Silkworm powder containing manganese superoxide dismutase regulated the immunity and inhibited the growth of Hepatoma 22 cell in mice

The effects of SOD contained silkworm powder on immune regulation and inhibition against Hepatoma 22 tumor cells in vivo were investigated. The activity of natural killer cell (NK) and the ConA-stimulated spleen proliferation were measured. The results found that the SOD-contained silkworm powder caused an enhancement on NK cell activity, which implied this material modulated the immune system in mice in vivo. The NK cell activities of Hepatoma 22 tumor modeled mice treated with silkworm powder including SOD were increased significantly compared to a modeled control and silkworm powder without SOD, reaching 36.18%. In addition, the ConA-stimulated spleen proliferation of SOD treated mice was higher than that of the controls. The treatment of SOD contained silkworm powder presented 40.3% of average inhibition rate to Hepatoma 22 tumor, showing stronger inhibition against tumor. There were no significant difference in body weight between modeled control and SOD silkworm powder feeding in Hepatoma 22 tumor modeled mice, suggesting the SOD silkworm powder is safety as an inhibitant to tumor. In conclusion, these findings demonstrate that administration of silkworm powder containing SOD results in activation of NK cells and immunity, suggesting the silkworm powder containing SOD plays a positive role in tumor inhibition.     

Anti-oxidation and immune responses in mice upon exposure to manganese superoxide dismutase expressed in silkworm larvae, Bombyx mori L

With manganese superoxide dismutase expressed in silkworm larvae, Bomby mori L, we investigate the effects of silkworm larvae powder containing SOD on the antioxidation and the immune system of mouse. The contents of MDA both in mice plasma or liver organ treated with silkworm larvae powder containing manganese superoxide dismutase were reduced compare to control. The superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities both in plasma or liver organ of the treated mice were significantly higher than that of both control and bromobenzene treated mice (group-BM), suggesting the silkworm larvae powder containing SOD play a positive role in anti-oxidation in mice. This experiment was also designed to investigate the effects of silkworm larvae powder containing SOD on the immune system of mouse, focused on hemolysin response, hemagglutination against SRBC and the activity of natural killer (NK) cells. All treated mice showed significant increase in hemolysin response to SRBC and demonstrated an activation of NK cell function by the SOD-contained silkworm larvae powder, which suggest a promotion in humoral immunity. The results suggested the SOD expressed in silkworm maybe have potential application in medicine.     

Immunity promotion and proteomic identification in mice upon exposure to manganese superoxide dismutase expressed in silkworm larvae

With manganese superoxide dismutase (SOD) expressed in silkworm larvae, Bomby mori L, we investigated the effects of silkworm larvae powder containing SOD on the immune system of mouse and employed a proteomics approach to examine this phenomenon. Our data on the effects of continuous treatment with SOD-containing silkworm larvae powder showed that the ConA-stimulated splenocyte proliferation of all three treated groups was higher than that of the control. The results of PFC assay also revealed that antibody production was higher in all three treated groups than controlled mice. We investigated the phagocytosis of mouse macrophages. The SOD treatment led to a dose-dependent increase of phagocytic activity. We identified six proteins that related to immunity of mice. The data showed all these six matched proteins related immunity presented the increase of expression level in plasma of mouse administrated with silkworm powder including SOD compared to that of control. These findings demonstrate that administration of silkworm larvae powder containing SOD results in enhancement of immunity activities in the mouse. The results also suggested that the SOD expressed in silkworm maybe have potential application in medicine.     

Effects of silkworm larvae powder containing manganese superoxide dismutase on immune activity of mice

To study the function of silkworm larvae powder containing superoxide dismutase and potential practical development, we investigated the safety assessment and effects on immune activity of mice such as the growth of immunity-related organs, delayed-type hypersensitivity (DTH) and charcoal particle clearance ability. The mean body weights in treated mice were significantly heavier than that of control, meanwhile, the ratio of splenocytes/body weight and the thoracic gland/body weight in treated mice was significantly enhanced after 30 days treated with silkworm larvae powder containing manganese superoxide dismutase. The treated mice resulted in a profound activation of the DTH and charcoal particle clearance, and indicated the treated mice have stronger phagocytic activity to exogenous materials. Our data also indicated the feeding treatment was safe with 360 folds of recommended human dosage in acute toxic test. In long-term test, there were no effects of silkworm larvae powder containing SOD on treated mice’s growth and inside organs as long as 90 days. Further the electronic microscope investigation showed the intestine, liver, splenocyte and stomach in mice were no obvious changes both in organs and sub-organs such as nucleus, endoplasmic reticulum, mitochondrion, Golgi and peroxisomes after treated for as long as 90 days.     

