Anti-inflammatory Effects of Ethanolic Extract from Sargassum horneri (Turner) C. Agardh on Lipopolysaccharide-Stimulated Macrophage Activation via NF-κB Pathway Regulation

Inflammation is major symptom of the innate immune response by infection of microbes. Macrophages, one of immune response related cells, play a role in inflammatory response. Recent studies reported that various natural products can regulate the activation of immune cells such as macrophage. Sargassum horneri (Turner) C. Agardh is one of brown algae. Recently, various seaweeds including brown algae have antioxidant and anti-inflammatory effects. However, anti-inflammatory effects of Sargassum horneri (Turner) C. Agardh are still unknown. In this study, we investigated anti-inflammatory effects of ethanolic extract of Sargassum horneri (Turner) C. Agardh (ESH) on RAW 264.7 murine macrophage cell line. The ESH was extracted from dried Sargassum horneri (Turner) C. Agardh with 70% ethanol and then lyophilized at ?40?°C. ESH was not cytotoxic to RAW 264.7, and nitric oxide (NO) production induced by LPS-stimulated macrophage activation was significantly decreased by the addition of 200?μg/mL of ESH. Moreover, ESH treatment reduced mRNA level of cytokines, including IL-1β, and pro-inflammatory genes such as iNOS and COX-2 in LPS-stimulated macrophage activation in a dose-dependent manner. ESH was found to elicit anti-inflammatory effects by inhibiting ERK, p-p38 and NF-κB phosphorylation. In addition, ESH inhibited the release of IL-1β in LPS-stimulated macrophages. These results suggest that ESH elicits anti-inflammatory effects on LPS-stimulated macrophage activation via the inhibition of ERK, p-p38, NF-κB, and pro-inflammatory gene expression.     

羊栖菜多糖含量测定及多糖组分分析

目的：提取羊栖菜粗多糖，纯化后测定羊栖菜多糖含量，并分析其单糖组分，为今后羊栖菜化学及药理研究提供有益参考。方法：水提醇沉法提取粗多糖，Sevage法除蛋白，H2O2脱色，流水透析。苯酚-硫酸显色法在490hm处测定其含量。高效毛细管电泳测定多糖组分。结果：羊栖菜中多糖含量达53．46％，平均加样回收率为99．37％。多糖由鼠李糖，木糖，葡萄糖，甘露糖，果糖，岩藻糖，半乳糖和一未知成分组成。结论：苯酚-硫酸法实验操作简便，准确，可用于羊栖菜多糖含量的测定。毛细管电泳分离度高，可用于多糖中单糖组分分析。     

Use of Sargassum fusiforme Extract and its Bioactive Molecules to inhibit HIV Infection: Bridging Two Paradigms between Eastern and Western Medicine

AIDS is a global pandemic that has seen the development of novel and effective treatments to improve the quality of life of those infected and reduction in the spread of the disease. While great advancements have been made in HIV/AIDS therapeutics, there is still no cure or viable vaccine in development. The high rate of HIV-1 mutation contributing to virus immune escape, combined with increase in sexual transmission and the significant clinical therapeutics side effects of the currently available treatments highlights the need for novel therapeutics with broad anti-viral activity against both CXCR4(X4) and CCR5(R5)-tropic viruses. In our search for novel modalities against HIV infection, we investigated several aqueous extracts from Traditional Chinese Medicine(TCM) collection and identified Sargassum fusiforme(S. fusiforme) as a potent inhibitor of HIV infection. Following the Western approach of drug discovery and development, we isolated several bioactive molecules from S. fusiforme and determined their mechanism of action. TCM and Western approaches to disease treatment and drug development have been shown to be complementary to each other. The former provides a rich medicinal history for natural compounds that can have clinical impact on a variety of illnesses, while the latter enables the application of chemical informatics with rational drug design approach coupled to mechanistic underpinnings of such therapeutics. This multistep paradigm is demonstrated in the example of S. fusiforme as a treatment to prevent HIV infection, as described in this review.     

