Simplified tetrandrine congeners as possible antihypertensive agents with a dual mechanism of action

A series of O- and/or N-substituted derivatives of (+/-)-coclaurine (1a) were synthesized as simplified structural mimics of the antihypertensive alkaloid tetrandrine (2) and assayed for binding to brain cortical sites labeled with the alpha(1)-adrenergic radioligand [(3)H]prazosin or the calcium channel radioligand [(3)H]diltiazem. The introduction of O-benzyl groups on the coclaurine molecule, which exhibits only adrenergic antagonist activity, led to the appearance of calcium channel blocking activity comparable to that of tetrandrine while retaining adrenolytic activity in the same concentration range. Contraction of aortal rings with noradrenaline or KCl was relaxed more potently by some of these coclaurine derivatives than by tetrandrine, suggesting leads for the development of novel antihypertensive drugs with a dual mechanism of action.     

三氧化二砷对MRL/lpr小鼠CD4+T细胞增殖和IFN-γ、IL-4分泌的调节作用

 目的探讨三氧化二砷(ATO)对MRL/lpr狼疮小鼠脾脏CD4^+T细胞增殖以及IFN-γ和IL-4转录和翻译水平的影响。方法免疫磁珠分别分选15只MRL/lpr狼疮小鼠和15只C57BL正常对照小鼠脾脏CD4⁺T细胞,进而将CD4⁺T细胞随机分为空白组(RPMI1640),对照组(PHA20mg·L^-1+IL-21000u·mL^-1),ATO组[PHA20mg·L^-1+IL-21000u·mL^-1(48h)后+ATO1.0μmol·L^-1],体外共培养至72h。采用细胞计数试剂盒(CCK-8)检测3组CD4⁺T细胞的增殖和抑制水平;酶联免疫吸附反应(ELISA)检测上清液中IFN-γ和IL-4水平;逆转录-聚合酶链反应(RT-PCR)检测上述细胞因子基因的转录水平。结果①分选后获得的CD4⁺T细胞纯度为(94.12±1.08)%,细胞存活率为(95.82±3.41)%;②MRL/lpr小鼠对照组CD4⁺T细胞的增殖水平(A值)(0.343±0.019)较空白组(0.054±0.008)高(P<0.001);却低于C57BL小鼠对照组的水平(0.375±0.018)(P<0.05);③MRL/lpr和C57BL小鼠ATO组CD4⁺T细胞增殖水平(A值)分别为(0.324±0.013)和(0.362±0.013),均低于对照组(P<0.05,P<0.05);④MRL/lpr小鼠对照组IFN-γ的表达水平[(562.287±60.345)ng·L^-1]明显高于C57BL小鼠对照组[(253.208±59.434)ng·L^-1](P<0.05);ATO对MRL/lpr小鼠IFN-γ的抑制程度大于C57BL小鼠(P<0.05);MRL/lpr小鼠对照组的IL-4表达水平[(41.332±8.039)ng·L^-1]与ATO组[(23.046±3.253)ng·L^-1]和C57BL小鼠对照组[(29.303±8.038)ng·L^-1]比较,也具有明显的统计学差异(P<0.05和P<0.05);ATO对MRL/lpr小鼠IL-4的抑制程度高于C57BL小鼠(P<0.05);⑤在转录水平上,与C57BL小鼠相比,ATO对MRL/lpr小鼠IFN-γmRNA和IL-4mRNA的抑制程度更加明显(P<0.05和P<0.05);ATO对IFN-γ和IL-4表达水平的抑制和转录水平的抑制正相关,r=0.720(P<0.001)和r=0.557(P<0.01)。结论MRL/lpr小鼠存在CD4⁺T细胞的异常增殖,ATO能在不影响细胞存活率的浓度下抑制MRL/lpr小鼠CD4⁺T细胞的异常增殖活化,通过抑制其细胞因子IFN-γ和IL-4的mRNA分泌影响两者的表达,且对MRL/lpr小鼠抑制程度大于C57BL小鼠,表明ATO能抑制狼疮鼠的异常免疫功能,可进一步用于SLE的治疗。     

Rubia cordifolia extract inhibits cell proliferation in A-431 cells

The antiproliferative property of Rubia cordifolia (Rubiaceae) extract has been studied on two different cell types, A-431 cells (epidermal carcinomoid cells) and 3T3 fibroblast cells. It was observed that a fraction of Rubia cordifolia significantly inhibited the incorporation of [³H]-thymidine, induced by fetal bovine serum, in a dose-dependent manner. It also inhibited the PMA (phorbol 12-myristate 13-acetate) induced expression of c-fos genes in A-431 cells. It appears that inhibition of DNA synthesis underlies the mechanism for its antiproliferative properties. © 1998 John Wiley & Sons, Ltd.     