超强静磁场联合环磷酰胺对S180荷瘤小鼠抗肿瘤和相关生理指标的影响

实验探讨了超强静磁场(ultra strong static magnetic field,USMF)联合环磷酰胺(cyclophosphamide,CTX)连续处理,对S180荷瘤小鼠抗肿瘤、抗氧化、免疫及骨髓抑制等方面的影响。对S180肉瘤小鼠分组处理后,剥取肉瘤组织称重并进行病理检验。检测过氧化氢酶、超氧化物歧化酶和谷胱甘肽过氧化物酶的活力、总抗氧化能力,以及肝脏和肾脏中丙二醛的含量、脾脏和胸腺指数、脾脏淋巴细胞转化率、外周血中的白细胞数目和骨髓细胞的DNA含量。腹水瘤小鼠同样处理后正常饲养,记录生存时间。结果发现,USMF+CTX组的抑瘤率(72.5%)比CTX组(51.5%)提高了40.8%,生命延长率提高了1.5倍,抗氧化和免疫能力也有一定程度的增强。表明USMF结合CTX,可以协同性抑制S180荷瘤小鼠肿瘤的生长,并减轻CTX对小鼠的副作用。     

Superoxide dismutase induces G1‐phase cell cycle arrest by down‐regulated expression of Cdk‐2 and cyclin‐E in murine sarcoma S180 tumor cells

As an efficient reactive oxygen species–scavenging enzyme, superoxide dismutase (SOD) has been shown to inhibit tumor growth and interfere with motility and invasiveness of cancer cells. In this study, the molecular mechanisms of cell cycle arrest when S180 tumor cells were exposed to high levels of SOD were investigated. Here, both murine sarcoma S180 tumor cells and NIH‐3T3 mouse fibroblasts were respectively treated with varying concentrations of Cu/Zn‐SOD for 24, 48 and 72 h to determine optimal dose of SOD, which was a concentration of 800 U/ml SOD for 48 h. It is found that SOD induced S180 cell cycle arrest at G1‐phase with decreasing level of superoxide production, whereas SOD had less effect on proliferation of NIH‐3T3 cells. Moreover, the expression rate of Proliferating Cell Nuclear Antigen (PCNA) in S180 tumor cells was suppressed after SOD treatment, which indicated the inhibition of DNA synthesis in S180 cells. Besides, there were significant down‐regulations of cyclin‐E and Cdk‐2 in S180 cells after SOD treatment, which contributed to the blockage of G1/S transition in S180 cell cycle. Together, our data confirmed that SOD could notably inhibit proliferation of S180 tumor cell and induce cell cycle arrest at G1‐phase by down‐regulating expressions of cyclin‐E and Cdk‐2. Copyright © 2012 John Wiley & Sons, Ltd.     

软骨多糖对荷瘤小鼠免疫功能影响的初步研究

目的研究软骨多糖对荷瘤小鼠的作用,并探讨其对免疫功能的影响。方法采用小鼠肉瘤S180细胞建立动物腹水瘤模型,然后随机将小鼠分为生理盐水对照组和软骨多糖给药组,连续腹腔注射生理盐水或软骨多糖,分别测量ConA和LPS刺激下小鼠脾细胞淋巴增殖情况、外周血NK细胞的活性,及外周血单个核细胞E花环形成率。结果软骨多糖能刺激淋巴细胞增殖,明显提高NK细胞的活性,提高E花环形成率。结论软骨多糖能通过增强S180荷瘤小鼠的免疫功能而抑制肿瘤的生长。     

酯苷胶囊对小鼠移植瘤的抑制作用

目的：探讨酯苷胶囊对小鼠移植性肿瘤的抑制作用。方法：采用小鼠移植性肿瘤模型,以5-FU为阳性对照组,观测2、4、8mg·kg^-1酯苷胶囊对小鼠H22、S180肉瘤和HCA肝癌模型动物的抗肿瘤作用。结果：酯苷胶囊对H22、S180和HCA移植瘤的抑制率分别为36．8％～65．3％,19．0％～41．4％,46．8％～52．3％。结论：酯苷胶囊具有较强的抗肿瘤作用,显著延长荷瘤小鼠的生命。     

