巴西苏木红素对血管平滑肌舒张作用的研究

巴西苏木红素为中药苏木的主要活性成分之一。利用体外血管舒缩试验以及血管平滑肌细胞试验,从分子生物学角度对巴西苏木红素舒血管效应及其机制进行了研究。结果表明,巴西苏木红素主要通过影响K+通道,抑制膜去极化,进而使血管舒张。这种效应与阻断5-HT受体有关。同时还与调控胞内Ca2+浓度,影响钙调蛋白,下调MLCP有关。这一过程中cAMP、PKA和PGI2也受巴西苏木红素影响并参与了调控。

HepG2^(IgG)转基因细胞应用于中药致敏物质预测研究

本文对HepG2IgG细胞对于Ⅱ型致敏性预测进行了较为深入系统的研究,其中包括转染宿主细胞的筛选、转染最佳条件、最适检测仪器以及转染持续时间等。并对己知致敏原及其市售的几种中药注射剂进行了致敏性筛选。结果表明,该细胞系具有检出灵敏性高,细胞转染成功率高,致敏因子表达稳定等特点。为早期预测II型致敏反应的中药提供了一个有价值的工具。

石榴叶鞣质与菠萝叶酚对哺乳期小鼠生长的比较

目的:观察石榴叶鞣质和菠萝叶酚通过母乳对哺乳期小鼠生长的影响。方法:给母鼠灌胃不同剂量的药物,观察新生小鼠的一般状况、体质量、尾长及脏器系数肝脏相关脂代谢调控因子的表达。结果:石榴叶鞣质和菠萝叶酚均可以通过母乳明显影响新生小鼠的生长发育,其中石榴叶鞣质的作用强于菠萝叶酚,同时可以明显下调新生小鼠肝脏的脂代谢相关酶的表达。结论:石榴叶鞣质和菠萝叶酚可以通过母乳影响新生小鼠的生长发育,其作用与影响肝脏脂代谢酶表达有关。哺乳期服用该药需引起注意。其临床意义尚有待于深入研究。

合浦珠母贝外套膜细胞engrailed、BMP2和Smad3的mRNA表达水平的相关性(英文)

软体动物engrailed蛋白和骨形成相关蛋白对胚胎贝壳区域边界形成可能具有重要作用,engrailed还被推测为调节基质蛋白在外套膜组织区域化表达的重要调控因子.因此,弄清调控engrailed在软体动物中特征表达的分子机制有着重要的研究意义.但是,由于贝类基因组测序尚不完整,目前也没有建立获得贝类细胞系,以致于许多预测可能参与调控的基因需要通过克隆来鉴定,而且经典的研究细胞信号通路的方法也很难得到应用.目前,在中国南海广泛养殖的合浦珠母贝中,已获知其BMP2和Smad3的cDNA全长,以该贝的基因组为模板,PCR扩增获得了一段engrailed编码区片段.经软件分析,该片段含有EH4结构域,且与其他物种engrailed蛋白具有很高的同源性.研究的贝中,特别是外套膜组织中,engrailed、BMP2和Smad3三者表达之间的相关性,将有助于我们理解贝壳形成的分子机制.贝壳缺刻后半定量PCR试验结果表明,三者均参与贝壳修复,且在贝壳缺刻后的修复过程中,engrailed和Smad3的mRNA表达变化规律非常相似,提示它们之间可能存在相互影响的联系.用地塞米松(DXM)和过氧化氢(H2O2)分别处理原代培养的贝外套膜组织迁出细胞,实时相对定量PCR检测engrailed、BMP2和Smad3的mRNA表达水平,统计分析结果表明,三者具有显著的相关性.上述所有结果为进一步研究贝类生物矿化的发育和信号转导机制提供了新的思路和基础.

大鼠N型钙通道ⅢP区原核表达、纯化及活性研究(英文)

高保真PCR克隆出编码大鼠海马组织N型钙通道α1B亚基ⅢP区的DNA序列,分别与表达载体pET28b和pGEX-4T-1连接,转化入能补充稀有密码子tRNAs的大肠杆菌Rosetta菌株中,通过优化表达条件,实现了高效诱导表达.再通过低温诱导,将包涵体用尿素溶解再稀释后透析法复性、酶切、亲和柱层析等方法得到较纯的可溶性目的蛋白HIS-Cav22P、GST-Cav22P和Cav22P,紫外吸收光谱检测实验证明了重组蛋白HIS-Cav22P能和钙离子可逆地结合.最后通过GST沉降实验证明了芋螺毒素SO3与Cav22P存在体外相互结合作用.上述结果为揭示芋螺毒素SO3特异性阻断大鼠海马组织N型钙流的分子机制提供了直接的依据,也为建立筛选新的非吗啡型天然镇痛药物的技术平台提供了基础.