益肾养阴合剂通过CXCR4/STAT3信号通路对MRL/lpr小鼠肾脏发挥保护作用

目的:研究益肾养阴合剂对系统性红斑狼疮小鼠肾脏的保护作用并探讨其作用机制。方法:MRL/lpr疾病模式小鼠饲养发病后,益肾养阴合剂灌胃给药(17.25 g·kg-1),同时阳性药组用醋酸泼尼松灌胃给药(0.65 mg·kg-1),正常C57背景小鼠和实验MRL/lpr小鼠组灌胃同体积生理盐水,每天2次,连续给药28 d,每7 d收集尿液,检测尿蛋白变化情况;第29天取材肾脏组织,免疫荧光染色检测免疫球蛋白G(IgG)沉积情况;Raybiotech抗体芯片检测308个细胞因子变化情况,酶联免疫吸附测定(ELISA)验证部分变化因子(小鼠肾脏组织与患者血清水平);蛋白免疫印迹法(Western blot)检测Janus激酶2/信号转导和转录活化蛋白3(JAK2/STAT3)信号的磷酸化水平、基质细胞衍生因子1/趋化因子受体4(SDF1/CXCR4)信号轴、辅助性T细胞17(Th17)分泌因子白细胞介素-17(IL-17)的表达情况。结果:与MRL/lpr组比较,在中药和激素给药组,随着给药时间延长,尿蛋白含量逐渐降低,在第4周变化最为明显(P0.05),在给药28 d后,益肾养阴合剂组与醋酸泼尼松组的肾脏IgG沉积均有减轻(P0.05)。经Raybiotech抗体芯片筛查后发现,与C57正常小鼠,MRL/lpr疾病小鼠的肾脏SDF1,CXCR4蛋白信号表达有显著增强(P0.01);与MRL/lpr疾病小鼠比较,益肾养阴合剂组的小鼠肾脏SDF1,CXCR4蛋白信号表达显著降低(P0.01),经大样本量ELISA实验验证后,SDF1/CXCR4蛋白的抗体芯片结果可靠,进一步经Western blot实验检测发现,与正常C57小鼠比较,MRL/lpr小鼠肾脏组织中JAK2/STAT3信号通路被激活,其磷酸化蛋白表达增高(P0.05),SDF1,CXCR4,IL-17蛋白表达明显升高(P0.05);与模型小鼠比较,给予中药和激素灌胃处理28 d后JAK2,STAT3的磷酸化水平明显降低(P0.05),SDF1,CXCR4蛋白表达下降(P0.05),IL-17的表达也有明显的减少(P0.05)。结论:益肾养阴合剂可降低MRL/lpr模型小鼠的尿蛋白水平,其可通过抑制JAK2/STAT3与SDF1/CXCR4信号通路的激活,降低辅助性T细胞17的活性,减少IL-17的分泌,起到肾脏保护作用。     

高效液相-串联质谱测定血浆中依西美坦的浓度

 目的建立高效液相色谱-串联质谱的方法测定人血浆内的依西美坦浓度,对两种制剂进行人体生物等效性研究。方法采用Lichrospher C₁₈(4.6mm×150mm,5μm)色谱柱;柱温25℃;流动相为甲醇-水(含0.05%甲酸)(80∶20);流速为1.0mL·min^-1;通过液相串联质谱,大气压化学电离(APCI),以选择反应监测(SRM)方式进行检测;检测离子为依西美坦m/z 296.8[M+H]⁺→121[M+H]⁺,非那雄胺(内标)m/z373.2[M+H]⁺→305.2[M+H]⁺。结果依西美坦的线性范围为0.0994～39.76μg·L^-1,依西美坦受试制剂和参比制剂的t_1/2分别为(13.37±4.62)和(15.65±5.89)h,ρ^max分别为(19.97±8.51)和(20.04±9.16)μg·L^-1,t_max分别为(0.95±0.39)和(0.90±0.29)h,AUC_0-72分别为(95.64±26.87)和(96.73±25.97)μg·h·L^-1,AUC_0-∞分别为(100.47±28.76)和(114.59±62.32)μg·h·L^-1。结论该方法简便、灵敏度高,两制剂生物等效。     