香菇C91-3菌丝发酵蛋白对S180小鼠细胞免疫功能和IFN-γ及TNF-α的作用

目的观察香菇C91-3菌丝发酵蛋白对荷瘤小鼠免疫系统的作用。方法昆明小鼠24只,S180瘤细胞荷瘤建立动物模型,随机平均分为3组：香菇C91-3菌丝发酵蛋白治疗组、环磷酰胺治疗组、生理盐水对照组。观察香菇C91-3菌丝发酵蛋白对荷瘤小鼠NK细胞活性、淋巴细胞转化率、IFN-γ和TNF-α等免疫指标的影响。结果香菇C91-3菌丝发酵蛋白能够显著提高治疗组小鼠的NK细胞活性、淋巴细胞转化率和血清中IFN-γ和TNF-α的含量。结论香菇C91-3菌丝发酵蛋白能够显著提高荷瘤小鼠免疫系统的活性。     

灵芝发酵液多糖提取物对荷瘤小鼠细胞免疫的动态观察

目的:观察灵芝发酵液多糖提取物对S180荷瘤小鼠部分免疫指标的动态调节作用,探讨其抗肿瘤机制.方法:S180瘤细胞荷瘤昆明小鼠80只建立动物模型,生理盐水组(NS组)与灵芝发酵液多糖组(GFG组)各40只,分别于荷瘤后第4,7,10,13,16天每组各处死8只小鼠,检测GFG对NK细胞活性、淋巴细胞转化率的影响.结果:GFG能显著提高荷瘤小鼠NK细胞活性和淋巴细胞转化率.随荷瘤时间延长,GFG组较NS组能维持较高水平(P＜0.01),但总体呈下降趋势.结论:灵芝发酵液多糖提取物能显著提高NK细胞活性和淋巴细胞转化率,并维持一定水平.     

软骨多糖诱导小鼠S180细胞凋亡的作用机制

目的研究软骨多糖对S180荷瘤小鼠的作用,并探讨其抑瘤作用机制。方法采用小鼠肉瘤S180细胞建立动物腹水瘤模型,通过腹腔注射软骨多糖进行治疗,治疗期间抽取腹水瘤细胞进行细胞生物学分析。通过HE染色,流式细胞术、TUNEL法检测细胞形态学方面、细胞周期及凋亡率的变化情况;通过免疫荧光方法检测Fas、增殖细胞核抗原(PCNA)的表达情况。结果软骨多糖可以明显提高S180荷瘤小鼠的生存率,细胞形态学观察可见细胞出现细胞质浓缩、核固缩及凋亡小体等现象。软骨多糖作用后的S180细胞,其细胞周期被阻遏于G2/M期,Fas蛋白的表达水平于给药24 h后升高,增殖细胞核抗原PCNA表达下降。结论软骨多糖可能通过影响肿瘤细胞周期和Fas、PCNA蛋白的表达来诱导S180细胞凋亡,并显著抑制肿瘤细胞的生长,延长S180荷瘤小鼠的生存时间,研究证实动物软骨多糖具有潜在的药用价值。     

盐酸洛拉曲克在体内、外对S-180细胞株的抗增殖作用

研究合合成新型胸苷合成酶抑制剂盐酸洛拉曲克在体内、外对S－180细胞株及正常人胚肾HEK293细胞的抗增殖作用;使用MTT法测定抑制率,以提高荷腹水瘤小鼠存活时间及荷实体瘤瘤重减轻情况为指标考察盐酸洛拉曲克对S－180所致肿瘤的治疗作用。结果表明：在体外盐酸洛拉曲克对S－180肿瘤细胞株有较强的细胞毒作用,对正常细胞HEK293抑制作用较弱（P＜0.05）;体内实验显示盐酸洛拉曲克可明显提高荷腹水瘤小鼠的存活时间,减轻荷实体瘤小鼠的瘤重,疗效与氟尿嘧啶（10mg/kg）相当（P＞0.05）或更好（P＜0.05）。可见,盐酸洛拉曲克在体内、外对S－180肿瘤细胞有显著的抗增殖作用,在体外对肿瘤细胞 具有选择性的抗增殖作用。     

