Effect of selected aromatic medicinal plant sancho (Zanthoxylum schinifolium) on osteoblastic MC3T3-E1 cells

This study was performed to investigate the bioactivity of sancho (Zanthoxylum schinifolium) essential oil (EO) on bone metabolism and its function of osteoblastic MC3T3-E1 cells. The volatile aroma components of sancho EO were collected using a Clevenger-type apparatus by steam distillation extraction method, and determined by GC-MS. β-Phellandrene (22.54%) was the most abundant volatile compound in sancho EO, followed by citronellal (16.48%) and geranyl acetate (11.39%). It increased the collagen and mineralization of osteoblasts (p<0.05), indicating that sancho may help prevent osteoporosis.     

Effects of medicinal plant Atractylodes japonica on MC3T3-E1 cells

During our survey of the aromatic medicinal herbaceous plants looking for activity on bone metabolism, we found that Atractylodes japonica rhizomes increased the function of osteoblastic MC3T3-E1 cells. Chemical constituents of Atractylodes japonica were separated by hydro-distillation extraction (HDE), and characterized by GC/MS. The effects of Atractylodes japonica essential oils (AEOs) on the function of osteoblastic MC3T3-E1 cells were tested. AEO significantly (p<0.05) increased the growth and differentiation of osteoblastic MC3T3-E1 cells, indicating that has potential for use as a natural treatment for osteoporosis.     

Activities of MC3T3-E1 cells cultured on gamma-irradiated salmon atelocollagen scaffold

We investigated the effect of gamma-irradiation on the activities of MC3T3-E1 cells cultured on gamma-irradiated salmon atelocollagen (SAC) scaffolds. SAC-cultured samples were irradiated at doses of 10, 15, and 25 kGy. Gamma-irradiation had a significant impact on the activities of MC3T3-E1 cells. The proliferation rates and alkaline phosphatase activities of MC3T3-E1 cells increased with gamma-irradiation dose.     

Acteoside inhibits irradiation-mediated decreases in the viability and DNA synthesis of MC3T3-E1 cells

Therapeutic irradiation can cause bone loss, whereas antioxidant supplementation is considered to attenuate irradiation-mediated damages. This study examined whether or not acteoside inhibits irradiation-mediated changes in viability and proliferation of MC3T3-E1 cells. X-ray radiation at >4 Gy not only decreased cell viability and DNA synthesis in the cells, but also increased intracellular levels of reactive oxygen species (ROS) and phosphorylated p66^Shc protein. Irradiation at 8Gy also decreased intracellular levels of reduced glutathione (GSH) and induced G₁ phase arrest of cell cycle progression with the attendant increase of p21 induction. Pretreatment with acteoside inhibited the irradiation-mediated decreases in viability and DNA synthesis by restoring the radiation-mediated changes in the levels of ROS, GSH, p21, and p-p66^Shc to the untreated control levels. These inhibitory activities of acteoside were greater than that of a synthetic antioxidant compound or N-acetyl cysteine did. Collectively, acteoside treatment may prevent irradiation-induced oxidative damages to osteoblasts.     

卵黄高磷蛋白对壳聚糖膜基质上MC3T3-E1细胞增殖的影响

本研究制备了壳聚糖和氧化石墨烯/壳聚糖复合膜,并采用CCK-8法、Annexin V/PI双染法和PI染色检测了蛋黄中提取的卵黄高磷蛋白对MC3T3-E1细胞在该复合膜基质上的生理活动的影响。结果表明:壳聚糖膜对细胞无毒性影响,并通过提高S期细胞所占比例,使MC3T3-E1细胞活性提高了31.92%,抑制细胞凋亡,使凋亡率下降45.78%;氧化石墨烯/壳聚糖膜会提高细胞的凋亡率,氧化石墨烯含量越高,促进凋亡越明显,凋亡率在氧化石墨烯含量为2%时最高,为17.64%;在壳聚糖膜基质上,100 μg/mL卵黄高磷蛋白对MC3T3-E1细胞促进增殖作用最显著(p<0.05),细胞活性提高81.59%,同时降低了细胞的凋亡比例;蛋白浓度为500 μg/mL时,细胞活性下降4.47%,促进了细胞凋亡。综上所述,壳聚糖和低浓度卵黄高磷蛋白均能提高MC3T3-E1细胞的活性。     