拟黑多刺蚁活性组分对系统性红斑狼疮的治疗作用及机制

目的:采用动物模型和细胞模型,探讨拟黑多刺蚁活性组分(AFPV)对系统性红斑狼疮(SLE)的作用及可能机制。方法:采用大鼠Arthus反应,将60只SD大鼠随机分为正常组,模型组,醋酸泼尼松组(5 mg·kg~(-1)),AFPV高、中、低剂量组(400,200,100 mg·kg~(-1))。采用牛血清蛋白-弗氏完全(不完全)佐剂复制SLE模型。观察AFPV对SLE大鼠背部皮肤红肿直径的影响,酶联免疫吸附测定检测血清抗双链DNA(ds DNA)抗体,补体3(C3),补体4(C4),免疫球蛋白M(Ig M),白细胞介素-1(IL-1),白细胞介素-6(IL-6),白细胞介素-31(IL-31),白细胞介素-33(IL-33)水平;免疫磁珠法分选16～18周MRL/lpr狼疮小鼠和C57BL/6J正常小鼠脾脏CD4+T细胞,观察AFPV对miR-200a,miR-155的表达及沉默E盒锌指结合蛋白1(ZEB1)和细胞因子信号转导抑制因子1(SOCS1)水平的影响,探讨AFPV对SLE的作用及可能机制。结果:与正常组比较,模型组大鼠背部皮肤红肿直径显著增大(P 0. 01),与模型组比较,AFPV高、中剂量组可显著降低SLE大鼠背部皮肤红肿直径(P 0. 05,P 0. 01);与正常组比较,模型组大鼠血清Ig M,IL-6,IL-33明显增高(P 0. 05),与模型组大鼠比较,AFPV高、中给药组Ig M,IL-6,IL-33明显降低(P 0. 05);与正常组比较,MRL/lpr狼疮小鼠的CD4+T细胞miR-200a的表达显著降低,miR-155的表达显著增高及ZEB1水平显著增高而SOCS1的表达显著降低(P 0. 01),与模型组比较,AFPV干预后miR-200a的表达显著增高,miR-155的表达显著降低(P 0. 01),ZEB1水平显著降低而SOCS1的表达显著增高(P 0. 01)。结论:AFPV对SLE大鼠具有一定的治疗作用,其机制可能与其对miR-200a,miR-155及ZEB1和SOCS1的调节有关。     

多巴丝肼片对MTPT诱导C57BL/6J小鼠帕金森病模型的影响

[目的]建立MTPT诱导C57BL/6J小鼠帕金森病模型,探讨多巴丝肼片对MTPT诱导C57BL/6J小鼠帕金森病模型行为学变化的影响。[方法]选用纯系C57BL/6J雄性小鼠30只,按体重随机分为空白对照组、模型对照组及多巴丝肼片对照组,采用腹腔注射MTPT诱导C57BL小鼠帕金森病模型,使用多巴丝肼片灌胃进行预防给药,观察小鼠爬杆实验时间与悬挂实验评分的变化。[结果]经统计学处理数据表明,与模型对照组相比,多巴丝肼片可以明显降低小鼠爬杆实验时间(P<0.01)与明显提高悬挂实验评分(P<0.05)。[结论]多巴丝肼片对MTPT诱导C57BL/6J小鼠帕金森病模型有预保护防作用。以上结果为多巴丝肼片临床应用防治帕金森病提供了部分药效学实验依据。     

