双功能域重组FN多肽抑制癌细胞浸润能力的研究

在制备了两个Cell Ⅰ-Hep Ⅱ 双结构域重组FN多肽(CH50和CH56)的基础上,研究其抑制肿瘤细胞浸润能力的作用。两个多肽的结构差异是CH50中删除了Cell I和HepⅡ之间的Ⅲ-11和ED-A结构顺序。CH50(ED_(50)为30.2 nmol/L)结合细胞的能力略高于CH56(ED_(50)为45.4 nmol/L)。两种多肽均可显著抑制黑色素瘤B16/F1细胞结合层粘素的能力,抑制作用相同。在体内肿瘤浸润抑制试验中,两种多肽均可显著抑制癌细胞浸润能力,使肺转移结节数降低80％左右。结果提示:Ⅲ-11和ED-A结构顺序对Cell Ⅰ-Hep Ⅱ 双结构域多肽结合细胞的能力有一定的影响,但删除Ⅲ-11和ED-A不是重组多肽抑制肿瘤转移的决定因素,Cell I和Hep Ⅱ 这两个结构域单独连接在一起是其抑制肿瘤细胞转移的结构基础。     

Similarities between the relaxations induced by vasoactive intestinal peptide and by stimulation of the non-adrenergic non-cholinergic neurons in the rat stomach

Summary The characteristics of the non-adrenergic, noncholinergic inhibitory response of the rat stomach fundus to transmural nerve stimulation were compared with the relaxation induced by vasoactive intestinal polypeptide (VIP). Treatment with -chymotrypsin (5 U/ml) or VIP antiserum (1:200) significantly reduced the relaxation induced by transmural nerve stimulation at 30 Hz, indicating that the possible transmitter in the non-adrenergic, non-cholinergic nerves is a peptide and may be VIP or a closely related peptide. VIP was able to relax, fully and dose-dependently, the stomach fundus that had previously been constricted by treatment with 10^–6 M serotonin, and the IC₅₀ value for VIP was 2.4 × 10^–9 M. VIP elevated levels of cyclic AMP in a dose-dependent manner and the EC₅₀ value was 2.8 × 10^–9 M in the presence of 10^–6 M atropine and 10^–6 M guanethidine. The stomach fundus was relaxed by transmural nerve stimulation (30 Hz, 50 mA) and transmural nerve stimulation also caused production of cyclic AMP in the rat stomach in the presence of atropine and guanethidine. The basal level of cyclic AMP in the stomach was 8.7 ± 0.26 pmole/mg protein. When transmural nerve stimulation was applied for 5 min, the contraction of the stomach, induced by 10^–6 M serotonin, was inhibited by 54% in the presence of atropine and guanethidine and the level of cyclic AMP was increased to 13.0 ± 0.73 pmol/mg protein. Apamin inhibited the transmural nerve stimulation-induced relaxation and shifted the dose-response curve for VIP to the right. These results suggest that one of the putative neurotransmitter from non-adrenergic, non-cholinergic nerves in the rat stomach is VIP and that VIP-induced relaxation may be mediated by the production of cyclic AMP and by the opening of apamin-sensitive K⁺-channels.Send offprint requests to K. Kamata at the above address     

Human isolated small intestine: motor responses of the longitudinal muscle to field stimulation and exogenous neuropeptides