羊栖菜提取物的降血糖作用研究

目的研究羊栖菜提取物对正常及四氧嘧啶所致糖尿病小鼠血糖的影响。方法用四氧嘧啶糖尿病小鼠为模型,以血糖、糖耐量为指标研究羊栖菜提取物的降血糖作用。结果羊栖菜提取物对正常小鼠无降血糖作用;预防性给予羊栖菜粗多糖(350,700 mg.kg-1)后,四氧嘧啶糖尿病小鼠血糖显著降低;羊栖菜粗多糖和醇提物(700 mg.kg-1)有治疗作用,能明显降低糖尿病小鼠血糖水平;对正常小鼠和糖尿病小鼠糖耐量无明显改善。结论羊栖菜提取物对四氧嘧啶糖尿病小鼠具有防治作用。     

半叶马尾藻多糖对小鼠S180肉瘤的抑制作用及其机制

背景与目的:由于目前使用的化疗药物在肿瘤治疗中副作用较大,严重制约了化疗药物在肿瘤治疗方面的应用。本实验目的是研究野生半叶马尾藻多糖(SP)对S180肉瘤小鼠肿瘤生长的抑制作用和机制,为寻找低毒和高效的治疗肿瘤药物提供有价值的实验资料。方法:称重法检测SP对小鼠S180肉瘤的抑瘤率,流式细胞术分析SP对S180肉瘤小鼠胸腺细胞和脾淋巴细胞周期的影响,3H-TdR和3H-TyR掺入法测定S180肉瘤小鼠胸腺细胞、脾淋巴细胞DNA合成及蛋白质合成,脾淋巴细胞转化实验检测SP对S180肉瘤小鼠的免疫增强作用。结果:在肿瘤接种前10天给予小鼠SP50、100和200mg/kg,抑瘤率分别为28.7%、32.9%和50.3%,其中100mg/kg和200mg/kg的SP组与NS对照组比较抑瘤率有显著性差异(P<0.05)。小鼠接种肿瘤细胞后,脾脏和胸腺明显缩小,其脏器指数与正常对照组比较有显著性差异(P<0.05)。SP可以缓解小鼠脾淋巴细胞和胸腺细胞在接种肿瘤细胞后发生的G1期阻滞。各浓度SP组小鼠胸腺细胞3H-TdR掺入值显著高于NS对照组。200mg/kgSP组小鼠脾细胞DNA合成及100mg/kg和200mg/kgSP组小鼠脾细胞蛋白质合成显著高于NS对照组,说明SP对肿瘤接种后引起的小鼠脾细胞DNA和蛋白质合成显著降低也有拮抗作用。各浓度SP组刀豆素A(concanavalinA,ConA)和脂多糖(lipopolysaccharide,LPS)诱导的小鼠脾淋巴细胞增殖显著高于NS对照组。结论:SP对S180肉瘤小鼠肿瘤生长有抑制作用,其机制可能是通过提高小鼠的脾淋巴细胞和胸腺细胞的数量和功能而实现的。     

两种海藻多糖的提取分析及其体外抗肿瘤作用

对采集于湛江硇洲岛的匍枝马尾藻(Sargassum polycystum C)和铜藻(Sargassum horneri)两种海藻进行多糖的热水提取，Sevag法除蛋白，通过紫外和红外光谱相互验证分析，证明其主要为多糖成分，并对其多糖成分进行体外抗肿瘤(人肺腺癌细胞系SPC-A-I)作用的初步研究，结果表明，终浓度为0．12％的匍枝马尾藻和铜藻的初筛抑瘤率分别为58．9％和32．9％。     

网格状与线状鼠尾藻多糖抗炎作用研究

目的 对鼠尾藻多糖进行分离纯化，并对其理化特性、微观结构和对炎症因子表达的影响进行研究。方法 采用热水提取法及DEAE-Sepharose CL-6B和Sephacryl S-300 HR柱层析分离纯化鼠尾藻多糖，利用高效空间排阻色谱法（HPSEC）、气相色谱法（GC）、红外光谱法（FTIR）和透射电子显微镜（TEM）研究理化特性及微观结构，实时荧光定量PCR法研究炎症相关因子的mRMA表达。结果 得到两种鼠尾藻均一多糖STP-I和STP-III，相对分子质量分别是29 和350 kDa，糖含量均超过96%。透射电子显微镜观察到STP-I是网格状结构，STP-III是线状且螺旋缠绕结构。活性研究表明，STP-III能更好地抑制脂多糖（LPS）诱导下小鼠巨噬细胞RAW264.7系炎症因子TNF-α、IL-6和COX-2的mRNA表达量，在150 μg?mL1质量浓度下，STP-III对炎症因子TNF-α、IL-6和COX-2的mRNA表达抑制率均超过95%。结论 鼠尾藻多糖具有抗炎症因子表达的作用，可以作为潜在的抗炎药物开发。线状缠绕的鼠尾藻多糖STP-III较网格状的STP-I能更好地抑制炎症相关因子的表达。     