铁皮石斛多糖促进毛发生长的实验研究

目的：观察铁皮石斛多糖（DCP）促毛发生长的功效并探讨其作用机制,为铁皮石斛的开发利用奠定基础。方法：采用水提醇沉法提取DCP,并用苯酚-硫酸法测定其多糖含量。取C57BL/6J小鼠30只,用脱毛膏去除其背部毛发建立脱发模型,分为对照组、阳性对照组和DCP组,每日分别涂抹超纯水、米诺地尔酊、DCP（5.0 g·L^-1）l 次,每次 0.2 mL,连续给药21 d,应用小鼠毛发生长情况评分标准对C57BL/6J小鼠给药7,14 d毛发生长情况进行评分,及称量给药21 d C57BL/6J小鼠单位脱毛区域毛发质量评价DCP对C57BL/6J小鼠毛发生长的影响。采用MTT法和RT-PCR法分别评价DCP对人永生化角质形成细胞（HaCaT细胞）增殖和对HaCaT细胞血管内皮生长因子（VEGF）mRNA表达的影响。结果：DCP提取率为29.87%,DCP质量分数为79.65%。DCP组C57BL/6J小鼠的毛发生长平均得分、平均质量均显著优于对照组;DCP组HaCaT细胞生存率和VEGF mRNA的表达水平与对照组比较均显著提高。结论：DCP具有促毛发生长的作用,其机制可能是上调VEGF mRNA的表达。     

Nigella sativa modulates splenocyte proliferation, Th1/Th2 cytokine profile, macrophage function and NK anti-tumor activity

Aim of the study

Nigella sativa, also known as blackseed, has long been used in traditional medicine for treating various conditions related to the respiratory and gastrointestinal systems as well as different types of cancers. In this study, the potential immunomodulatory effects of Nigella sativa are investigated in light of splenocyte proliferation, macrophage function, and NK anti-tumor activity using BLAB/c and C57/BL6 primary cells.

Materials and methods

Splenocyte proliferation was assessed by [³H]-thymidine incorporation. Griess assay was performed to evaluate NO production by macrophages. ELISA was performed to measure the level of cytokines secreted by splenocytes and macrophages. NK cytotoxic activity against YAC-1 tumor cells was examined by JAM assay.

Results

We demonstrate that the aqueous extract of Nigella sativa significantly enhances splenocyte proliferation in a dose-responsive manner. In addition, the aqueous extract of Nigella sativa favors the secretion of Th2, versus Th1, cytokines by splenocytes. The secretion of IL-6, TNFα, and NO; key pro-inflammatory mediators, by primary macrophages is significantly suppressed by the aqueous extract of Nigella sativa, indicating that Nigella sativa exerts anti-inflammatory effects in vitro. Finally, experimental evidence indicates that the aqueous extract of Nigella sativa significantly enhances NK cytotoxic activity against YAC-1 tumor cells, suggesting that the documented anti-tumor effects of Nigella sativa may be, at least in part, attributed to its ability to serve as a stimulant of NK anti-tumor activity.

Conclusions

Our data present Nigella sativa as a traditionally used herb with potent immunomodulatory, anti-inflammatory, and anti-tumor effects. We anticipate that Nigella sativa ingredients may be employed as effective therapeutic agents in the regulation of diverse immune reactions implicated in various conditions and diseases such as cancer.     

粉防已碱对血管紧张素II诱导心肌肥大及p-ERK1/2表达的影响

目的：观察粉防已碱(tetrandrine, Tet)对血管紧张素II(angiotensin Ⅱ, Ang Ⅱ)诱导的心肌肥大及p-ERK_1/2表达及活性的影响。方法：培养新生大鼠心肌细胞,用相差显微镜计数心肌细胞搏动频率、细胞图像分析系统测量细胞体积、考马斯亮蓝法测定心肌细胞总蛋白含量、［₃H］-亮氨酸掺入法测定蛋白合成速率作为心肌肥大指标；以ERK免疫沉淀活性试剂盒测定ERK活性,Western-blot测定p-ERK_1/2表达。结果：Tet能显著抑制Ang II诱导的心肌细胞搏动频率、体积、总蛋白含量、蛋白合成速率的上升；对p-ERK_1/2表达及活性具有剂量依赖性的抑制作用。结论：Tet可以明显抑制Ang II诱导的心肌肥大,机制与降低p-ERK_1/2表达及活性有关。     

Vasodilatory Effects of Cinnamic Acid via the Nitric Oxide–cGMP–PKG Pathway in Rat Thoracic Aorta