金枪鱼鱼骨活性钙对鼠胚胎成骨细胞前体细胞(MC3T3-E1)活力和凋亡作用

目的:以金枪鱼骨为原料,制备柠檬酸-苹果酸钙剂(CMC),研究其对小鼠胚胎成骨细胞前体细胞(MC3T3-E1)的细胞活力、凋亡的影响,并探讨其作用机制。方法:在小鼠胚胎成骨细胞前体细胞(MC3T3-E1)培养液中,分别加入高浓度(1 μg/mL)CMC和低浓度(0.1 μg/mL)CMC,利用MTT法检测对MC3T3-E1细胞活力的影响,流式细胞仪检测血清饥饿诱导的细胞凋亡。结果:高浓度和低浓度的钙对MC3T3-E1细胞活力均有显著的促进作用,24 h的高钙组的细胞增长率(9.67%)优于低钙组(8.95%);36 h和48 h后低钙组的增长率分别为14.96%和20.33%,明显优于36 h和48 h作用后的高钙组(6.23%和1.73%),两组细胞凋亡率与空白组比较差异较小,且均显著低于对照组(p<0.01)。结论:CMC高钙组和CMC低钙组在一定浓度范围内,能促进MC3T3-E1细胞活力显著增强,并且改善血清饥饿诱导的成骨前体细胞凋亡。     

鸡蛋黄蛋白质水解产物制备及其对MC3T3-E1细胞增殖分化的影响

鸡蛋黄经脱脂得到蛋黄蛋白质,直接以MC3T3-E1细胞增殖活力为指标,对水解条件进行优化,从而制备蛋黄蛋白质水解产物(egg yolk proteins hydrolysate,EYPH)并对其促MC3T3-E1细胞增殖分化的活性进行研究。结果表明:在对比胰蛋白酶与风味蛋白酶水解过程中,发现单一胰蛋白酶反应得到产物活性最好,胰蛋白酶反应的最佳水解条件为水解时间6 h、水解温度37℃、酶与底物质量比1∶50、p H 8.0。EYPH使MC3T3-E1细胞增殖活力得到提高,为129.89%,细胞周期分析得到S期细胞增加了4.69%,这2个方面证实EYPH对MC3T3-E1细胞的增殖活力有显著的提高。EYPH对MC3T3-E1细胞分化矿化有显著影响,碱性磷酸酶活力提高了9.1%;刺激骨钙素分泌,其含量上升2 mg/mL;形成更多的矿化结节。此外,EYPH还能刺激RUNX2特异性转录因子的基因表达,表达量提高了6.12倍。结果说明EYPH对成骨细胞MC3T3-E1增殖分化有显著的促进作用,因此,EYPH对于抗骨质疏松方面有较好的应用潜力,为开发其功能性食品提供一定的数据参考。     

