肝脏超微结构显著改变;幼鼠胆管结扎8 d,肝组织的IL-10 mRNA表达增强;应用黄芪的梗阻性黄疸幼鼠肝细胞超微结构改变较轻,减轻肝脏IL-10 mRNA的表达,从而对梗阻性黄疸诱发的肝脏损害有保护作用.以上结果证实黄芪可较好的改善梗阻性黄疸机体的营养状况,对受损的肝脏起保护作用.     

非酒精性脂肪性肝炎大鼠肝脏内源性H_2S合成减少

目的探讨非酒精性脂肪性肝炎(nonalcoholic steatohepatitis,NASH)大鼠肝脏硫化氢(hydrogen sulfide,H2S)变化及与肝细胞线粒体损伤的可能关系。方法采用雄性SD大鼠26只,分为正常对照组(n=6),NASH模型组(n=10)及复方牛胎肝提取物治疗组(n=10),高脂饮食12周建立NASH大鼠模型。亚甲基蓝分光光度法测定肝组织H2S生成率,RT-PCR及免疫组织化学测定肝组织CSEmRNA及其表达。透射电子显微镜观察肝细胞超微结构、PCR法测定肝细胞线粒体DNA(mtDNA)片段ND1、ND6、CO1、CYB及ATP6。结果 NASH大鼠模型组肝细胞线粒体肿大、膜模糊或嵴消失,正常组大鼠mtDNA片段ND1、ND6、CO1、CYB及ATP6表达明显高于模型组和治疗组(P<0.05)。NASH模型组肝组织H2S生成率〔(1.26±0.08)nmol/min〕较正常组〔(2.11±0.17)nmol/min〕明显减少(P<0.05),且肝脏CSEmRNA水平及其表达减少,治疗组与模型组比较差异无统计学意义。结论 NASH大鼠肝脏H2S合成减少可能与肝细胞线粒体损伤有关,H2S在NASH发病机制中可能具有保护肝脏的作用。     

Studies on the stabilizing effect of traditional Chinese preparation - Re Du Qing - on the membrane of lysosomes

Summary This experiment which include in vitro and in vivo experiment suggested that Re Du Qing could protect the lysosomes of the rabbit liver cells. It may be further suggested that Re Du Qing can prevent and alleviate endotoxic DIC by stabilizing the membrane of lysosomes.     

Experimental studies on the antagonistic effect of re du qing on endotoxin-induced damage of lysosomes and mitochondria

A rabbit endotoxic DIC model was preliminarily performed. The structures of lysosomes and mitochondria in liver cells were evidently destroyed as observed under electronic microscope, whereas those rabbits pretreated with Re Du Qing (RDQ, formerly named anti-inflammatory No. 6)--a mixture of Chinese traditional herbs providing antipyretic and detoxifying action, showed principally normal ultrastructure in liver cells. In lysosomal functional studies, the activity of the lysosomal marker enzyme--acid phosphate (ACP) was 79.0 +/- 4.7% (M +/- SD) in model group, higher than pretreated group (54.01 +/- 4.0%, P less than 0.01). Studies on the mitochondrial function showed that the significant criteria of the respiratory activity of mitochondria--respiratory control ratios (RCR) was 2.83 +/- 1.08 in model group and markedly lower than pretreated group (5.46 +/- 1.25, P less than 0.01). Mitochondrial ATPase activity (mu mol pi/min/mg pr.) was lower in model group (0.280 +/- 0.015) than in pretreated group (0.341 +/- 0.018, P less than 0.05). Lipid peroxide (LPO) in liver homogenates and serum were 1.86 +/- 0.43 n mol MDA/mg pr. and 12.26 +/- 0.84 n mol MDA/ml respectively in model group, whereas in pretreated group they gave a much lower value (1.19 +/- 0.12 and 6.55 +/- 2.97) respectively. Those data showed very significant difference between two groups (P less than 0.01). All of the above indices of pretreated group yielded values close to those of normal control group. The results of experimental study in vitro were identical to those of experimental study in vivo. These experimental studies suggested that RDQ provide antagonistic effect on endotoxin induced damage of lysosomes and mitochondria.     