Cinnamic acid (CA) and its derivatives have a broad therapeutic spectrum that includes antimicrobial, antifungal, and antitumoral activities. However, the vasodilative effect of CA has not been demonstrated. The present study characterizes the vasodilative activity and the mechanism of CA in rat thoracic aorta. The vasomotion of aortic strips following CA treatment was measured in an organ bath system. In addition, vascular strips and human umbilical vein endothelial cells (HUVECs) were used in organ bath, Western blot, nitrite, and cyclic guanosine monophosphate (cGMP) measurements. CA relaxed phenylephrine‐precontracted aortic strips in an endothelium‐dependent manner. Pretreatment of the endothelium‐intact aortic strips with N^G‐nitro‐l ‐arginine methyl ester (10^?4 M), 1 H‐[1,2,4]‐oxadiazolole‐[4,3‐a] quinoxalin‐10‐one, (10^?6 M) and methylene blue (10^?5 M) inhibited CA‐induced vasorelaxation. CA also increased the phosphorylation of endothelial nitric oxide synthase and nitric oxide generation in a concentration‐dependent manner in HUVECs. In addition, cGMP generation and cGMP‐dependent protein kinase G (PKG) expression in aortic strips were increased by CA treatment. Furthermore, CA‐induced vasorelaxation was inhibited by the PKG inhibitor KT5823 (0.3 μM) and the Ca²⁺‐activated K⁺ channel inhibitor tetraethylammonium (10^?3 M). These findings suggest that CA exerts an endothelium‐dependent vasodilation effect via the nitric oxide–cGMP–PKG‐mediated pathway in rat thoracic aorta. Copyright © 2012 John Wiley & Sons, Ltd.     

Cytotoxicity Effects and Apoptosis Induction by Bisbenzylisoquinoline Alkaloids from Triclisia subcordata

Triclisia subcordata Oliv (Menispermeaceae) is a medicinal plant traditionally used for the treatment of various diseases in West Africa. The ethanol extract of T. subcordata and its fractions were screened for in vitro anti‐ovarian cancer activities using the Sulforhodamine B assay. The crude alkaloids showed the strongest activity in cell growth assays on Ovcar‐8 and A2780 cell lines (IC₅₀ < 2.4 µg/mL). A bisbenzylisoquinoline alkaloid‐cycleanine was isolated using HPLC and identified by mass spectrometry and nuclear magnetic resonance analyses. The IC₅₀ values of cycleanine and tetrandrine (an alkaloid previously reported from this plant) ranged from 7 to 14 μM on Ovcar‐8, A2780, Ovcar‐4, and Igrov‐1 ovarian cancer cell lines. The IC₅₀ of cycleanine on human normal ovarian surface epithelial cells was 35 ± 1 μM, hinting at modest selectivity toward cancer cells. Both cycleanine and tetrandrine caused apoptosis as shown by activation of caspases 3/7 and cleavage of poly(ADP‐ribose) polymerase to form poly(ADP‐ribose) polymerase‐1 by using western blot analysis. Flow cytometry analyses showed that the percentages of apoptotic cells and cells in subG₁ phase increased after exposure of cycleanine and tetrandrine to Ovcar‐8 cells for 48 h compared with control. Cycleanine, like its isomer tetrandrine isolated from T. subcordata, could be a potential new anti‐ovarian cancer agent acting through the apoptosis pathway. Copyright © 2016 John Wiley & Sons, Ltd.     

A Mineral Extract from red Algae Ameliorates Chronic Spontaneous Colitis in IL‐10 Deficient Mice in a Mouse Strain Dependent Manner

Inflammatory bowel disease is an urgent public health problem with a high incidence in developed countries. Alterations of lifestyle or dietary interventions may attenuate the disease progression and increase the efficacy of current therapies. Here we tested the effect of chronic supplementation with a mineral extract from red marine algae – rich in calcium (34%), magnesium, phosphorus, selenium and other trace minerals – in a clinically relevant model of spontaneous enterocolitis, interleukin (IL)‐10^‐/‐ mice. The mineral extract was administered in the drinking water of Il10^‐/‐ mice on C57BL/6 J and BALB/c strain backgrounds for 25 weeks commencing from 3 to 4 weeks of age. The mineral extract ameliorated the spontaneous development of colitis and severity of disease in Il10^‐/‐ mice on a C57BL/6 J background. Mineral extract‐treated Il10^‐/‐ C57BL/6 J strain mice had significantly reduced mortality, circulating levels of serum Amyloid A and reduced colonic tissue damage. In contrast, comparable treatment of Il10^‐/‐ mice on a BALB/c background with the mineral extract did not alter the course of colitis. These data demonstrate that chronic supplementation with a natural mineral extract selectively ameliorates spontaneous mild–moderate colitis in Il10^‐/‐ mice on a C57BL/6 J, but does not attenuate more moderate–severe colitis in BALB/c strain animals. Copyright © 2013 John Wiley & Sons, Ltd.     

