核磁共振法测定熊去氧胆酸中鹅去氧胆酸的含量

用一般的滴定方法测定熊去氧胆酸中鹅去氧胆酸的含量是很难进行的。NMR方法能有效地进行测定。样品(10～20毫克)溶解在CDCl_3和DMSO-d_6(8:1v/v)的混合溶剂中,马来酸做标准物,以δ=3.8ppm的鹅去氧胆酸C_7质子峰面积计算含量。方法的变异系数是0.45-1.3%,回收率是99.3-100.1%。     

薄层荧光扫描法测定利肝片中猪去氧胆酸及鹅去氧胆酸含量

目的：建立利肝片中猪去氧胆酸(HDCA)及鹅去氧胆酸(CDCA)的含量测定方法。方法：在硅胶G板上以环己烷-醋酸乙酯-醋酸-甲醇(20：25：2：3)展开，以薄层荧光扫描法，激发波长为366nm,测定利肝片中HDCA及CDCA的含量。结果：平均回收率分别为HDCA：99．9％，RSD=1．9％；CDCA：100.9％，RSD=2．8％。结论：该方法灵敏，准确，适于作为该制剂的质控标准。     

鹅去氧胆酸制备新工艺

用氯化钙取代老工艺中所用的氧化钡,利用鹅去氧胆酸钙盐与杂质钙盐在水中溶解度的不同,分离纯化了鹅去氧胆酸,收率3.2％。     

胆酸止咳片中猪去氧胆酸和鹅去氧胆酸的含量测定

目的 建立高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法测定胆酸止咳片中猪去氧胆酸和鹅去氧胆酸的含量.方法 采用Agilent SB C18色谱柱(250 mm×4.6 mm,5 μm),柱温30 ℃,流动相为甲醇-乙腈-0.1%甲酸溶液(62: 14: 24),流速0.8 mL.min-1,检测器为ELSD,漂移管温度为85 ℃,氮气流速为2.3 L.min-1.结果 猪去氧胆酸含量在0.427 0～6.405 0 mg范围内有良好线性关系,平均加样回收率为99.7%,RSD＝1.9%(n＝9);鹅去氧胆酸含量在0.218 6～3.279 0 mg范围内有良好线性关系,平均回收率为97.5%,RSD＝1.2%(n＝9).结论 该方法 简便,结果 准确,重复性好,可用于胆酸止咳片的质量控制.     

猪去氧胆酸合成熊去氧胆酸的研究进展

临床上用于治疗肝胆疾病的熊去氧胆酸最初是从熊的胆汁中提取得到,受熊胆汁来源的限制目前主要是以胆酸为原料合成而得。之后随着鹅去氧胆酸提取工艺和鹅去氧胆酸合成熊去氧胆酸工艺的成熟,鹅去氧胆酸逐渐代替胆酸成为生产熊去氧胆酸的主要原料。由于猪去氧胆酸来源丰富、价格低廉,近几年由猪去氧胆酸合成熊去氧胆酸的研究成为甾类药物合成研究的热点之一。本文对这一领域的研究进展进行综述,以期为相关研究提供有价值的参考。     

HPLC-ELSD法同时测定护肝片中猪去氧胆酸和鹅去氧胆酸的含量

目的：建立高效液相-蒸发光散射检测（ HPLC-ELSD）法同时测定护肝片中猪去氧胆酸和鹅去氧胆酸含量的方法。方法采用Agilent Zorbax SB-C18（4.6 mm×250 mm,5μm）色谱柱,流动相为甲醇-0.2%冰醋酸溶液（80：20）,流速为1.0 mL·min-1,色谱柱温为30℃;蒸发光检测器载气为氮气,流速为2.0 L·min-1,漂移管温度为60℃。结果猪去氧胆酸在0.521~10.42μg范围内呈现良好的线性关系（ r＝0.9989）,平均加样回收率为97.41%, RSD为1.30%（n＝6）;鹅去氧胆酸在0.266~5.32μg范围内呈现良好的线性关系（r＝0.9994）,平均加样回收率为96.09%,RSD为2.11%（ n＝6）。结论该方法准确、简便、灵敏度高,可用于护肝片中猪胆粉的质量控制。     

延龄草苷对脂多糖刺激的小胶质细胞炎症抑制作用

目的 探讨延龄草苷（trillin）对小胶质细胞（BV-2）的炎性激活的抑制作用。方法 不同浓度的延龄草苷孵育0.5 h,然后用脂多糖进行刺激,培养24 h后收集上清,Greiss法检测NO浓度;CCK-8检测延龄草苷对小胶质细胞活力的影响;Elisa检测肿瘤坏死因子（TNF-α）的浓度;实时定量PCR（RT-PCR）检测TNF-α、IL-1β、IL-6、iNOS、COX-1、COX-2基因的表达。结果 在不影响细胞活力的前提下,延龄草苷能明显抑制NO的释放,同时能抑制激活诱导细胞死亡,能明显抑制TNF-α的产生,能明显抑制TNF-α、IL-1β、IL-6、iNOS、COX-2 mRNA的表达,但是不能抑制COX-1 mRNA的表达。结论 延龄草苷可以抑制LPS诱导的小胶质细胞的炎性激活,抑制炎性基因的表达。     

Inhibitory effects of eicosapentaenoic acid on lipopolysaccharide-induced activation in BV2 microglia

