大黄素阻抑人乳腺癌MCF-7细胞增殖和细胞周期进程

 目的：探讨大黄素抗人乳腺癌MCF-7细胞增殖及其相关机制。方法：采用MTT法检测大黄素对MCF-7细胞的增殖抑制作用;流式细胞术分析周期分布和凋亡情况;原子力显微镜(AFM)观察细胞膜表面超微结构的变化。结果：大黄素呈剂量依赖性地抑制MCF-7细胞的增殖;大黄素能使MCF-7细胞阻滞在G0/G1期;Annexin V/PI双染法结果表明,大黄素对MCF-7细胞没有明显的促凋亡作用。AFM观察细胞膜超微结构,结果显示对照组细胞核区饱满,膜表面平坦光滑;大黄素作用48 h可致细胞核区坍塌萎缩,细胞表面颗粒密集,膜表面的平均粗糙度(Ra) 和均方粗糙度(Rq) 与对照组相比均有显著增加(P<005)。结论: 大黄素通过阻断MCF-7细胞的细胞周期进程及影响细胞膜超微结构而发挥抗肿瘤作用。     

Interleukin-16 aggravates ovalbumin-induced allergic inflammation by enhancing Th2 and Th17 cytokine production in a mouse model

Asthma is a chronic inflammatory disease that involves a variety of cytokines and cells. Interleukin-16 (IL-16) is highly expressed during allergic airway inflammation and is involved in its development. However, its specific mechanism of action remains unclear. In the present study, we used an animal model of ovalbumin (OVA)-induced allergic asthma with mice harboring an IL-16 gene deletion to investigate the role of this cytokine in asthma, in addition to its underlying mechanism. Increased IL-16 expression was observed during OVA-induced asthma in C57BL/6J mice. However, when OVA was used to induce asthma in IL-16^−/− mice, a diminished inflammatory reaction, decreased bronchoalveolar lavage fluid (BALF) eosinophil numbers, and the suppression of OVA-specific IgE levels in the serum and BALF were observed. The results also demonstrated decreased levels of T helper type 2 (Th2) and Th17 cytokines upon OVA-induced asthma in IL-16^−/− mice. Hence, we confirmed that IL-16 enhances the lung allergic inflammatory response and suggest a mechanism possibly associated with the up-regulation of IgE and the promotion of Th2 and Th17 cytokine production. This work explored the mechanism underlying the regulation of IL-16 in asthma and provides a new target for the clinical treatment of asthma.     

Taurine chloramine inhibits lymphocyte proliferation and decreases cytokine production in activated human leukocytes

Previously, we described the inhibition of proinflammatory mediators such as nitric oxide, tumor necrosis factor-alpha (TNF-alpha), and prostaglandin E2 by taurine chloramine (Tau-Cl) in activated rodent macrophages. We also demonstrated that Tau-Cl suppressed superoxide anion, IL-6, and IL-8 production in activated human polymorphonuclear leukocytes separated from peripheral blood. In these studies, we report the effect of Tau-Cl on lymphocyte proliferation and the production of cytokines by activated human peripheral blood mononuclear leukocytes. Adherent and nonadherent leukocytes were activated using lipopolysaccharide (LPS) and phytohemagglutinin (PHA), respectively, in the presence or absence of Tau-Cl. Tau-Cl significantly suppressed lymphocyte proliferation as measured by tritiated (3H) thymidine. Production of IL-6, IL-8, and IL-2 in PHA-activated nonadherent leukocytes was inhibited by Tau-Cl. The production of IL-1beta, IL-6, and IL-8 was also decreased in LPS-activated adherent monocytes by Tau-Cl. These data demonstrate that the ability of Tau-Cl to modulate the immune response is not species specific and extends to human leukocytes.     