应用m2-Gi1α融合蛋白鉴别M2受体的特异性药物

 目的表达m2-Gilα融合蛋白,通过检测GDP与m2-Gilα融合蛋白的亲和力大小来鉴别m2受体的特异性激动剂、拮抗剂和反向激动剂。方法用两步PCR反应重组m2-Gilα融合蛋白cDNAs,并插入到pBacPAK9病毒载体中,利用Sf9细胞表达M2受体蛋白和m2-Gilα融合蛋白,通过[³H]QNB结合饱和实验及[³⁵S]GTPγS竞争性替代结合实验,检测受体表达水平及GDP 与m2-Gilα融合蛋白的亲和力。结果m2-Gilα融合蛋白表达水平是(18.14±0.17)pmol·mg(膜蛋白)^-1。M2-Gilα融合蛋白与[³H]QNB的结合量随着[³H]QNB浓度的升高而增加,10μmol·L^-1GTDγS使m2-Gilα融合蛋白与[³H]QNB的饱和结合曲线明显下移。Ach使m2-Gilα融合蛋白与[³⁵S]GTPγS竞争性替代结合曲线明显右移,Pilo稍次之,alcuronium,AF—DX116,Iso及atropine 则使曲线左移,GDP的IC₅₀值分别是168.4,152.3,6.84,6.12,5.59,4.52μmol·L^-1,无配体存在时为14.7μmol·L^-1。结论表达的m2-Gilα融合蛋白具备M2配体结合的特性及G蛋白耦联受体的信号转导功能,对于有基础活性的m2-Gilα融合蛋白,Ach, Pilo是完全激动剂;atropine,alcuronium,Iso和AF-DX116为拮抗剂。     

越鞠丸对Balb/c 小鼠和C57BL/6J 小鼠抗抑郁作用与前额叶场电位差异性分析

目的：比较Balb/c小鼠和C57BL/6J小鼠中越鞠丸抗抑郁作用的差异并分析其对小鼠前额皮层（prefontal cortex,PFC）突触传递的影响。方法：成年健康的雄性Balb/c小鼠和C57BL/6J小鼠随机分为对照组和越鞠丸组,给药30 min后进行强迫游泳测试（forced swimming test,FST）,运用电生理实验（electrophysiologicalexperiment）检测小鼠前额皮层的场电位（field excitatory postsynaptic potential,fEPSP）及长时程增强（long termpotentiation,LTP）。结果：强迫游泳测试中,Balb/c小鼠的越鞠丸组不动时间较对照组明显降低（P < 0.01）,C57BL/6J小鼠的的越鞠丸组和对照组在不动时间上没有明显差异（P > 0.05）。电生理实验中,Balb/c小鼠的越鞠丸组与对照组相比,场电位斜率百分比显著升高（P < 0.01）,而C57BL/6J小鼠的越鞠丸组和对照组的场电位斜率百分比无显著性差异（P > 0.05）;Balb/c小鼠的越鞠丸组与对照组相比,长时程增强显著升高（P < 0.01）,而C57BL/6J小鼠的越鞠丸组和对照组的长时程增强无显著性差异（P > 0.05）。结论：越鞠丸可能通过增加Balb/c小鼠前额皮层的场电位及长时程增强以提高突触传递效能,从而产生抗抑郁作用。     

Stimulating actions on ribonucleic acid biosynthesis of aconitines,diterpenic alkaloids of Aconitum roots

Mesaconitine (MA) significantly stimulated the incorporation of 5-[³H]orotic acid into liver nuclear RNA 16h after its administration. MA exhibited the strongest activity among the aconitine alkaloids. This stimulatory effect of MA was inhibited by actinomycin D, but the incorporation ratio of 5-[³H]orotic acid in the combined treatment group with MA and actinomycin D was significantly higher than that in the single treatment group with actinomycin D. MA also increased the incorporation of 5-[³H]orotic acid into polysomal RNA in mouse liver. When MA was added to the RNA polymerase (EC 2.7.7.6) preparation obtained from rat liver, the increase of the enzyme activity was weak. While RNA polymerase preparation from the liver of rats, which were previously treated with MA, elicited increased incorporation of [³H]cytidine monophosphate into RNA as compared with that from the livers of normal rats.Accumulated data indicate that aconitine alkaloids, in particular MA, accelerate liver RNA synthesis mainly by the increase of RNA polymerase.     