Immune recognition of tumor cells in mice infected with Pichinde virus

Summary Pichinde virus (PV), a member of the Arenaviridae family, protects mice from a lethal inoculation with the sarcoma 180 (S180) tumor cell line. Virus replication, which is required for protection, occurs primarily in the spleen and tumor. During the first 4 days, elevated natural killer (NK) cell activity parallels an increase in serum interferon in PV-infected mice. On day 7 after infection virus-specific cytotoxic T cells (CTLs) are found in the mouse. This strong response peaks on day 13 and gradually declines over the next 17 days. The tumor-specific CTL response appears more slowly and is less intense than the virus-specific response, especially in the uninfected mouse. However, CTLs from either type of mouse recognize PV-infected tissue culture S180 target cells better than uninfected ones. Even though the primary tumor-specific immune response appears weak, mice that have cleared both virus and tumor are refractory to a subsequent challenge with S180 cells and rapidly produce tumor-specific CTLs. Thus, our data indicate a number of ways in which virus infection could lead to immune elimination of tumors: (1) Virus-induced interferon stimulates NK-cell activity, which in turn could control tumor load until a specific response is mounted against the S180 cells; (2) early onset of the tumor-specific T-cell response could be brought about by viral-enhanced tumor antigen presentation to the immune system; and (3) the tumor-specific T-cell response could be augmented through a bystander phenomenon involving factors associated with T cells responding specifically and vigorously to the virus itself.Deceased     

The anti-tumor activity of Mikania micrantha aqueous extract in vitro and in vivo

Aqueous extract obtained from Mikania micrantha (MMAE) is commonly used as traditional medicine in some countries. We hypothesized that MMAE may inhibit tumor cell growth, both in an in vitro and in vivo setting. In in vitro experiments, two kinds of human cancer cell lines, K562 and Hela were used to test the anti-tumor activity. Inhibitory concentrations (IC₅₀) were obtained from the inhibition curves fitted by regression analysis, inhibitory rates (%) were calculated by MTT assay, morphological changes were observed by transmission electron microscope (TEM), cell cycles were analyzed by flow cytometry (FCM), and DNA ladders were determined by agarose gel electrophoresis. The in vivo anti-tumor activity was evaluated by calculating the tumor inhibitory rates, thymus index and spleen index of S180-bearing mice. Paraffin-embedded sections were used to test the pathologic changes. The result displayed that the growth of K562 and Hela were enhanced when treated with MMAE at 20 μg/mL after 48 h. Other concentrations of MMAE (50, 100, 200, 400 μg/mL) inhibited the proliferation of both kinds of cells. The IC₅₀ values of K562 and Hela at 48 h were 167.16 and 196.27 μg/mL and at 72 h 98.07 and 131.56 μg/mL, respectively. The effects showed time-dose dependence. MMAE led to damages of organelles and induced apoptosis. These results were confirmed by ladder DNA fragmentation profile. MMAE also increased the percentage of cells in G2/M phase and decreased the percentage of cells undergoing G0/G1 and S phase in in vivo tests using S180 cells. MMAE showed antitummor activity in vivo, with its tumor inhibitory rate ranging from 12.1 to 46.9 %. MMAE also induced necrosis, as shown by pathological examination of Hematoxilin-Eosin stained tumor sections. Meanwhile, compared with the control group, the changes of thymus index and spleen index in MMAE treated group were not obvious. This study suggests that MMAE may be an effective agent for cancer therapy with low toxicity.     

婴儿双歧杆菌对小鼠肉瘤S180的抑制作用

本文主要观察婴儿双歧杆菌(Bif. 189—3)对小鼠皮下移植肉瘤180(S180)的抑制作用。结果发现,该菌无论在S180移植前或移植后经皮下注射,均能明显抑制皮下移植S180的增长,并使带瘤鼠存活期显著延长。病理检查见实验组肿瘤坏死明显,肿瘤周围有大量炎症细胞浸润。提示该菌能非特异性增强肿瘤局部的免疫反应。     

C_(60)对小鼠S_(180)肉瘤光动力学作用模型的建立

为验证Ｃ６０对活体肿瘤的光动力学损伤作用,我们从两方面进行实验：Ｃ６０对荷瘤小鼠的Ｓ１８０实体瘤的光动力学杀伤作用和Ｃ８０对离体Ｓ１８０肿瘤细胞的光动力学杀伤作用,在小鼠的瘤体上注射Ｃ８０光敏剂,在５１１ｎｍ、５７８ｎｍ混合黄录色激光照射正派主发Ｃ６０,产生大量的单线态氧,杀伤活性肿瘤。激光光强为５００ｍＷ,Ｃ６０浓度为３０μｇ／ｍｌ时,荷瘤小鼠寿命平均延长５天,瘤径减小１ｃｍ,瘤重减轻０．８克,     

Immunomodulatory Activity of 3β,6β‐Dihydroxyolean‐12‐en‐27‐oic Acid in Tumor‐Bearing Mice