鲫鱼卵唾液酸糖蛋白化学结构分析及对前成骨细胞MC3T3-E1增殖分化的影响

目的：成熟的鲫鱼卵经提取、分离纯化得到唾液酸糖蛋白（sialyglycoproteins of Carassius auratus eggs,CA-SGP）,分析其化学结构,及对前成骨细胞MC3T3-E1增殖分化的影响。方法：采用水提法从成熟鲫鱼卵中提取水溶性蛋白,经阴离子交换色谱、凝胶过滤色谱分离纯化鲫鱼卵水溶性蛋白得到CA-SGP,测定其基本组成,采用高效凝胶液相色谱和聚丙烯酰胺凝胶电泳测定其分子质量和纯度,PMP柱前衍生高效液相色谱法测定其单糖组成;采用噻唑蓝法测定MC3T3-E1细胞的增殖率,酶联免疫法测定Ⅰ型胶原蛋白（collagen type Ⅰ,COLⅠ）和骨钙素（osteocalcin,OCN）的分泌量及碱性磷酸酶（alkaline phosphatase,ALP）的活性,茜素红染色法测定矿化结节的数量。结果：得到单一的组分CA-SGP,在高效凝胶过滤色谱色谱图上呈现单一的糖和蛋白的检测重叠峰,在凝胶电泳图上呈现单一弥散性条带,其蛋白质含量为14.33%,己糖含量为62.81%,N-乙酰神经氨酸（N-acetylneurainic acid,Neu5Ac）含量为19.72%。单糖组成测定结果显示：CA-SGP主要由甘露糖、葡萄糖胺、半乳糖胺组成,其物质的量比为7.61∶6.70∶1.00;MC3T3-E1细胞与CA-SGP共同孵育后,其增殖率显著增加,COLⅠ和OCN的分泌量分别增加了39.28%和83.06%,ALP活性提高了285.08%,矿化结节的数量增加了96.41%,说明CA-SGP显著促进MC3T3-E1细胞增殖、分化和矿化。结论：CA-SGP为富含Neu5Ac的糖蛋白,具有显著促进MC3T3-E1细胞增殖、分化和矿化的作用,对开发抗骨质疏松功能性食品具有重要意义。     

Effect of far infrared drying on antioxidant property, anti-inflammatory activity, and inhibitory activity in A549 cells of Gamguk (Chrysanthemum indicum L.) flower

This research demonstrated and compared the effects of shade and far infrared drying (FIRD) of gamguk (Chrysanthemum indicum L., CE) flowers extract on total phenolic (TP), total flavonoid (TF) content, anticancer and anti-inflammatory activities. The research data revealed that the TP and TF contents were highest in FIRD treated CE flower extracts. Similarly, the effect of CE on lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW264.7 cells showed in a dose dependent manner. Further, the CE inhibited the tumor necorsis factor (TNF)-α, cyclooxygenase (COX)-2 expression, and prostaglandis E₂ (PGE₂) production. The anticancer activity was monitored in A549 lung cancer cell, which showed that FIRD treated CE inhibited cell proliferation significantly (p<0.05) higher in dose and time dependent manner.     

牛骨肽钙螯合物制备、表征及其促MC3T3-E1细胞成骨活性

目的：为制备人体高效吸收利用的补钙剂并探究其对MC3T3-E1细胞成骨活性的影响。方法：对牛骨多肽进行钙螯合处理,以钙螯合能力为考察指标,通过响应面方法确定最佳制备条件;利用红外光谱、X射线衍射和原子吸收等方法探究肽钙螯合物的结构特性及其稳定性;通过细胞实验探究牛骨肽钙螯合物对MC3T3-E1细胞增殖、分化和矿化活性的影响。结果：制备牛骨肽钙螯合物的最佳条件为肽钙质量比2.97∶1、温度62.54℃、pH 9.06。在该条件下,钙螯合能力达到55.65 μg/mg。光谱结构分析结果表明氨基氮、羟基氧和羧基氧是钙主要螯合位点,二者之间主要通过配位键结合,且肽螯合钙后分子质量增加;此外,牛骨肽钙螯合物稳定性受温度影响较小,但对pH值较敏感。MC3T3-E1细胞实验结果表明肽钙螯合物具有明显促MC3T3-E1细胞增殖、分化和矿化活性的作用。结论：制备的牛骨肽钙螯合物具有潜在的促MC3T3-E1细胞成骨活性功效。     

郑州刺槐花挥发油化学成分分析

采用同时蒸馏萃取法提取郑州地区刺槐花挥发油,用GC-MS-DS联用技术分析其中化学成分,并与文献报道的其他地区刺槐花挥发油主要成分进行了对比.结果表明,郑州地区刺槐花挥发油中分离出64种组分,鉴定出48种化合物,占出峰总面积的82.93%;郑州地区刺槐花挥发油主要成分是6,10,14-三甲基-2-十五烷酮、3,7,11-三甲基-2,6,10-十二碳三烯-1-醇、2-氨基苯甲酸甲酯等,与文献报道的其他地区刺槐花主要成分和含量均有所不同.     