甘遂致小鼠稀便与炎性分泌相关(英文)

目的:甘遂在我国有着几千年的应用历史,主要用于促进肠运动。通过实验观察甘遂通便促进肠推进同时是否伴有病理变化。方法:应用30%甘遂提取物给小鼠灌胃后排便及其大小肠的影响,进行组织学检查。同时观察大小肠IL-1β,TNFα和Caspase3的mRNA和蛋白表达的变化。结果:甘遂能够明显促进小鼠排稀便,抑制肠平滑肌收缩,肠粘膜出现明显炎性损伤,并伴有IL-1β和TNFα表达的上调。结论:甘遂促进排便与刺激肠粘膜分泌和炎性反应有关。

Ginsenoside Rg_1 reduced the invasion of S.aureus into respiratory epithelial cells involving pro-inflammatory cytokines and glucocorticoid receptor

The invasion of Staphylococcus aureus into respiratory epithelial cells and the followed inflammatory responses cause serious tissue damage.The aim of this study was to investigate the effects of ginsenoside Rg_1（Rg_1）on S.aureus infection in vitro and its action mechanism.An internalization model was constructed to determine the effect of Rg_1 on S.aureus invasion. The changes of expression of integrinβ1,NF-κB and glucocorticoid receptor were analyzed by Western blot.Expression of pro-inflammatory genes was validated using RT-PCR.The results demonstrated that Rg_1 treatment could reduce the invasion of S.aureus into rat pulmonary epithelial cells by down-regulating integrinβ1.Its anti-inflammatory action was exerted through reducing NF-κB and expressions of intercellular adhesion molecule-1（ICAM-1）,tumor necrosis factor-α（TNF-α）,interleukin-2 （IL-2）and interleukin-6（IL-6）.The increased expression of glucocorticoid receptor was involved in this regulation.The results suggested that Rg_1 could play a positive role in reducing S.aureus infections.Rg_1 could be used for the treatment of S.aureus infection,potentially.

Comprehensive study of rutaecarpine on vascular constriction relative to RhoA/MLCP-MLC signaling

The aims of the present study are to investigate the effect of vasoconstriction and to explore the mechanism of rutae- carpine. The research findings showed that rutaecarpine could induce contractions of the rat thoracic aorta in vitro. The inhibitors of Rho-kinase and inositol 1,4,5-triphosphate receptor （IP 3 R） could suppress the effect of rutaecarpine-induced vasoconstriction. In the study of A7r5 cells （a line of smooth muscle cells）, 300 μg/L rutaecarpine promoted the concentration of intracellular Ca 2＋ and enhanced the IP 3 R expression, which connects with 1,4,5-triphosphate to evoke the release of Ca 2＋ from the intracellular stores. Rutaecarpine increased the RhoA mRNA expression when the cells were pretreated with inhibitor H-1152, and improved the levels of phosphorylation of myosin light chain phosphatase （MLCP） and myosin light chain （MLC）. These results suggest that rutaecarpine plays a role in vasoconstriction relative to the RhoA/MLCP-MLC signaling pathway, which denotes a new field of rutaecarpine in pharmacology.

Comparison of berberine and its five analogues on cell viability and COX-2expression during glucose-oxygen deprivation and reperfusion in PC12 cells

Berberine, an isoquinoline alkaloid component of Rhizoma Coptidis has been demonstrated to be the key active ingredient involved in its protective effect against cerebral ischemia-reperfusion. However, the comparison among the analogues to the protective effect against oxygen and glucose deprivation/reoxygenation （OGD-R） was mediated by inhibition of cyclooxygenase-2 （COX-2） has never been reported. The aim of this study is to investigate the protective effect of berberine and its five analogues against OGD-R in PC 12 cells, as well as to determine whether the protective effect was regulated through COX-2. An established in vitro OGD-R model of PC12 cells by oxygen glucose deprivation of 4 h and reperfusion of 24 h was used in our study. After cells were treated with berberine or its five analogues, we examined the cell viability assay by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） assay. Cells were also collected to determine the levels of mRNA and protein of COX-2 by real time PCR and Western blot. We found that berberine and its analogues improved the viability of PC12 cells against OGD-R. Whereas berberine and berberrubine presented stronger activity with the most effective dose of 0.31 lag/mL and the minimum effective doses of 0.02 and 0.04 gg/mL. Palmatine possessed potentially weaker protective effect. The mRNA level of COX-2 in cells treated with berberine, coptisine and epiberberine was decreased significantly. The protein level of COX-2 was significantly down-regulated in cells treated with berberine. Studies suggested the important role of methylenedioxy groups （R2 and R3） of berberine analogues in COX-2 inhibitory effect, and methylenedioxy groups （R2, R3, R9 and R10） in berberine analogues in binding affinity with COX-2. Substituted hydroxyl group at R9 did not affect the activity of berberine. In summary, our study illustrated the protective effects of berberine and its analogues in PCI2 cells against OGD-R and to elucidate the structure-activity relationships. Docking analysis indicates that methylenedioxys at R2 and R3 is involved in the effect. More studies in other cells are needed to confirm our results.