墨旱莲对4种蝮蛇毒引起的炎症和出血的影响

目的探讨墨旱莲提取液对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒所致的炎症和出血的影响。方法应用短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒所致大鼠足跖肿胀的致炎模型,观察墨旱莲提取液对蛇毒所致大鼠足跖肿胀的影响。墨旱莲提取液分别与不同蛇毒混合,给小鼠腹部皮下注射,观察其对蛇毒引起的小鼠皮下出血的影响。结果墨旱莲提取液15g／kg连续2次灌胃给药,对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒或尖吻蝮蛇毒所致大鼠足跖肿胀的急性炎症造模和短尾蝮蛇毒棉球肉芽肿的慢性炎症造模(20g／kg)均有明显的抑制作用,对这些蛇毒引起的小鼠皮下出血也能明显抑制。结论墨旱莲提取液对短尾蝮蛇毒、蛇岛蝮蛇毒、白眉蝮蛇毒及尖吻蝮蛇毒引起的炎症和出血均有明显的抑制作用。     

中国的毒蛇和抗蛇毒血清

我国产有近五十种毒蛇中,具有剧毒且对人畜危害较大者,约有十几种,即银环蛇,眼镜蛇,金环蛇,眼镜王蛇,蝰蛇,蝮蛇,尖吻蝮,竹叶青,烙铁头蛇及常见的几种海蛇。我国抗蛇毒血清的研制,一直未得到应有的重视,及至1970年始有抗蛇毒血清供临床使用。目前计有四种产品,即精制抗蝮蛇毒血清,精制抗尖吻蝮蛇毒血清,精制抗银环蛇毒血清和精制抗眼镜蛇毒血清。现正陆续研制抗蝰蛇毒血清,抗金环蛇毒血清等,为治疗我国毒蛇咬伤中毒患者提供可靠的特效药。     

尖吻蝮蛇蛇毒出血毒素的纯化与部分性质

目的　被尖吻蝮蛇 （ Ｄｉｅｎａｇｋｉｓｔｒｏｄｏｎ ａｃｕｔｕｓ） 咬伤会引起严重的出血, 对蛇毒出血毒素的研究有利于治疗蛇伤出血药物筛选。 方法　采用 Ｓｅｐｈａｄｅｘ Ｇ７５, Ｄ Ｅ Ａ Ｅ Ｓｅｐｈａｄｅｘ Ａ５０, Ｓｅｐｈａｄｅｘ Ｇ２００ 和两次 Ｐ Ｂ Ｅ 聚焦层析纯化。 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 电泳和等电聚焦电泳测定纯化样品的纯度和等电点。氨基酸组成用自动氨基酸分析仪测定。以小鼠背部皮下注射部位出血斑的面积来确定最小出血剂量和常规的方法测定酶活性。结果　从尖吻蝮蛇毒中纯化到一个相对分子量为５６ ０００ 的出血毒素 （ Ｄａ Ｈ Ｔ３）, 经氨基酸组成测定计算,它由 ４８７ 个氨基酸残基组成。此成分在 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ上显示出一条均一的蛋白染色带, 其ｐ Ｉ为５５０。该出血成分的最小出血剂量是 ２６μｇ, 具有蛋白水解酶活力, 其活力为 ３６８, 但没有精氨酯酶和磷脂酶 Ａ２ 活力。当加入 Ｅ Ｄ Ｔ Ａ 螯合剂去除金属离子后, 它们的出血活力和蛋白水解酶活力均丧失。 结论　这是从大陆尖吻蝮蛇毒中获得的一个新的出血金属蛋白酶 （ Ｄａ Ｈ Ｔ３）。     

尖吻蝮蛇毒对小鼠肾脏损伤的研究

尖吻蝮(五步蛇)Agkistrodonacutus是我国特有的毒蛇,尖吻蝮蛇毒除引起局部出血肿胀和坏死等症状外,临床常伴有肾功能衰竭或尿毒症的症状。为给临床治疗提供依据,本实验对尖吻蝮蛇毒引起损伤后肾脏的改变,用小鼠作了试验观察,材料与方法取体重20克左右的小白鼠。每只注入40微克的尖吻蝮蛇毒,先将粗蛇毒溶子生理盐水中,使每0.1毫升生理盐水含     