3β,6β‐Dihydroxyolean‐12‐en‐27‐oic acid ( 1 ) is a pentacyclic triterpenoid isolated from the rhizomes of Astilbe chinensis. To evaluate the in vivo antitumor potential and to elucidate its immunological mechanisms, effect of 1 on the growth of mouse‐transplantable tumors, and the immune response in naive and tumor‐bearing mice were investigated. The mice inoculated with mouse tumor cell lines were orally treated with 1 at the doses of 40, 60, and 80 mg/kg for 10 days. The effects of 1 on the growth of mouse‐transplantable S180 sarcoma and H22 hepatoma, splenocyte proliferation, cytotoxic T lymphocyte (CTL) activity, natural killer (NK) cell activity, and production of interleukin‐2 (IL‐2) from splenocytes in S180‐bearing mice were measured. Furthermore, the effect of 1 on 2,4‐dinitrofluorobenzene (DNFB)‐induced delayed‐type hypersensitivity (DTH) reactions and the sheep red blood cell (SRBC)‐induced antibody response in naive mice were also studied. Compound 1 could not only significantly inhibit the growth of mouse transplantable S180 sarcoma and H22 hepatoma, increase splenocytes proliferation, CTL and NK cell activity, and the level of IL‐2 secreted by splenocytes in tumor‐bearing mice, but also remarkably promote the DTH reaction and enhance anti‐SRBC antibody titers in naive mice. These results suggested that 1 could improve both cellular and humoral immune response, and could act as antitumor agent with immunomodulatory activity.     

金黄色葡萄球菌肠毒素A(SEA)对小鼠体内肉瘤抑制效果

以实验室建立的$180小鼠肿瘤模型为研究对象,采用腹腔注射给药,观察葡萄球菌肠毒素A（SEA）在体内抑制肿瘤的效果。实验表明,SEA抑肿瘤率为40．18％,显示对肿瘤有一定的抑制作用;能显著刺激脾脏细胞增殖,使脾指数升高至11．3mg/g;使血清和脾组织中IL-2水平分别升至69．77pg/mL和682．43pg/mL;且能诱导肿瘤组织中产生大量的CD4^＋T细胞和CD8^＋T细胞。结果显示,SEA在机体内对免疫功能有正向调节作用,从而在一定程度上抑制了肿瘤细胞的生长。     

Curcumin reverses breast tumor exosomes mediated immune suppression of NK cell tumor cytotoxicity

An important characteristic of tumors is that they at some point in their development overcome the surveillance of the immune system. Tumors secrete exosomes, multivesicular bodies containing a distinct set of proteins that can fuse with cells of the circulating immune system. Purified exosomes from TS/A breast cancer cells, but not non-exosomal fractions, inhibit (at concentrations of nanograms per ml protein) IL-2-induced natural killer (NK) cell cytotoxicity. The dietary polyphenol, curcumin (diferuloylmethane), partially reverses tumor exosome-mediated inhibition of natural killer cell activation, which is mediated through the impairment of the ubiquitin-proteasome system. Exposure of mouse breast tumor cells to curcumin causes a dose-dependent increase in ubiquitinated exosomal proteins compared to those in untreated TS/A breast tumor cells. Furthermore, exosomes isolated from tumor cells pretreated with curcumin have a much attenuated inhibition of IL-2 stimulated NK cell activation. Jak3-mediated activation of Stat5 is required for tumor cytotoxicity of IL-2 stimulated NK cells. TS/A tumor exosomes strongly inhibit activation of Stat5, whereas the tumor exosomes isolated from curcumin-pretreated tumor cells have a lowered potency for inhibition of IL-2 stimulated NK cell cytotoxicity. These data suggest that partial reversal of tumor exosome-mediated inhibition of NK cell tumor cytotoxicity may account for the anti-cancer properties of curcumin.     

小鼠S_(180)瘤细胞免疫性的初步研究

本文选用灵敏度高,特异性强的酶标免疫吸附测定法(ELISA)及A.D.C.C.测试法(Antibody Dependent Cell mediated Cytotoxicity Assay)检测到以S_(180)瘤细胞免疫的小鼠血清中有抗S_(180)瘤细胞的特异抗体。并初步探讨了S_(180)肿瘤细胞膜上的唾液酸与抗原抗体相互作用的关系。实验表明S_(180)肿瘤细胞经唾液酸酶作用,水解去除瘤细胞表面的唾液酸后,对其与特异抗体的结合反应无明显影响。这提示S_(180)瘤细胞膜表面的糖蛋白上的唾液酸未必直接参与抗原抗体的特异性结合。以上结果为进一步研究患瘤小鼠各阶段的免疫情况及寻求最佳免疫时机,为治疗肿瘤奠定了基础。