In vitro antioxidant and wound healing activity of Sargassum polycystum hydroethanolic extract in fibroblasts and keratinocytes

Objective:To investigate the in vitro antioxidant and wound healing properties of the hydroethanolic extract of Sargassum polycystum,and elucidate the mechanism of its wound healing activity.Methods:Human dermal fibroblast and HaCaT cells were used to evaluate the proliferation by sulforhodamine B and dsDNA assay after treatment with Sargassum polycystum extracts.Scratch wound healing and phalloidin-rhodamine staining were employed to observe migratory activity and filopodia formation,respectively.Western blot and real-time RT-PCR assays were performed to determine the protein and gene expressions related to wound healing activities.Results:The phytochemical analysis found a higher level of flavonoid than phenolic compound in Sargassum polycystum extracts.In human dermal fibroblast cells,Sargassum polycystum extracts at 50 and 100μg/mL significantly increased fibroblast proliferation and the gene expressions of hyaluronic acid synthase 1(HAS1),HAS2,HAS3,collagen type 1 alpha 1 chain(COL1A1),collagen type 3 alpha 1 chain(COL3A1),and elastin.The phosphorylation of Akt,ERK1/2,and p38 MAPK was also significantly upregulated after treatment with Sargassum polycystum extracts.Additionally,50 and 100μg/mL of the extracts prominently enhanced the proliferation,migration,and filopodia formation of HaCaT cells,as well as the protein levels of pFAK/FAK,pSrc/Src,pAkt/Akt,pERK1/2/ERK1/2,Rac1 and Cdc42.Conclusions:Sargassum polycystum extracts show promising wound healing activities in human dermal fibroblasts and keratinocytes.

Defensive nature of Sargassum polycystum (Brown alga) against acetaminophen-induced toxic hepatitis in rats: Role of drug metabolizing microsomal enzyme system, tumor necrosis factor-a 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 bs. 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 bs when compared with the control rats. The rats intoxicated with acetaminophen also significantly triggered serum TNF-a when compared with the control rats. These severe alterations in the drug metabolizing enzymes were appreciably pretreated with S. polycystum. prevented in the rats The rats pretreated withS. 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）.

匍枝马尾藻中砷的脱除工艺研究

研究以匍枝马尾藻为原材料,借助电感耦合等离子体质谱技术(ICP-MS)分析了其中总砷含量,并评估了藻体粒径大小、脱除剂、料液比、脱除温度和时间等因素对总砷脱除效果的影响。在此基础上,通过正交优化试验,得到脱砷的最佳工艺参数为:藻粉粒径0.1 cm,脱除剂为水,料液比1∶40(g/mL),脱除时间3 h,脱除温度45℃,匍枝马尾藻中总砷的脱除率为85.42%。此工艺简单易操作,能够脱除马尾藻中大部分砷,为降低海藻中有害元素砷提供了有效途径。

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 polycystumC)和铜藻 (Sargassumhorneri )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 5 8.9%和 3 2 .9%。

匐枝马尾藻挥发油化学成分的研究

首次采用超临界CO2流体萃取技术(SFE-CO2 Method)萃取匐枝马尾藻挥发油,并用毛细管气相色谱-质谱(CGC-MS)联用技术对萃取的匐枝马尾藻挥发油的化学成分进行了研究。经毛细管气相色谱分离出27个峰,共确认了其中21种成分,占挥发油总量的95.36%。其主要成分是十六烷酸、7-Z-十六碳烯酸、9-E-十八碳烯酸、肉豆蔻酸、5,8,11,14-二十碳四烯酸乙酯、角鲨烯、邻苯二甲酸二辛酯等,为进一步开发利用匐枝马尾藻提供了科学依据。

匍枝马尾藻人工栽培初步研究

为探索匍枝马尾藻(Sargassum polycystum)的人工栽培技术,采用浮筏式栽培方式于2015年3—6月在海南省儋州市海头镇进行了匍枝马尾藻的人工栽培试验,记录了该藻的生长规律,并通过栽培水深和夹苗密度试验,比较了不同栽培条件对其生长的影响。结果显示,3月下旬至5月下旬为匍枝马尾藻生长最旺盛的季节,其中藻体长度的快速增长主要集中在4月中旬以前,而藻体重量以相对稳定的速率持续增长至5月下旬。栽培水深和夹苗密度分别对匍枝马尾藻的藻体长度和分枝数有显著影响,而藻体鲜重则同时受到这两个因素的影响(P<0.05)。研究表明,适量增加匍枝马尾藻的栽培水深和控制夹苗密度有利于促进其生长;为有效提高匍枝马尾藻的产量,栽培水深应设计为80 cm左右,夹苗密度控制在10 cm左右。

匍枝马尾藻有性繁殖和人工育苗的初步研究

为研究匍枝马尾藻有性繁育过程的特点和人工育苗的可行性,通过野外观察和室内培养观察的方法,对匍枝马尾藻雌雄生殖托特点、受精卵分化等有性繁育过程、以及假根的形成、幼苗发育进行了研究,此外还对匍枝马尾藻的人工育苗进行了初步探索。试验结果表明,匍枝马尾藻的繁殖季节集中在每年的5—6月份;水温达到28℃时,生殖托出现排精排卵现象;从受精卵发育到形成具有假根的幼苗约需要48h;幼苗发育缓慢,35d后出现原生叶。人工育苗结果显示,幼苗栽培35d,幼苗株高为2.5～3mm,密度为6～10株/cm2;海区栽培60d后,匍枝马尾藻幼苗株高可达到7cm,幼苗密度为2株/cm2。本试验结果为匍枝马尾藻的人工规模环境育苗提供了技术指导。