蛇油化学成分的研究

蛇油的应用在我国有悠久的历史,李时珍在《本草纲目》中记载了蛇油的功效:蛇脂“绵裹塞耳聋,亦傅肿痛”,又“摩着物皆透也”。民间常用蛇油治疗冻疮、皲裂、湿疹及水火烫伤。蛇油的现代研究,已见文献及产品。作者以尖吻蝮蛇油为原料制成了“黄山蛇油膏”,用于临床治疗冻疮显效率753%,总有效率94%,治疗手足皲裂显效率90%,总有效率98%。为深入研究蛇油成分,更好地开发蛇油制品,作者对尖吻蝮蛇油及以混合蛇油为原料制成的“黄山蛇油膏”的主要脂肪酸进行了定性、定量分析。1　实验材料尖吻蝮蛇油　取成体尖吻蝮10条,放血致死,取蝮腔内脂肪,洗净…     

五步蛇毒毒中蛋白凝集素的发现及其凝集特性的研究

发现尖吻蝮蛇(Agkistrodon acutus)粗毒对动物细胞有凝集作用。顺序用DEAE—Sephadlex A-50,CM Sephadex C25柱层析,从尖吻蝮蛇毒中分离纯化出一种毒蛋白凝集素。经分子量,等电点和出血活性等实验测定确定该凝集素就是徐询等人早期分离、纯化的出血毒素Ⅲ(AaHⅢ)。这种凝集素对许多动物细胞都有凝集作用,例如脾淋巴细胞、HeLa细胞、人肾成纤维细胞和草鱼(Ctenoph argngodon idellus)。但对组细胞和血小板无作用。和植物凝集素PHA相比AaHⅢ产生细胞的凝集块体积要大些。对淋巴细胞起凝集作用的最低浓度为2.5ug/ml。能使HeLa细胞和淋巴细胞凝集在一起。如预先加肝素和溶菌酶可以抑制对淋巴细胞的凝集怍用。用溶菌酶(2mg/ml)和AaHⅢ(1mg/ml)在37℃′下保温2小时AaHⅢ的出血活性仍然存在。     

用尖吻蝮蛇毒蝮蛇毒及抗蛇毒血清处理小鼠S180,EAC腹水癌的初步观察

本文报道了尖吻蝮蛇毒、蝮蛇毒及抗蛇毒血清能使接种S180、EAC腹水癌的小鼠明显延长存活时间、降低接种率,但不能完全阻止癌细胞生长。体外具有较明显的导致癌细胞肿胀、膜破裂、核纤维化、坏死等。从腹水酶活力测定及抗血清初步研究结果表明,癌细胞病变中产生的某些抗原物质可能与蛇毒中的酶和毒蛋白相近。因此注射蛇毒后可在体内产生相关抗体,中和癌细胞产生的毒素以达到治疗目的。     

乌梢蛇血清的抗出血因子:一个有前途的抗蛇毒药物原料

用柱层析和聚丙烯酰胺凝胶盘状电泳法,从乌梢蛇血清中分离纯化了一个抗出血因子。用SDS-聚丙烯酰胺凝胶电泳法测得其分子量大约为65 kD;测定了五种蝮亚科蛇毒(尖吻蝮、竹叶青蛇、原矛头蝮、哈扑和短尾蝮)的最小出血剂量和乌梢蛇血清中抗出血因子对这五种蛇毒的抗出血活性;还测定了七种蛇毒(除上述五种毒蛇外,还包括圆斑蝰和银环蛇)的半数致死量,以及抗出血因子对中毒小鼠的治疗作用。结果显示:从乌梢蛇血清中提纯的抗出血因子的抗蛇毒活性,不仅可以抵抗它的捕食者尖吻蝮的蛇毒,而且还可以抵抗具出血活性的其它蛇毒;但它对不具出血活性的银环蛇毒的致死抑制作用不明显。该抗出血因子不仅在体外实验表现出强的中和出血毒素的活性,而且在体内实验中亦表现出对中毒小鼠良好的治疗作用,因而可能成为新的抗蛇毒药物的有前途的原料。乌梢蛇血清对血循毒的中和能力的获得,可能归因于尖吻蝮与乌梢蛇之间捕食与被捕食相互作用的关系。     