Preventive effect of re du qing on hepatocytes and mitochondria damaged by lipid peroxidation in experimental rabbits with endotoxin-induced disseminated intravascular coagulation

In this study, the general Shwartzman reaction of rabbits induced by Escherichia Coli endotoxin was made as DIC models. The experiments showed that the levels of lipid peroxide (LPO) in hepatic tissue and mitochondria in the model group were increased significantly compared with the control group (P less than 0.01), while superoxide dismutase (SOD) activity in hepatic tissue and glutathione peroxidase (GSH-Px) activity in hepatic tissue and mitochondria were decreased significantly (P less than 0.01). The levels of LPO in hepatic tissue and mitochondria in Re Du Qing (RDQ) group and vitamin E (VE) group were decreased significantly (P less than 0.01 and P less than 0.05 respectively) compared with the model group. The levels of LPO in the RDQ group did not differ from the control group (P greater than 0.05), but the levels of LPO in the VE group were still higher than those in the control group significantly (P less than 0.05). The SOD activity in hepatic tissue and GSH-Px activity in hepatic tissue and mitochondria in both RDQ group and VE group were also significantly higher than those in the model group (P less than 0.01). These data suggest that the levels of oxygen free radicals were increased in hepatocytes and mitochondria. This is related to the decreased activities of SOD and GSH-Px in the course of pathogenesis of endotoxin-induced DIC. This study indicates that lipid peroxidation might be one of the important mechanisms resulting in hepatocellular and mitochondria from oxidative damage.     

肝毒清颗粒治疗慢性重型肝炎合并内毒素血症的临床疗效评价

目的：观察肝毒清颗粒治疗慢性重型肝炎合并内毒素血症的临床疗效。方法：将49例慢性重型肝炎合并内毒素血症患者随机分成两组。其中25例以肝毒清颗粒配合西医综合治疗作为治疗组,24例以单纯西医综合治疗作为对照组。观察治疗后的总有效率,以及内毒素、肝功能和凝血酶原活动度的变化。结果：治疗组总有效率（76%）优于对照组（45.8%）,差异有统计学意义（P﹤0.05）。两组均可改善肝功能,升高凝血酶原活动度,清除血浆内毒素,治疗组与对照组比较,差异有统计学意义（P＜0.01）。结论：肝毒清颗粒治疗慢性重型肝炎合并内毒素血症临床疗效显著。     

热毒清与抗生素联用治疗小儿肺炎的临床疗效和T细胞亚群的观察

中药热毒清结合抗生素(简称中西药组)和单用抗生素(称抗生素组)分别治疗小儿肺炎各30例。结果表明:中西药组的主要症状、体征如气喘、咳嗽,紫绀、肺部哮鸣音和中小湿罗音等消失,均较抗生素组为早(P<0.05);前组痊愈率为100.0％,明显高于后组的83.33％(P<0.05)。治疗前,两组患儿的OKT~+_4细胞皆较正常值为低,OKT~+_8细胞皆较正常为高(P<0.01)。经治疗后,两组OKT~+_4细胞都可恢复正常,而OKT~+_8细胞仅在中西药组恢复正常。     

Experimental study of the protective action of re-du-qing on H(+)-APT synthase activity of mitochondria in the rabbit

To observe the effect of Re-Du-Qing on the toxic effect of endotoxin, the rabbit model of general Shwartzman reaction was made by injection of endotoxin into ear vein and the H(+)-ATP synthase activities of rabbit liver mitochondria were measured. The results showed that the H(+)-ATP synthase activity of endotoxin group was significantly lower than that of the normal group and group which had been simultaneously injected Re-Du-Qing (P less than 0.05). The level of H(+)-ATP synthase activity was almost raised to normal value, when Re-Du-Qing or dexamethasone was injected simultaneously with endotoxin respectively. It suggested that Re-Du-Qing was able to alleviate the nocuous effect of endotoxin on H(+)-ATP synthase activity in the mitochondria of rabbit liver cells.     

清热利湿解毒汤对类风湿性关节炎活动期的临床及实验研究

以清热利湿解毒汤为主治疗58例主要病证为湿热毒的类风湿性关节炎活动期病人,17例西药对照。中药组有效率为96.6％,西药组为58.8％。实验证明清热利湿解毒汤具有抑制体液免疫,清除血清免疫复合物,抗炎、降温作用。     

肝性脑病动物模型的鉴定指标分析

目的:通过对慢性肝功能衰竭引起肝性脑病(hepat-ic encephalopathy,HE)发生模型的多指标观察和分析,寻找理想的HE模型的特征指标。方法:应用四氯化碳联合内毒素法制作慢性肝功能衰竭引起HE发生的模型,进行行为活动观察,血液生化、形态学、迷宫试验等检测。结果:模型组大鼠在造模过程中先后有67%(10/15)的动物死亡,出现呆滞、步态蹒跚、疼痛反应迟钝甚至消失等神经系统功能异常表现,并伴有血氨升高趋势,总胆红素、直接胆红素、γ-谷氨酰转肽酶升高(P<0.05),血浆蛋白、钙、钠和氯显著降低(P<0.01)。电镜检查显示模型组大鼠肝脏有纤维化性变化,海马脑片的神经元有趋向死亡性变化。完成造模的模型组大鼠与对照组相比,迷宫实验的测试结果没有显著性差异。结论:用四氯化碳联合内毒素法制作的慢性肝功能衰竭引起HE发生的模型,行为变化明确,但其他检测指标具有多样性和复杂性,需做进一步的综合性分析。