UVA1 irradiation inhibits fibroblast proliferation and alleviates pathological changes of scleroderma in a mouse model

The purpose of the present study was to compare the effects of different doses of ultraviolet radiation A1 (UVA1) on human fibroblast proliferation and collagen level in a mouse model of scleroderma,so as to identify appropriate irradiation doses for clinical treatment of scleroderma.Monolayer from human fibroblasts was cultured in vitro,and a mouse model of scleroderma was established by subcutaneous injection of 100 μL of 400 μg/mL bleomycin into the back of BALB/c mice for 4 weeks.The mouse models and human fibroblasts were divided into UVA1exposed (100,60 and 20 J/cm 2) and UVA-unexposed groups.At 0,24 and 48 h after exposure,cell proliferation and levels of hydroxyproline and collagen were detected.UVA1 irradiation was performed 3 times weekly for 10 weeks,and the pathological changes of skin tissues,skin thickness and collagen level were observed after phototherapy.Cell proliferation and the levels of hydroxyproline and collagen were inhibited after phototherapy,and there was a significant difference between the UVA1-exposed cells and UVA1-unexposed cells (P < 0.001).In addition,UVA1 phototherapy improved dermal sclerosis and softened the skin,and there were significant differences between the high-dose UVA1 group and the model group,and the negative group (P < 0.05).It is concluded that UVA1 radiation can reduce cell proliferation,and decrease hydroxyproline and collagen levels in a dose-dependent manner in vitro.High-dose UVA1 phototherapy has marked therapeutic effect on scleroderma in the mouse model.Decreased collagen level may be related to the reduced number and activity of cells,as well as inhibition of collagen synthesis.     

Dihydroartemisinin suppresses ovalbumin-induced airway inflammation in a mouse allergic asthma model

Abstract

Asthma is a complex disease characterized by reversible airway obstruction, airway hyper-responsiveness (AHR) and chronic inflammation of the airways. Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin isolated from the traditional Chinese herb Artemisia annua, has been shown to possess antimalarial and antitumor activities, but whether it can be used in asthma treatment has not been investigated. In this study, we attempted to determine whether DHA regulates inflammatory mediators in the ovalbumin (OVA)-induced mouse asthma model. BALB/c mice were sensitized and challenged by OVA to induce chronic airway inflammation. The intragastrical administration of DHA at 30?mg/kg significantly decreased the number of infiltrating inflammatory cells, T-helper type 2 (Th2) cytokines, OVA-specific immunoglobulin E (IgE) and AHR. Treatment with DHA also attenuated OVA-induced mRNA expression of Muc5ac and chitinase 3-like protein 4 (Ym2) in lung tissues. In addition, lung histopathological studies revealed that DHA inhibited inflammatory cell infiltration and mucus hypersecretion. Then signal transduction studies showed that DHA significantly inhibited extracellular signal-regulated protein kinase (ERK), p38 mitogen-activated protein kinase phosphorylation. DHA also inhibited nuclear factor-κB (NF-κB) activation via the inhibition of phosphorylation of IκBα. These findings provide new insight into the immunopharmacological role of DHA in terms of its effects in a mouse model of asthma.     

Supplementation with vitamins C, E, β-carotene and selenium has no effect on anti-oxidant status and immune responses in allergic adults: a randomized controlled trial

BACKGROUND: Anti-oxidants are of growing interest in early treatment and prevention of allergic diseases in early life, but the effects on allergen-specific immune responses need to be documented further before intervention studies in infants are undertaken. The aim of this study in adults was to determine the effects of dietary anti-oxidants on allergen-specific immune responses in sensitized individuals. METHODS: In a randomized controlled trial, 54 allergic adults received an anti-oxidant supplement (n=36) comprising beta-carotene (9 mg/day), vitamin C (1500 mg/day), vitamin E (130 mg/day), zinc (45 mg/day), selenium (76 microg/day) and garlic (150 mg/day) or a placebo (n=18) for 4 weeks. Anti-oxidant capacity (AC), serum levels of vitamin C, vitamin E, beta-carotene and selenium, peripheral blood responses, exhaled nitric oxide (eNO), as a marker of airway inflammation, and plasma F(2) isoprostanes, as a measure of oxidative stress, were measured before and after supplementation. RESULTS: Anti-oxidant supplementation resulted in significant increases in serum levels of vitamin C, vitamin E, beta-carotene and selenium levels, compared with the placebo group (P<0.001). There was no change in serum AC, plasma F(2)-isoprostanes, eNO or immune responses following supplementation with anti-oxidants compared with placebo. CONCLUSION: Supplementation with anti-oxidants resulted in significantly increased levels of vitamin C, vitamin E, beta-carotene and selenium but no change in immune responses, serum AC or plasma F(2)-isoprostanes.     