中草药徐长卿(Cynanchum paniculatum)治疗毒蛇咬伤的分子机制研究

徐长卿(Cynanchum paniculatum)作为一种传统中草药,被广泛应用于毒蛇咬伤治疗,然而其疗效的科学实验证据缺乏,治疗的分子机制也不清楚.本文采用有机溶剂萃取、柱层析及高效液相色谱法(HPLC)相结合,从徐长卿中筛选出和分离得到一个功效组分,经质谱和核磁共振鉴定为丹皮酚原苷.动物活体实验与组织病理切片观察表明,该组分显著地抑制尖吻蝮(Deinagkistrodon acutus)蛇毒引起的出血、水肿及肌肉组织坏死等活性.体外酶活检测显示,丹皮酚原苷可以抑制尖吻蝮蛇毒水解纤维蛋白原活性和PLA2活性.进一步采用分子模拟软件技术,揭示出丹皮酚原苷是通过与蛇毒中主要成分金属蛋白酶和PLA2发生相互作用,从而抑制尖吻蝮蛇毒发挥致伤作用.本研究首次揭示了中草药徐长卿治疗蛇伤的分子机制,为中医治疗毒蛇咬伤使用徐长卿找到了科学实验证据,也为蛇伤药物的研发提供了新的思路.     

用尖吻蝮蛇毒、蝗蛇毒及抗蛇毒血清处理小鼠S_(180)、EAC腹水癌的初步观察

本文报道了尖吻蝮蛇毒、蝮蛇毒及抗蛇毒血清能使接种S_(180)、EAC腹水癌的小鼠明显延长存活时间、降低接种率,但不能完全阻止痛细胞生长。体外具有较明显的导致,菡细胞肿胀、膜破裂,核纤维化,坏死等。从腹水酶活力测定及抗血清初步研究结果表明,癌细胞病变中产生的某些抗原物质可能与蛇毒中的酶和毒蛋白相近。因此注射蛇毒后可在体内产生相关抗体,中和癌细胞产生的毒素以达到治疗目的。     

Potentiated Interaction between Ineffective Doses of Budesonide and Formoterol to Control the Inhaled Cadmium-Induced Up-Regulation of Metalloproteinases and Acute Pulmonary Inflammation in Rats

The anti-inflammatory properties of glucocorticoids are well known but their protective effects exerted with a low potency against heavy metals-induced pulmonary inflammation remain unclear. In this study, a model of acute pulmonary inflammation induced by a single inhalation of cadmium in male Sprague-Dawley rats was used to investigate whether formoterol can improve the anti-inflammatory effects of budesonide. The cadmium-related inflammatory responses, including matrix metalloproteinase-9 (MMP-9) activity, were evaluated. Compared to the values obtained in rats exposed to cadmium, pretreatment of inhaled budesonide (0.5 mg/15 ml) elicited a significant decrease in total cell and neutrophil counts in bronchoalveolar lavage fluid (BALF) associated with a significant reduction of MMP-9 activity which was highly correlated with the number of inflammatory cells in BALF. Additionally, cadmium-induced lung injuries characterized by inflammatory cell infiltration within alveoli and the interstitium were attenuated by the pre-treatment of budesonide. Though the low concentration of budesonide (0.25 mg/15 ml) exerted a very limited inhibitory effects in the present rat model, its combination with an inefficient concentration of formoterol (0.5 mg/30 ml) showed an enhanced inhibitory effect on neutrophil and total cell counts as well as on the histological lung injuries associated with a potentiation of inhibition on the MMP-9 activity. In conclusion, high concentration of budesonide alone could partially protect the lungs against cadmium exposure induced-acute neutrophilic pulmonary inflammation via the inhibition of MMP-9 activity. The combination with formoterol could enhance the protective effects of both drugs, suggesting a new therapeutic strategy for the treatment of heavy metals-induced lung diseases.     