Summary (1) Longitudinal muscle strips from the human small intestine (jejunum/ileum) responded to electrical field stimulation (1–50 Hz) with frequency-related primary contractions which were largely atropine- (3 M) sensitive. When the tone was raised by addition of galanin (0.3 – 1 M), prostaglandin (PG) E₂ (1–10 M) or neurokinin A (NKA, 0.1 M), a frequency-related relaxation was evident which was potentiated by atropine. All the responses to field stimulation were abolished by tetrodotoxin (1 M), thus indicating their neural origin. (2) The atropine-sensitive primary contraction to field stimulation was virtually abolished by omega conotoxin fraction GVIA (CTX, 0.1–0.3 M) while the relaxations were CTX-resistant. The field stimulation-induced relaxations, which were observed in the presence of atropine and guanethidine (3 M), were also unaffected by apamin (0.1 M). (3) NKA and substance P (SP) produced a concentration- (1 nM–1 M for both peptides) related contraction, NKA being about 53 times more potent than SP. [Pro⁹]SP sulphone and [MePhe⁷]-NKB, selective agonists of the NK-1 and NK-3 receptor, respectively, were barely effective. On the other hand, [\Ala⁸]NKA(4–10), a selective NK-2 receptor agonist, had a potent contractile activity, similar to that of NKA. (4) Galanin (1 nM–1M) produced an atropine- and tetrodotoxin-resistant concentration-related contraction of longitudinal muscle of human isolated small intestine. The response to galanin did not show any sign of fading and was particularly suitable to study the evoked relaxations. (5) Calcitonin gene-related peptide (CGRP) (10–100 nM) consistently inhibited the nerve-mediated contractions of strips from the ileum while the effect on the jejunum was less pronounced. Vasoactive intestinal polypeptide (VIP, 0.1–1 M) inhibited nerve-mediated contractions both in the ileum and the jejunum. (6) These experiments indicate that both cholinergic excitatory and non-adrenergic non-cholinergic nerves affect motility of the longitudinal muscle of the human small intestine. Furthermore, several neuropeptides produce potent motor effects, the contractile response to tachykinins being apparently mediated by activation of NK-2 receptors.     

Effect of vasoactive intestinal polypeptide on the release of serotonin from the in vitro vascularly perfused small intestine of guinea pig

Summary Isolated segments of the guinea pig small intestine were vascularly perfused and the release of endogenous serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) into the portal vein was measured. All test substances were intraarterially perfused. Vasoactive intestinal polypeptide (VIP, 1 pmol/l — 100 nmol/1) inhibited the spontaneous release of 5-HT and 5-HIAA. The maximal inhibitory effect (about 60%) was seen at 100 pmol/1. The effect of VIP on the spontaneous release of 5-HT and 5-HIAA was not changed in the presence of 1 ol/l tetrodotoxin (TTX).Raising intraluminal pressure by 500 Pa for 5 min increased the release of 5-HT and 5-HIAA by about 25%. Raising the intraluminal pressure in the presence of VIP reduced the release of 5-HT and 5-HIAA by about 75%. In the presence of TTX (1 gmol/l), raising intraluminal pressure also caused a decrease of the release of 5-HT and 5-HIAA which was unaffected by the additional presence of VIP. The fluid volume expelled during peristaltic activity was not affected by VIP, but reduced by about 90% in the presence of TTX.In conclusion the results demonstrate a direct inhibitory effect of VIP on the release of 5-HT from the enterochromaffin cells. In addition, VIP appears to interfere with the neuronally mediated stimulation of 5-HT release during peristaltic activity. Send offprint request to H. Schwörer at the above address     

日本猴椎旁交感神经节中含肽神经元的研究

本实验系应用荧光免疫组织化学的方法观察猴下位腰段椎旁交感神经节(L_(6-7))中神经肽Y,血管活性肠肽,降钙素基因相关肽,和P物质的存在、分布情况以及它们与酪氨酸羟化酶的共存关系。结果表明,大量细胞呈神经肽Y免疫反应阳性,它们在神经节周边分布更为密集。中等数量的血管活性肠肽阳性细胞和小量降钙素基因相关肽细胞散在于神经节内。在经含有Colchiciue的培养液离体孵育12h的标本上,可见中等数量的P物质免疫反应阳性细胞。根据抗酪氨酸羟化酶(TH)抗体的免疫染色结果,神经节内的神经元可分为TH~+和TH~-两群,前者占大多数。相邻切片免疫染色结果表明,几乎所有神经肽Y免疫阳性细胞同时含有TH,而所有血管活性肠肽免疫反应阳性细胞均呈酪氨酸羟化酶免疫反应阴性。神经肽Y与血管活性肠肽无共存关系。降钙素基因相关肽存在于部分血管活性肠肽免疫反应阳性细胞中,即属于VIP~+/TH~-组。从以上结果得出结论,在猴下位腰段椎旁交感神经节中,神经肽Y与血管活性肠肽分别存在于TH~+和TH~-两个细胞群。即神经肽Y存在于TH阳性神经元中,血管活性肠肽和降钙素基因相关肽则存在于TH阴性神经元中。     

蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其机制

目的 探讨蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其抗肿瘤机制.方法 四甲基偶氮唑蓝(MTT)比色法检测蝎毒重组蛋白对人肺癌A549细胞增殖的抑制作用;荧光显微镜及流式细胞仪(FCM)技术观测蝎毒重组蛋白对肿瘤细胞内钙离子浓度([Ca2+]I)的影响,探讨[Ca2+]I变化时Ca2+的可能来源.结果 蝎毒重组蛋白对人肺癌A549细胞生长有明显抑制作用;荧光显微镜显示,蝎毒重组蛋白作用细胞48 h后细胞内钙离子荧光强度增强,提示[Ca2+]I升高;流式细胞仪显示蝎毒重组蛋白在不同情况下可显著增加肿瘤[Ca2+]I(P<0.05).结论 蝎毒重组蛋白对人肺腺癌A549细胞增殖具有显著抑制作用,可能与升高肿瘤[Ca2+]I继而诱导肿瘤细胞凋亡有关;其升高肿瘤[Ca2+]I是通过开放肿瘤细胞膜钙通道和肿瘤细胞内钙库释放两条途径实现.     

蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其机制

目的 探讨蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其抗肿瘤机制.方法 四甲基偶氮唑蓝(MTT)比色法检测蝎毒重组蛋白对人肺癌A549细胞增殖的抑制作用;荧光显微镜及流式细胞仪(FCM)技术观测蝎毒重组蛋白对肿瘤细胞内钙离子浓度([Ca2+]I)的影响,探讨[Ca2+]I变化时Ca2+的可能来源.结果 蝎毒重组蛋白对人肺癌A549细胞生长有明显抑制作用;荧光显微镜显示,蝎毒重组蛋白作用细胞48 h后细胞内钙离子荧光强度增强,提示[Ca2+]I升高;流式细胞仪显示蝎毒重组蛋白在不同情况下可显著增加肿瘤[Ca2+]I(P<0.05).结论 蝎毒重组蛋白对人肺腺癌A549细胞增殖具有显著抑制作用,可能与升高肿瘤[Ca2+]I继而诱导肿瘤细胞凋亡有关;其升高肿瘤[Ca2+]I是通过开放肿瘤细胞膜钙通道和肿瘤细胞内钙库释放两条途径实现.     

人胎阑尾IAPP-、SS-及5-HT免疫反应性细胞的免疫组织化学研究

为了探讨胰岛淀粉样多肽，在胎阑尾中的个体发及其与其他生物活性物质的关系，用免疫组织ＰＡＰ法，对３６例１２－３８周人胎阑尾中ＩＡＰＰ免疫反应性细胞，生称抑素ＩＲ细胞５－羟色胺－ＩＲ细胞进行定位研究。结果表明，肥１２周，阑尾上皮中已可见到ＳＳ－ＩＲ细胞，而ＩＡＰＰ－及５－ＨＴ－ＩＲ细胞于１４周才见到。在整个胎期，阑尾中的ＩＡＰＰ－及ＳＳ－ＩＲ细胞始络分散存在，数量较少，而５－ＨＴ－ＩＲ细胞数量随胎龄增     

Response to a hepatitis B polypeptide vaccine in micelle form in a young adult population

Polypeptide micelles with relative molecular weights of 25,000 (p25) and 30,000 (gp30) daltons were prepared from native 22-nm hepatitis B surface antigen (HBsAg) particles. This p25/gp30 complex was alum-adsorbed, and three dosage levels (20 micrograms, 4 micrograms, and 0.8 micrograms) were administered at 0, 1, and 6 months to 51 human volunteers. Local and systemic reactions were clinically insignificant, and all vaccinees seroconverted, regardless of dose. As anticipated, antibody responses diminished as the dosage was reduced. Seroconversion rates and geometric mean antibody levels for the 20 micrograms dosage group were significantly better than those observed with a commercial vaccine and were comparable to those achieved after immunization with 40 micrograms of the intact 22-nm particles used to prepare the polypeptides. By 2 weeks, an anti-HBs response was elicited in 80% of the group receiving 20 micrograms of the polypeptide vaccine. This rapid response to immunization may be particularly beneficial for postexposure prophylaxis where the early development of immunity is advantageous.