绵羊磷脂酶C-γ1对绵羊卵母细胞体外成熟的影响

卵母细胞成熟率可作为衡量体外培养卵母细胞质量和发育能力的指标。本研究旨在探讨PLC-γ1是否参与了绵羊卵母细胞的体外成熟及其影响。将绵羊卵母细胞在含有不同浓度的U73122（PLC抑制剂））及m-3M3FBS （PLC激活剂）的成熟培养液中进行培养,每个浓度150个细胞,重复试验3次,统计细胞成熟率、卵裂率及桑葚胚率以供筛选出PLC-γ1的最佳抑制及促进浓度。qPCR检测0.5 μmol·L^-1 U73122及0.5 μmol·L^-1 m-3M3FBS处理后48 h的卵母细胞中PLC-γ1、BAK、BAX、CASP3、CASP8、P53、BCL6基因mRNA水平表达情况,Western blot检测0.5 μmol·L^-1 U73122及0.5 μmol·L^-1 m-3M3FBS处理后48 h的卵母细胞中PLC-γ1、BAK、BAX、CASP3、CASP8、P53、BCL6蛋白的表达情况。每个浓度150个细胞,重复试验3次。在绵羊卵母细胞中,0.5 μmol·L^-1 U73122是促进成熟的最佳浓度,0.5 μmol·L^-1 m-3M3FBS是抑制成熟的最佳浓度。0.5 μmol·L^-1 U73122处理组卵裂率和囊胚率均显著低于对照组。0.5 μmol·L^-1 m-3M3FBS处理组卵裂率和囊胚率均显著高于对照组。通过qPCR检测结果显示,与对照组相比,U73122组中BAK、BAX、CASP3、CASP8、P53基因mRNA表达量显著上升,PLC-γ1、BCL6基因mRNA表达量显著下降;m-3M3FBS组则与之相反。Western blot结果显示,与对照组相比,U73122组中BAK、BAX、CASP8蛋白表达量显著增多,PLC-γ1、BCL6蛋白表达量显著减少,P53和CASP3蛋白表达量无明显变化。m-3M3FBS组中PLC-γ1、BCL6蛋白表达量显著增多,BAX、CASP3、P53蛋白表达量显著减少,BAK、CASP8蛋白表达量无明显变化。综上所述,本研究表明PLC-γ1在绵羊卵母细胞的体外成熟培养中发挥重要作用,其可调节卵母细胞的成熟以及早期胚胎的发育能力。     

Study on the hypoglycemic activities and metabolism of alcohol extract of Alismatis Rhizoma

The Alisma rhizoma is widely used in the therapy of diabetes in traditional folk medicine of China. Compositional analysis of the alcohol extract of Alismatis Rhizoma (AEA) revealed that the eight compounds gotten from AEA are all belonging to protostane-type triterpenes. The AEA and compounds were incubated with 3T3-L1 preadipocytes, glucose level in the 3T3-L1 adipocytes culture medium and lipid content in 3T3-L1 adipocytes were measured, and analysis of alpha-glucosidase inhibition of AEA and compounds. At the same time, the uptake of AEA by 3T3-L1 adipocytes and the metabolism of AEA in SD rats were analyzed by HPLC-ESI/MS. As result, AEA increased glucose uptake in 3T3-L1 adipocyte model, not increase adipogenesis; AEA inhibited alpha-glucosidase activity; alisol A-24-aceate (8) was absorbed by 3T3-L1 adipocytes; and two compounds were detected in blood and three were detected in urine in SD rats. So AEA had protostane-type triterpenes, these type compounds in AEA may have hypoglycemic activity via inhibition of alpha-glucosidase activity and promotion of glucose uptake. In contrast to the anti-diabetic drug thiazolidinediones, they did not induce adipogenesis, avoiding the displeased effects of rosiglitazone.     

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.     

海洋贝类提取物对血管平滑肌细胞增殖的影响

采用20％胎牛血清（FBS），碱性成纤维细胞生长因子（bFGF)所诱导的大鼠VSMCs增殖模型，观察了海洋贝类提取物（ESs)对血管平滑肌细胞（VSMCs)增殖的影响。将培养的5－8代细胞接种于96孔板，除正常组加入含0．4％FBS的M199培养液外，其他组均加入含20％FBS培养液（或含50ng/ml bFGF的0．4％FBS培养液），同时给药组分别加入含有25，50，100μl/ml浓度的ESs，孵育48h后，以细胞计数，结晶紫染色及MTT比色法VSMCc生长情况。结果表明，与模型组比较，ESs各剂量组VSMCs的增殖受到明显抑制（P＜0．05，P＜0．01）。ESs对20％FBS和bFGF所致的VSMCs增殖具有明显抑制作用，并存在剂量依赖关系。     