Tumor necrosis factor-induced lethal hepatitis: pharmacological intervention with verapamil, tannic acid, picotamide and K76COOH

Tumor necrosis factor (TNF) induces hepatitis when injected in human beings or in rodents. The molecular mechanism by which TNF induces hepatic distress remains largely unknown, although induction of apoptosis of hepatocytes appears to be an essential step. In order to increase the therapeutic value of TNF, we have studied the protective activity of several molecules and found that four chemically totally different substances confer significant protection in the model of TNF-induced lethal hepatitis in mice sensitized with D-(+)-galactosamine (GalN), but not in mice sensitized with actinomycin-D (ActD) or against anti-Fas-induced lethal hepatitis. Verapamil, a calcium-channel blocker, tannic acid, picotamide, a thromboxane A(2) receptor antagonist, and K76COOH, an inhibitor, amongst others, of complement, protected significantly against induction of lethality, release of the liver-specific enzyme alanine aminotransferase (ALT) and induction of apoptosis in the liver after TNF/GalN, except for K76COOH, which paradoxically increased ALT values after challenge, and which also protected against TNF/GalN in complement-deficient mice. The data suggest that activation of platelets and neutrophils, as well as induction of inflammation occur in the TNF/GalN model, but not in the TNF/ActD or anti-Fas models, in which direct induction of apoptosis of hepatocytes may be more relevant. The protective activity of the drugs may lead to an increase in therapeutic value of TNF.     

Protective Effects of Selenium on Cadmium-Induced Brain Damage in Chickens

Selenium (Se) is an important dietary micronutrient with antioxidative roles. Cadmium (Cd), a ubiquitous environmental pollutant, is known to cause brain lesion in rats and humans. However, little is reported about the deleterious effects of subchronic Cd exposure on the brain of poultry and the protective roles on the brain by Se against Cd. The aim of this study was to investigate the protective effects of Se on Cd-induced brain damage in chickens. One hundred twenty 100-day-old chickens were randomly assigned to four groups and were fed a basal diet, or Se (as 10 mg Na₂SeO₃/kg dry weight of feed), Cd (as 150 mg CdCl₂/kg dry weight of feed), or Cd?+?Se in their basic diets for 60 days. Then, concentrations of Cd and Se, production of nitric oxide (NO), messenger RNA (mRNA) level and activity of inducible NO synthase (iNOS), level of oxidative stress, and histological and ultrastructural changes of the cerebrum and cerebellum were examined. The results showed that Cd exposure significantly increased Cd accumulation, NO production, iNOS activities, iNOS mRNA level, and MDA content in the cerebrum and cerebellum. Cd treatment obviously decreased Se content and antioxidase activities and caused histopathological changes in the cerebrum and cerebellum. Se supplementation during dietary Cd obviously reduced Cd accumulation, NO production, mRNA level and activity of iNOS, oxidative stress, and histopathological damage in the cerebrum and cerebellum of chickens. It indicated that Se ameliorates Cd-induced brain damage in chickens by regulating iNOS-NO system changes, and oxidative stress induced by Cd and Se can serve as a potential therapeutic for Cd-induced brain lesion of chickens.     

Cellular glutathione peroxidase protects mice against lethal oxidative stress induced by various doses of diquat

This study was to determine if cellular glutathione peroxidase (GPX1) protects against acute oxidative stress induced by diquat. Lethality and hepatic biochemical indicators in GPX1 knockout mice [GPX1(-/-)] were compared with those of wild-type mice (WT) after an intraperitoneal injection of diquat at 6, 12, 24, or 48 mg/kg of body weight. Although the WT survived all the doses, the GPX1(-/-) survived only 6 mg diquat/kg and were killed by 12, 24, and 48 mg diquat/kg at 52, 4.4 and 3.9 hr, respectively. Compared with those of surviving mice that were sacrificed on Day 7, the dead GPX1(-/-) had diquat dose-dependent increases (P < 0.05) in plasma alanine aminotransferase (ALT) activities. The GPX1(-/-) also had higher (P < 0.05) liver carbonyl contents than those of the WT, but the differences were irrespective of diquat doses. Whereas hepatic total GPX and phospholipid hydroperoxide glutathione peroxidase activities or hepatic GPX1 protein was not significantly affected by the diquat treatment, liver thioredoxin reductase and catalase activities were lower (P < 0.05) in the GPX1(-/-) injected with 12 mg diquat/kg than those of other groups. In conclusion, normal GPX1 expression is necessary to protect mice against the lethality, hepatic protein oxidation, and elevation of plasma ALT activity induced by 12-48 mg diquat/kg.     