小鼠晶状体发育过程中细胞增殖及突触素蛋白的表达

目的 探讨小鼠晶状体细胞增殖、细胞周期蛋白（cyclin) D1和突触素（SYN）的表达情况。方法 各日龄小鼠共150只,HE染色和
4’6-二脒基-2-苯基吲哚（DAPI）染色,在光镜下观察小鼠晶状体的一般结构;用抗细胞增殖核抗原（PCNA）免疫荧光法以及5′-溴脱氧尿嘧
啶核苷（BrdU）技术标记晶状体增殖细胞;用免疫荧标记标价法检测细胞周期蛋白cyclin D1和SYN在晶状体的表达。结果 孕8d（E8）左右,
晶状体由最初的晶状体板分化而来,是由单层柱状上皮细胞围成的空泡,空泡再进一步分化,被原始晶状体纤维所替代,逐渐形成晶状体;
BrdU和PCNA的阳性细胞主要分布在晶状体上皮细胞的前囊中央区,但出生10d（P10）以后BrdU阳性细胞已不在晶状体上皮细胞分布,而PCNA持
续表达。Cyclin D1阳性细胞在胚胎期主要在晶状体上皮细胞的赤道部表达,出生早期在晶状体上皮细胞的前囊中央区,出生5d（P5）以后在赤
道前区分布,赤道部有少量阳性细胞;在晶状体发育过程中,SYN可以在晶状体基质中和赤道部胞体中发现。结论 细胞增殖对晶状体的发育和
修复起着重要的作用;cyclin D1可能参与调控赤道部细胞的消失,因此保持晶状体的透明度;SYN在晶状体的表达说明了它对晶状体的透明有
着重要的作用,通过调控钙离子内流入胞体和基质中。     

唑来膦酸抑制U937细胞增殖及诱导凋亡

目的:研究唑来膦酸(ZOL)对人类急性髓系白血病细胞U937的增殖抑制及促凋亡作用。方法:CCK-8法检测不同时间ZOL对U937细胞的生长抑制率;流式细胞术检测ZOL对U937细胞周期的影响;Annexin V-PI法及Hoechst 33342法检测ZOL作用前后细胞凋亡情况变化,JC-1检测ZOL对U937细胞线粒体膜电位变化的影响;克隆形成实验检测U937细胞克隆形成能力;Western blot法检测ZOL对U937细胞周期和凋亡相关蛋白的变化。结果:CCK-8结果显示ZOL可以抑制U937细胞的活力,并呈时间-剂量依赖性;Annexin V-PI及Hoechst33342结果显示ZOL可以促进U937细胞凋亡,且呈时间-剂量依赖性;JC-1结果显示ZOL可以明显降低U937细胞线粒体膜电位;PI法证实ZOL将U937细胞周期阻滞在S期,克隆形成实验证实0.2 mmol/L ZOL可以完全抑制U937细胞的克隆形成能力;Western blot结果显示ZOL作用于U937细胞48 h后细胞周期相关蛋白p21表达显著增强,促凋亡蛋白Bax表达增强,抑凋亡蛋白Bcl-2表达明显减弱。结论:ZOL抑制U937细胞的增殖和克隆形成主要是由于抑制了细胞周期相关蛋白表达,同时ZOL可以促进U937细胞凋亡,这种作用主要是通过调节线粒体凋亡途径相关蛋白来实现的。     