Effect of bovine lactoferrin and human lactoferrin on the proliferative activity of the osteoblast cell line MC3T3-E1 in vitro

We conducted a comparative in vitro study on the proliferative effects of natural human lactoferrin (nhLF) and bovine lactoferrin (bLF) on osteoblasts. We investigated cell proliferation, cell survival, cell cycle, and mRNA and protein expression of proliferating cell nuclear antigen. Results indicated that treatment with 100 μg/mL of bLF or nhLF promoted the proliferation and sustenance of osteoblasts, and increased the length of the G₂/M and S phases compared with the untreated osteoblasts. Results of real-time quantitative PCR and Western blot showed that mRNA and protein expression of proliferating cell nuclear antigen by osteoblasts treated with bLF or nhLF were greater than those of the untreated control. At the same concentration, bLF demonstrated a greater effect on osteoblast proliferation than did nhLF. This study provides insights of significance in the utlization of bLF in healthy food formulas.     

茼蒿精油提取工艺的研究

以茼蒿为原料,对茼蒿精油的提取方法进行了研究。通过不同溶剂提取效率的比较,确定乙醇为最佳提取溶剂,并对乙醇提取茼蒿精油的溶剂浓度、温度、时间和料液比进行单因素实验与正交设计实验,确定最佳提取工艺条件为:乙醇浓度85%,提取温度45℃,提取时间6.5h,料液比1∶15。     

大孔树脂对野菊花总黄酮吸附分离特性的影响

选择12 种大孔树脂,比较其对野菊花总黄酮的吸附和解吸性能。在此基础上,筛选出较优的3 种大孔树脂,通过研究其静态吸附动力学特性,进一步筛选出适合分离野菊花总黄酮的理想树脂,并考察pH 值和洗脱剂体积分数对其吸附容量和解吸率的影响。结果表明：XDA-1、LSA-21 和AB-8 树脂有较大的吸附容量和较高的解吸率,且其中XDA-1 树脂显著优于另外两种,其吸附容量高达84.1mg/g、解吸率为96.2%,而且具有良好的静态吸附动力学特性,是更为适合吸附分离野菊花总黄酮的理想树脂。XDA-1 树脂吸附分离野菊花总黄酮较合适的上样液pH 值为4～5,洗脱剂为70% 乙醇溶液。     

响应面法优化龟甲蛋白提取工艺及其对MC3T3-E1细胞增殖活性

目的：优化龟甲蛋白提取工艺,筛选龟甲促成骨细胞（MC3T3-E1）增殖活性组分。方法：以蛋白含量及对MC3T3-E1细胞增殖率为指标筛选龟甲蛋白提取方法;以料液比、提取温度、时间为自变量,龟甲蛋白含量为因变量,进行单因素提取研究,结合响应面试验设计,获得提取龟甲蛋白最佳工艺;通过超滤技术将龟甲蛋白按分子量分为5个不同组分：总提组分（组分1）,Mw>10 kDa组分（组分2）,Mw：3~10 kDa组分（组分3）,Mw：1~3 kDa组分（组分4）,Mw<1 kDa组分（组分5）,CCK8法测定不同浓度龟甲蛋白（12.5、25、50、100、200 μg/mL）及其不同组分对MC3T3-E1细胞增殖率的影响。结果：磁力搅拌方法得到的龟甲蛋白含量最高,且活性最佳,龟甲蛋白最佳提取条件为料液比1：13（g/mL）、提取温度30℃、提取3 h,此条件下测得龟甲蛋白含量为15.16%;龟甲蛋白不同组分均能促进MC3T3-E1前成骨样细胞增殖,并呈浓度依赖性,在浓度为200 μg/mL时,组分1~5对细胞的增殖率分别为20.58%、25.30%、30.24%、37.89%、38.15%,分子量小于1 kDa的组分促MC3T3-E1细胞增殖活性最佳。结论：本研究为龟甲蛋白的制备工艺提供参考,并证明龟甲蛋白及其不同组分均具有较好的促成骨细胞增殖活性,其中分子量小于1 kDa组分对MC3T3-E1细胞增殖的效果最佳。