Flavonoids protect mice from two types of lethal shock induced by endotoxin

The protective effect of flavonoids on two types of lethal endotoxic shock was studied. A lethal endotoxic shock was induced by administration of lipopolysaccharide (LPS) into D-galactosamine (D-GalN)-sensitized mice and another one was done by administration of a high dose of LPS into normal mice. Pretreatment with a series of flavonoids protected mice from two types of endotoxin lethality. Flavonoid pretreatment reduced the serum tumor necrosis factor-alpha (TNF-alpha) level in mice injected with D-GalN and LPS, but not in mice injected with a high dose of LPS. TNF-alpha-induced lethal shock in D-GalN-sensitized mice was also protected by pretreatment with flavonoids, suggesting that flavonoids augmented the resistance to TNF-alpha lethality. On the other hand, flavonoids reduced the plasma level of lipid peroxides in mice injected with a high dose of LPS, but not in D-GalN-sensitized mice. Taken together, these results indicated that flavonoids might protect mice from two types of endotoxin lethality. The protective mechanism of flavonoids in each endotoxin lethality is discussed.     

Differential response of a(2)-macroglobulin-deficient mice in models of lethal TNF-induced inflammation

Tumor necrosis factor (TNF) is an essential mediator in the pathogenesis of Gram-negative septic shock. Injection of TNF into normal mice leads to systemic, lethal inflammation, which is indistinguishable from lipopolysaccharide (LPS)-induced lethal inflammation. alpha(2)-macroglobulin (A2M) is a major positive acute phase protein with broad-spectrum protease-inhibitory activity. Mouse A2M-deficient (MAM-/-) mice were significantly protected against lethal systemic inflammation induced by TNF. The protection is not due to faster clearance of the injected TNF. The induction of tolerance to TNF-induced lethality by repetitive administration of small doses of human TNF for five consecutive days was equally efficient in both mutant mice compared to wild-type mice. In D-(+)-galactosamine (GalN)-sensitized mice, TNF induces lethal inflammatory hepatitis. MAM(-/-) mice are equally sensitive to the lethal combination of TNF/GalN. Furthermore, interleukin-1-induced desensitization to TNF/GalN was not impaired in MAM(-/-) mice. We conclude that MAM plays a mediating role in TNF-induced lethal shock and that MAM deficiency does not reduce changes in efficiency of tolerance and desensitization to TNF and TNF/GalN-induced lethality, respectively.     

Requirement of endogenous tumor necrosis factor/cachectin for recovery from experimental peritonitis

By intrasplenic immunization we raised a rat mAb (mAb V1q; IgG2a, kappa) with a potent neutralizing activity against natural mouse TNF (1 microgram/ml mAb V1q/100 U/ml TNF). mAb V1q was used to study the role of endogenous TNF in experimental peritonitis induced by sublethal cecal ligation and puncture. mAb V1q persisted for over 5 days in the serum of mice injected with 100 micrograms of the antibody and, therefore, proved useful for in vivo experiments. As little as 20 micrograms mAb V1q/mouse prevented lethal shock of the animals by 400 micrograms LPS/mouse. In sublethal cecal ligation and puncture i.p. injection of mAb V1q directly and up to 8 h after induction of experimental peritonitis lead to death of the animals within 1 to 3 days. The lethal effect of mAb V1q was compensated by injection of recombinant mouse TNF. Similar mAb V1q effects as in immunocompetent mice were shown in severe combined immune deficiency mice deficient of mature functional B and T cells. Taken together, these data suggest that during the early phase of peritonitis endogenous TNF may stimulate nonlymphoid cells such as granulocytes, macrophages, platelets, and fibroblasts to ingest bacteria and to localize inflammation, respectively. These beneficial effects of TNF may determine survival. Thus, our data may have implications for the therapeutic management of a beginning peritonitis.     