褐藻叶托马尾藻中的甘油酯类成分

采用稻瘟霉模型生物活性追踪方法，应用多种层析手段，从褐藻叶托马尾藻（Sargassum carpophyllum)中分离得到5个甘油酯类成分，应用多种波谱技术和化学方法，分别确定为：1－O十六碳酰基－3－O－（13Z－十八碳烯酰基）甘油酯（I）1，2，3－O－三910Z，13Z－十八碳二烯酰）甘油酯（II），（2S）－1－O－十六碳酰基2－O－十四碳酰基－3－O－β－D－吡喃半乳糖基甘油（Ⅲ），（2S）－1－O－十六碳酰基－2－O－十六碳酰基－3－O－β－D－吡喃半乳糖基甘油（Ⅳ）和（2S）－1－O－十六碳酰基－2－O（11Z－十八碳烯酰基）－3－O－β－D－吡喃半乳糖基甘油（V），5个化合物均为首次从该各频繁 藻中得到，I和V为新天然产物，Ⅱ-Ⅴ具有诱导稻瘟霉菌丝变形活性和弱的肿瘤细胞毒性。     

Defensive nature of Sargassum polycystum (Brown alga)against acetaminophen-induced toxic hepatitis in rats: Role of drug metabolizing microsomal enzyme system, tumor necrosis factor-α and fate of liver cell structural integrity

AIM: To assess the defensive nature of Sargassum polycystum (S. Polycystum) (Brown alga) against acetaminophen (AAP)-induced changes in drug metabolizing microsomal enzyme system, tumor necrosis factor (TNF-α)and fine structural features of the liver during toxic hepatitis in rats.METHODS: Male albino Wistar strain rats used for the study were randomly categorized into 4 groups. Group Ⅰ consisted of normal control rats fed with standard diet.Group Ⅱ rats were administered with acetaminophen (800 mg/kg body weight, intraperitoneally). Group Ⅲ rats were pre-treated with S. Polycystum extract alone.Group Ⅳ rats were orally pre-treated with S. Polycystum extract (200 mg/kg body weight for 21 d) prior to acetaminophen induction (800 mg/kg body weight,intraperitoneally). Serum separated and liver was excised and microsomal fraction was isolated for assaying cytochrome P450, NADPH Cyt P450 reductase and b5.Serum TNF-α was detected using ELISA. Fine structural features of liver were examined by transmission electron microscopy.RESULTS: Rats intoxicated with acetaminophen showed considerable impairment in the activities of drug metabolizing microsomal enzymes, such as cytochrome P450, NADPH Cyt P450 reductase and b5 when compared with the control rats. The rats intoxicated with acetaminophen also significantly triggered serum TNF-α when compared with the control rats. These severe alterations in the drug metabolizing enzymes were appreciably prevented in the rats pretreated with S. Polycystum. The rats pretreated with S. Polycystum showed considerable inhibition in the elevation of TNF-α compared to the rats intoxicated with acetaminophen. The electron microscopic observation showed considerable loss of structural integrity of the endoplasmic reticulum, lipid infiltration and ballooning of mitochondria in the acetaminophen-intoxicated rats,whereas the rats treated with S. Polycystum showed considerable protection against acetaminophen-induced alterations in structural integrity.CONCLUSION: These observations suggest that the animals treated with S. Polycystum extract may have the ability to protect the drug metabolizing enzyme system and mitochondrial functional status from free radical attack, thereby showing its defense mechanism in protecting hepatic cells from acetaminophen toxic metabolite N-acetyl-para-benzoquinone-imine (NAPQI).