Cannabinoids attenuate cancer pain and proliferation in a mouse model

We investigated the effects of cannabinoid receptor agonists on (1) oral cancer cell viability in vitro and (2) oral cancer pain and tumor growth in a mouse cancer model. We utilized immunohistochemistry and Western blot to show that human oral cancer cells express CBr1 and CBr2. When treated with WIN55,212-2 (non-selective), ACEA (CBr1-selective) or AM1241 (CBr2-selective) agonists in vitro, oral cancer cell proliferation was significantly attenuated in a dose-dependent manner. In vivo, systemic administration (0.013 M) of WIN55,212-2, ACEA, or AM1241 significantly attenuated cancer-induced mechanical allodynia. Tumor growth was also significantly attenuated with systemic AM1241 administration. Our findings suggest a direct role for cannabinoid mechanisms in oral cancer pain and proliferation. The systemic administration of cannabinoid receptor agonists may have important therapeutic implications wherein cannabinoid receptor agonists may reduce morbidity and mortality of oral cancer.     

Chloroquine inhibits T cell proliferation by interfering with IL-2 production and responsiveness.

Chloroquine (Chl) is an anti-rheumatic drug that is widely used in the treatment of rheumatoid arthritis (RA). It seems that T cells are important in the pathogenesis of RA, but it is not known whether Chl acts via inhibition of T cell function. We here present evidence that Chl, just like cyclosporine A (CsA), inhibits T cell proliferation as induced with immobilized alpha CD3 MoAb in a concentration-dependent manner, at least partly through interfering with the production of IL-2 protein and the induction of IL-2 mRNA. Furthermore, Chl impedes the responsiveness of T cell clones to IL-2 since (1) the inhibition of alpha CD3 MoAb-induced proliferation by Chl could not be reversed by rIL-2 and (2) Chl directly blocks IL-2-driven proliferation of cloned T cells. Chl appeared to interfere with the internalization (50% inhibition) and degradation (total blockade) of rIL-2. Finally, the combination of Chl and CsA synergistically inhibited T cell proliferation. We conclude that Chl may inhibit functional properties of human T cells, although the drug is 100- to 1000-fold less potent than CsA in inhibiting T cell proliferation and IL-2 production, respectively. It is speculated that the in vitro effects of Chl might be relevant in explaining the anti-rheumatic effect of this drug in patients with RA.     

alpha-Lipoic acid inhibits airway inflammation and hyperresponsiveness in a mouse model of asthma

BACKGROUND: Oxidative stress may play an important role in the pathogenesis of bronchial asthma. OBJECTIVE: We evaluated the therapeutic effect of alpha-lipoic acid, a nonenzymatic antioxidant, in a mouse model of asthma. METHODS: BALB/c mice were immunized intraperitoneally with ovalbumin (OVA) on days 1 and 14 and challenged with inhaled OVA on days 28, 29, and 30. Mice were fed OVA-free standard mouse chow with 0%, 0.125%, 0.25%, 0.5%, and 1% (wt/wt) alpha-lipoic acid during the immunization and challenge periods. On day 31, mice were challenged with inhaled methacholine, and enhanced pause was measured as an index of airway hyperresponsiveness. Severity of airway inflammation was determined by means of differential cell count of bronchoalveolar lavage (BAL) fluid and by means of histopathologic lung analysis. Levels of OVA-specific IgE in serum, IL-4 and IL-5 in BAL fluid, and intracellular reactive oxygen species in alveolar macrophages and lymphocytes obtained from regional perihilar lymph nodes were measured. Nuclear factor kappaB DNA-binding activity in lung tissues was analyzed by means of electrophoretic gel mobility shift assay. RESULTS: Compared with untreated asthmatic mice, mice treated with alpha-lipoic acid had significantly reduced airway hyperresponsiveness, a lower proportion of eosinophils among BAL cells, and significantly improved pathologic lesion scores of the lungs. alpha-Lipoic acid also significantly reduced serum OVA-specific IgE concentrations, IL-4 and IL-5 concentrations in BAL fluid, and intracellular reactive oxygen species and nuclear factor kappaB DNA-binding activity. CONCLUSION: These results suggest that oxidative stress plays an important role in asthmatic airway inflammation and that alpha-lipoic acid may be useful as adjuvant therapy for bronchial asthma.     