新型硫化氢供体GYY4137对脂多糖诱导的小鼠急性肺损伤的保护作用

目的:急性肺损伤是临床上常见的危重病,发病急,死亡率高,目前仍缺乏有效的治疗手段,新型的外源性硫化氢供体GYY4137具有抗炎、抗休克、抗癌及抗血栓等作用,本研究探讨其对脂多糖诱导的小鼠急性肺损伤的保护作用及其机制。方法:将BALB/c小鼠(18-20 g)随机分为3组:正常对照组(20只),脂多糖组(20只),治疗组(20只),然后复制小鼠脂多糖诱导的急性肺损伤模型:给予小鼠腹腔注射脂多糖(10 mg/kg)复制小鼠急性肺损伤模型模型,治疗组注射脂多糖1小时后给予腹腔注射GYY4137(50 mg/kg),在给予脂多糖8小时后将小鼠处死,留取血清与组织标本。检测小鼠血清中的炎症因子肿瘤坏死因子α、白介素6及白介素10的表达,检测小鼠血清中H2S的含量,测得肺脏湿/干比,检测肺组织中的髓过氧化物酶活性,并测得肺组织中与氧化应激相关的H2O2、·OH与SOD因子的含量。结果:脂多糖引起了严重的肺损伤,GYY4137对脂多糖导致的肺水肿、炎症反应及氧化应激损伤有不同程度的改善,保护了脂多糖造成的肺损伤,降低了脂多糖诱导的小鼠肺脏氧化应激损伤。其保护作用于抗炎、抗氧化有关。结论:GYY4137可能通过抗炎、抗氧化作用途径保护了脂多糖造成的急性肺损伤,可能在炎症疾病模型中也发挥相同作用,并且为未来临床使用缓释硫化氢供体提供了基础资料。     

Protective effects of sodium butyrate against lung injury in mice with endotoxemia

The aim of the present study was to investigate the effects of sodium butyrate (SB) on systemic inflammation, lung injury and survival rate of mice with endotoxemia. Balb/c mice were pre-treated with SB or vehicle, and then endotoxemia was induced by lethal dose of lipopolysaccharide (LPS, 20 mg/kg, i.p.) and the survival rate of mice was monitored. A separated set of animals were sacrificed at 18 h after LPS challenge, and blood samples were harvested for measuring TNF-α and IL-6 levels. Lung tissues were also harvested to determine the ratio of wet weight to dry weight of lung tissue and myeloperoxidase (MPO) activity in lung tissue. In addition, the formalin-fixed lung specimens were stained with HE routinely for morphologic evaluation. The results showed that pre-treatment with SB alleviated LPS-induced morphological damage in lung tissue. This was accompanied by reduced ratio of wet weight to dry weight of lung tissue and MPO activity in lung homogenates. Additionally, the up-regulation of pro-inflammatory cytokines TNF-α and IL-6 was also suppressed by SB, while the survival rate of mice with lethal endotoxemia was significantly increased by SB pre-treatment. The results suggest that SB effectively attenuates intrapulmonary inflammatory response and improves the survival of endotoxemic mice.     

Protective and therapeutic efficacy of Lactobacillus casei against experimental murine infections due to Mycobacterium fortuitum complex

A bacterial immunopotentiator, LC 9018 (heat-killed Lactobacillus casei), was studied for its protective and therapeutic efficacies against Mycobacterium fortuitum and M. chelonae infections in mice. This agent reduced the incidence of spinning disease and gross renal lesions and enhanced the elimination of organisms at the site of infection in the host mice, when administered intramuscularly six times a week (0.1 mg dry weight per injection, one injection on each day of treatment) from 1 week before to 2 weeks after infection. The LC 9018 injections in this protocol caused a marked increase in the phagocytic function, O2- -producing ability and chemiluminescence of host peritoneal macrophages. Moreover, LC 9018 injections using the same schedule resulted in an enhancement of interleukin-1-producing function of the macrophages, particularly in the infected mice. These findings indicate that LC 9018 administration with the present protocol can activate macrophage functions, in particular those related to microbicidal activity. This would partly explain the protective and therapeutic efficacy of LC 9018 against infection due to M. fortuitum complex.