血清浓度对鸡胚盲肠上皮细胞原代培养的影响

为探讨细胞培养基中血清浓度对鸡盲肠上皮细胞体外培养的影响,分别用含0%、2.5%、5%和10%胎牛血清(FBS)的细胞培养液培养鸡胚盲肠上皮细胞,检测细胞活性;选择适宜的FBS浓度,并测定了该浓度条件下鸡盲肠上皮细胞的生长特性。结果表明:用含2.5%和5%FBS的培养基培养鸡胚盲肠上皮细胞,第4天细胞贴壁率达最高,分别为84%和80.33%,显著地高于无血清和10%FBS组(P<0.01/0.05),第7天仍均维持在76%以上,两者间除了第2天外,其余差异均不显著;用含2.5%FBS的细胞培养基进行鸡胚盲肠上皮细胞原代培养,第2~3天为适应阶段,第3~5天处于对数生长期,第5~9天细胞进入减速期,第10~11天再次进入对数生长期,第11天以后细胞进入衰退期,整个过程持续14 d左右。鸡胚盲肠上皮细胞培养的血清浓度以2.5%FBS为宜。     

Ethyl p-methoxycinnamate isolated from a traditional anti-tuberculosis medicinal herb inhibits drug resistant strains of Mycobacterium tuberculosis in vitro

Many plants are used in Ayurveda for the treatment of tuberculosis. Our aim was to examine if these plants possess any specific molecule that inhibits Mycobacterium tuberculosis. One of them, Kaempferia galanga, yielded an anti-TB molecule, ethyl p-methoxycinnamate (EPMC). By resazurin microtitre assay (REMA), EPMC was shown to inhibit M. tuberculosis H37Ra, H37Rv, drug susceptible and multidrug resistant (MDR) clinical isolates (MIC 0.242-0.485 mM). No cross resistance was observed to any standard anti-TB drugs in the MDR strains. The compound did not inhibit any prototype bacteria tested. EPMC seems to be a potential anti-TB lead molecule.     

胎牛血清及热处理家蚕体液对家蚕微孢子虫体外增殖的影响

对家蚕微孢子虫在体外增殖的研究表明 :培养基中胎牛血清 (FBS)添加量在 5 %～ 15 %范围内与微孢子产生量呈线性正相关关系 ;加热家蚕体液对家蚕微孢子虫体外增殖的促进作用则主要表现在与FBS的复合效应上 ,5 %FBS +5 %加热家蚕体液添加区的微孢子产生数为 10 %FBS添加区的 133 5 2 %。     

胎牛血清不同组分对昆虫杆状病毒复制的影响

为探讨胎牛血清(FBS)中的不同组分对昆虫杆状病毒(BAC)复制的影响,找到血清中能提高病毒复制效果的有效组分,对亲和凝胶层析柱法分离获得血清中的四个混合组分(S1、S2、S3和S4)进行了病毒培养实验。在6孔板上接种SF9细胞,培养24 h,接种0.01MOI的BAC-GFP病毒,按照0、0.3%、0.6%、1.3%、2.5%、5%的浓度梯度加入6孔板中进行病毒培养。分别在24 h、48h、72 h、96 h进行荧光拍照、观察。96 h收集病毒上清,进行空斑形成实验测定病毒滴度。用收集的病毒感染SF9细胞,72 h流式检测病毒的复制情况。结果显示:血清样本S1在浓度1.3%以上时,对于BAC的复制有促进作用; S2没有促进效果,反而随着S2的浓度上升而出现抑制的情况; S3在浓度2.5%以上时,同样有着明显促进作用; S4的促进作用不显著。试验表明,与原胎牛血清相比,添加单一血清组分S1对提高BAC病毒的复制效果更显著。     

EGF和不同血清对牛卵母细胞成熟及核移植胚体外发育的影响

分离牛卵母细胞,分别用添加表皮生长因子(EGF)和发情牛血清(发情当天血清(OCS(0 d))和发情第7天血清(OCS(7 d)))的培养液进行成熟培养后去核,构建牛体细胞核移植胚并进行体外培养,研究EGF和OCS对卵母细胞成熟及核移植胚体外发育的影响。结果表明EGF添加组和OCS(0 d)添加组卵细胞的成熟率及核移植胚的卵裂率和囊胚率分别为74.5%,84.6%,25.9%和83.9%,78.8%,31.3%。在牛卵母细胞成熟液中添加EGF、OCS(0 d)更有利于牛核移植胚胎的发育。     

小牛血清对耐辐射奇球菌类胡萝卜素在肿瘤细胞中吸收的影响

耐辐射奇球菌类胡萝卜素deinoxanthin的抗氧化活性高于β-类胡萝卜素等,并且是参与耐辐射奇球菌极端抗性的重要物质。但是,deinoxanthin溶液在体外对光、氧等因素不稳定。本文旨在研究deinoxanthin的有效溶剂载体系统,并以此载体研究deinoxanthin对人卵巢癌细胞系A2780的药理学作用。通...