Airway cytokine responses to acute and repeated stress in a murine model of allergic asthma

Although stress is known to exacerbate asthma, the underlying mechanisms are unclear. Using a murine model of allergic asthma, Th1 (interleukin [IL]-2 and gamma-interferon [IFN-γ]) and Th2 (IL-4 and IL-5) airway cytokine responses and Th1:Th2 cytokine ratios to acute and repeated stress were examined in bronchoalveolar lavage fluid (BALF). Asthmatic mice showed significantly higher IL-4 level and Th2 predominance, compared with healthy mice. To acute stress, asthmatic mice significantly increased IL-4 but decreased IFN-γ levels favoring Th2 predominance, whereas healthy mice significantly decreased IL-4 level favoring Th1 predominance in BALF. To repeated stress, asthmatic mice significantly decreased IFN-γ, IL-4 and IL-5 levels overall favoring Th1 predominance, whereas healthy mice basically maintained the same response profile shown to acute stress. These findings suggest that a significant shift toward Th2 predominance in asthmatic mice under acute stress may be a mechanism underlying exacerbation of asthma.     

丹参酮ⅡA对C6胶质瘤细胞的增殖抑制和诱导凋亡作用

目的：探讨丹参酮ⅡA对大鼠胶质瘤细胞株C6的增殖抑制作用及其作用机理。 方法： 应用MTT比色法检测不同浓度丹参酮ⅡA对C6细胞的增殖抑制，应用流式细胞仪观察丹参酮ⅡA对C6细胞周期的影响，应用琼脂糖凝胶电泳观察C6细胞DNA的变化，应用RT-PCR观察相关基因c-myc表达的变化。 结果： 丹参酮ⅡA对C6细胞增殖抑制作用呈剂量依赖性。细胞周期分析显示：用1.0 mg/L丹参酮ⅡA作用C6细胞，在作用72 h出现凋亡峰，凋亡细胞占细胞总数7.7%；用2.0 mg/L丹参酮ⅡA作用C6细胞，在作用72 h出现凋亡峰，凋亡细胞占细胞总数21.6%。琼脂糖凝胶电泳结果显示：一定剂量丹参酮ⅡA作用后，C6细胞具有明显的凋亡特征，细胞核DNA呈梯状降解。RT-PCR结果显示：随着丹参酮ⅡA作用剂量的增加，c-myc基因的表达被明显抑制。 结论： 丹参酮ⅡA对大鼠胶质瘤细胞系C6的增殖具有明显的抑制作用及诱导凋亡的作用，并对原癌基因c-myc的表达具有抑制作用。     

Short-term simvastatin treatment has no effect on plasma cytokine response in a human in vivo model of low-grade inflammation

Statins reduce plasma cholesterol, but clinical trials and in vitro studies indicate that they might also possess anti-inflammatory properties. The effect of simvastatin on circulating cytokines and leucocytes was evaluated in a human in vivo model of low-grade inflammation. Thirty young healthy male participants received an injection of the bacterial cell wall product endotoxin (0.06 ng/kg) to induce systemic inflammation. Participants were then randomized into a control and a simvastatin group. The simvastatin group received simvastatin 20 mg daily for 14 days. All participants returned after 14 days to receive a second endotoxin injection. Plasma concentrations of tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-1 receptor antagonist were measured by enzyme-linked immunosorbent assay (ELISA) before and hourly for 6 hours after endotoxin administration. Plasma cytokines as well as total leucocyte counts increased in all participants upon endotoxin challenge but were not affected by simvastatin treatment. Tolerance to endotoxin was observed in both groups after 14 days. Short-term treatment with simvastatin (20 mg/day) did not influence circulating cytokine levels during endotoxaemia in this human in vivo study.     