In vitro study of the interaction of Tilia europeae L. aqueous extract with GABAA receptors in rat brain

The interaction of GABA_A receptor-complex in rat brain was investigated in vitro with aqueous extracts obtained from the inflorescences of Tilia europeae L., using the [³H]muscimol and [³H]flunitrazepam binding techniques to synaptic membranes and the uptake of ³⁶Cl^- to synaptoneurosomes from cortices. The extract inhibited [³H]muscimol binding, stimulated ³⁶ClCl^- uptake by synaptoneurosomes and displaced at high concentrations, the [³H]flunitrazepam bound to synaptic membranes. When analysed by HPLC, the aqueous extract of Tilia europeae L. contained several amino acids, including GABA (about 100 μM ). This GABA content can justify the displacement of [³H]muscimol produced by the extract but it did not increase the binding of [³H]flunitrazepam, as expected. Probably the extract contains other benzodiazepine-like substances which displace the [³H]flunitrazepam binding and counteract the expected GABA-induced increase in [³H]flunitrazepam binding. © 1997 John Wiley & Sons, Ltd.     

Effects of the standardized Panax ginseng extract g115 on the D-glucose transport by Ehrlich ascites tumour cells

The effect of standardized Panax ginseng extract (G115 Pharmaton) on D -glucose uptake by Ehrlich ascites tumour cells from mouse was examined. Measurements were carried out using [³H]-2-deoxy-D -glucose, a non-metabolizable glucose analogue. [³H]-2-deoxy-D -glucose was transported from the medium to cells and phosphorylated but was neither further metabolized nor eliminated. Thus, the amount taken in the cells was a measure of D -glucose uptake. The results showed that G115 stimulated D -glucose transport and the maximum effect was obtained at a G115 concentration of 2.0 μg/mL representing an increase of about 35% above basal activity.     

重组人5型腺病毒对B16黑色素瘤生长的抑制作用

 目的研究瘤内直接注射重组人5型腺病毒H101对B16黑色素瘤生长以及对手术后再次接种B16黑色素瘤细胞后肿瘤生长的影响。方法建立小鼠B16黑色素瘤动物模型。瘤内直接注射H101腺病毒,测量肿瘤体积,切除肿瘤,再次接种B16黑色素瘤细胞,观察肿瘤生长情况。结果细胞接种8 d后,全部小鼠均长出肿瘤。H101注射组肿瘤的体积在给药后3,6,9 d,都显著小于对照组。再次接种B16黑色素瘤细胞后,H101注射组肿瘤的生长速度也显著小于对照组。结论瘤内直接注射H101对B16黑色素瘤生长具有抑制作用,对手术后再次接种的肿瘤生长也具有抑制作用。     

麦考酚酸体外对肝脏自然杀伤细胞活性及其受体表达的影响

 目的探讨麦考酚酸(MPA)对肝脏自然杀伤(NK)细胞活性的影响及其机制。方法分离C57BL/6小鼠肝脏NK细胞,用不同浓度MPA处理24h后,³H-TdR释放法检测NK细胞杀伤活性,ELISA法检测培养上清IFN-γ,IL-2,IL-10含量,流式细胞术检测NK细胞受体NKG2D,NKG2A,Ly49A的表达情况。结果不同浓度MPA处理后肝脏NK细胞杀伤活性较正常对照明显降低(P<0.01),IFN-γ,IL-2分泌量降低,而IL-10分泌量增高,NK细胞活化性受体NKG2D和抑制性受体Ly49A的表达下调,抑制性受体NKG2A的表达上调,高浓度的MPA作用后结果更明显。结论MPA通过下调肝脏NK细胞活化性受体NKG2D并上调抑制性受体NKG2A而抑制肝脏NK细胞活性并影响细胞因子的分泌。