三七提取物对小鼠淋巴细胞体外增殖和细胞周期的影响

目的 分析三七提取物(SE)对小鼠淋巴细胞体外增殖和细胞周期的影响,探讨其免疫抑制作用机制.方法 分离小鼠淋巴结细胞,以CFSE染色,加入多克隆刺激剂刀豆蛋白(Con A)或者佛波醇酯(PDB)和离子霉素(Ion)进行刺激,以流式细胞仪分析三七提取物对淋巴细胞增殖的影响;用碘化丙锭(PI)染色分析细胞周期分布.结果 CFSE染色分析显示,SE对Con A或者佛波醇酯(PDB)和离子霉素(Ion)诱导的小鼠淋巴细胞增殖,具有明显的抑制作用(P＜0.05),且呈剂量依赖性.对细胞周期分析表明,随着三七提取物浓度的增加,均能明显阻止淋巴细胞进入S期和G2/M期,而处于亚二倍体峰的细胞数基本不变.结论 三七提取物对小鼠淋巴细胞的增殖有明显抑制作用,它能抑制淋巴细胞进入细胞分裂周期,这种抑制作用表现出明显的周期特异性.     

Rosmarinic acid in perilla extract inhibits allergic inflammation induced by mite allergen, in a mouse model

BACKGROUND: Perilla and its constituent rosmarinic acid have been suggested to have anti-allergic activity. However, few studies have examined the effects on allergic asthma. OBJECTIVE: The purpose of this study was to evaluate the effect of oral administration of perilla leaf extract, which contains high amount of rosmarinic acid, on a murine model of allergic asthma induced by house dust mite allergen. METHODS: C3H/He mice were sensitized by intratracheal administration of Dermatophagoides farinae (Der f). Mice were orally treated with rosmarinic acid in perilla extract (PE) (1.5 mg/mouse/day). RESULTS: Der f challenge of sensitized mice elicited pulmonary eosinophilic inflammation, accompanied by an increase in lung expression of IL-4 and IL-5, and eotaxin. Daily treatment with rosmarinic acid in PE significantly prevented the increases in the numbers of eosinophils in bronchoalveolar lavage fluids and also in those around murine airways. Rosmarinic acid in PE treatment also inhibited the enhanced protein expression of IL-4 and IL-5, and eotaxin in the lungs of sensitized mice. Der f challenge also enhanced allergen-specific IgG1, which were also inhibited by rosmarinic acid in PE. CONCLUSION: These results suggest that oral administration of perilla-derived rosmarinic acid is an effective intervention for allergic asthma, possibly through the amelioration of increases in cytokines, chemokines, and allergen-specific antibody.     

小檗碱对小鼠淋巴细胞体外增殖和细胞周期的影响

目的 :分析小檗碱 (Ber)对小鼠淋巴细胞体外增殖和细胞周期的影响 ,探讨其免疫抑制作用的机制。方法 :分离小鼠淋巴结细胞 ,以CFSE染色 ,加入多克隆刺激剂刀豆蛋白 (ConA)或者佛波醇酯 (PDB)和离子霉素 (Ion)进行刺激 ,以流式细胞仪分析小檗碱对淋巴细胞增殖率的影响 ;以WST 1检测淋巴细胞的增殖情况 ;用碘化丙锭 (PI)染色分析细胞周期分布 ;细胞凋亡以Annexin V染色进行分析。结果 :CFSE染色分析显示 ,当浓度为 10 0、30和 10 μmol L时 ,Ber无论对ConA或PDB Ion诱导的小鼠淋巴细胞增殖 ,都具有明显的抑制作用 (P <0 0 5 )。以WST 1分析细胞增殖情况 ,也获得相似的结果。对细胞周期分析表明 ,随着小檗碱浓度的增加 ,处于亚二倍体峰的细胞数增加 ,处于S和G2 M期的细胞数则减少 ,而G0 G1 期的细胞数基本不变。Annexin V染色法结果表明 ,30 μmol LBer组Annexin V单阳性区域的百分率为 ( 7 13± 1 0 8) % ,与PDB Ion阴性组的百分率 ( 2 4 9± 0 2 5 ) %比较有显著的差异 (P <0 0 1)。结论 :小檗碱对小鼠淋巴细胞的增殖有明显抑制作用 ,它能抑制淋巴细胞进入细胞分裂周期 ,这种抑制作用表现出明显的周期特异性。同时 ,Ber可以诱导体外培养的小鼠淋巴细胞发生凋亡。