Upon activation, microglia release proinflammatory mediators that play important roles in eliciting neuroinflammatory responses associated with neurodegenerative diseases. The anti-inflammatory properties of eicosapentaenoic acid (EPA) have been known, however, the effects responsible for lipopolysaccharide (LPS)-induced activation remain poorly understood in microglia. In the present study, we investigated the effects of EPA on the expression of proinflammatory mediators in LPS-stimulated BV2 microglia. EPA significantly inhibited the release of nitric oxide (NO), prostaglandin E(2) (PGE(2)) and proinflammatory cytokines such as interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha in a dose-dependent manner. EPA also attenuated the production of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and proinflammatory cytokines at mRNA and/or protein levels. Moreover, EPA suppressed NF-kappaB activation by blocking IkappaB degradation, and also blocked the mitogen-activated protein kinases (MAPKs) such as ERK, p38 and JNK, and the Akt pathway. The anti-inflammatory properties of EPA may be useful for ameliorating neurodegenerative diseases as well as suppressing LPS-induced shock.     

红景天苷调控PI3K/Akt信号通路对LPS诱导的BV2小胶质细胞的抗炎作用

目的探讨红景天苷对脂多糖(LPS)诱导的BV2小胶质细胞的抗炎作用及其机制。方法建立LPS诱导BV2小胶质细胞损伤模型。经不同浓度的红景天苷作用后,q PCR法检测细胞因子IL-6、IL-1β、TNF-αmRNA的表达; Western blot法检测Akt、p-Akt、核蛋白NF-κB p50的蛋白表达。PI3K抑制剂LY294002作用30 min,再经红景天苷作用后,检测Akt、p-Akt、核蛋白NF-κB p50、IL-6、IL-1β、TNF-α等指标。结果与模型组比较,红景天苷能够抑制BV2小胶质细胞IL-6、IL-1β、TNF-αmRNA的表达,促进p-Akt蛋白表达,抑制核蛋白NF-κB p50;经LY294002作用后,红景天苷对pAkt、核蛋白NF-κB p50、IL-6、IL-1β等作用不明显。结论红景天苷能够抑制LPS诱导的BV2小胶质细胞炎症反应,主要是通过激活PI3K/Akt信号通路,促进Akt的磷酸化,抑制NF-κB p50核转录,进而抑制细胞因子。     

槟榔碱改善脂多糖诱导的BV2细胞神经炎症及作用机制

目的 探讨槟榔碱对脂多糖（LPS）诱导小鼠小胶质细胞BV2炎症反应的改善作用及机制。方法 取对数生长期的BV2细胞分为空白组和LPS(0.01、0.1、1、10、20μg/mL)组,CCK-8法检测细胞活力,分光光度法检测一氧化氮（NO）含量。另取细胞分为空白组、模型组、槟榔碱（10、20、40μmol/L）组。CCK-8法检测细胞活力;分光光度法检测NO含量;酶联免疫吸附法（ELISA）检测上清中肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)和抗炎因子白细胞介素10(IL-10)水平;实时荧光定量PCR(qPCR)法检测细胞中TNF-α、IL-6、IL-1β、一氧化氮合酶（i NOS）mRNA表达表达;Western blotting法检测Toll样受体4(TLR4)、p-p65、p65、环氧化酶2(COX2)、iNOS、胞内磷脂酰肌醇激酶（PI3K）、蛋白激酶B(Akt)、p-Akt蛋白表达水平。结果 0.01～20μg/mL LPS诱导对BV2小胶质细胞的细胞活力无显著影响,1μg/mL LPS诱导显著上调了细胞中NO含量;槟榔碱在10～4...     

二氯乙酸钠对氧糖剥夺损伤的BV2细胞的保护作用及其机制研究

目的 研究二氯乙酸钠（dichloroacetate,DCA）对氧糖剥夺（oxygen glucose deprivation,OGD）损伤模型中小鼠小胶质细胞（BV2细胞）的保护作用,并探讨其作用机制。方法 将BV2细胞分为3组：对照组、OGD组、DCA治疗组,通过OGD 4 h建立损伤模型。CCK-8和流式细胞仪检测细胞凋亡及ROS和NO的表达,Western blot检测NF-κB通路相关蛋白表达水平。结果 CCK-8及流式细胞检测结果表明,DCA可显著降低OGD诱导的BV2细胞凋亡,并且减少OGD后细胞中活性氧（ROS）和一氧化氮（NO）的表达（P<0.05）。Western blot结果显示,DCA可显著影响OGD损伤介导的BV2细胞JNK、I-κB和NF-κB蛋白表达水平（P<0.05）。结论 DCA对OGD损伤的BV2细胞具有保护作用,其机制与抗凋亡、抗氧化和抗炎作用有关。     

Effects of baicalin and geniposide on polarization of BV2 microglia

OBJECTIVE To investigate the effects of baicalin(BC), geniposide(GP) and 7∶3 combination(BC/GP) on polarization of BV2 microglia. METHODS LPS(50 μg·L~(-1)) and IL-4(20 μg·L~(-1)) were polarized to M1 and M2. The optimal concentrations of BC, GP and BC/GP were detected by CCK-8, and then each group was divided into high, middle and low dose group in the optimal concentration range. The Griess method was used to detect the NO content in the supernatant of each group. The contents of IL-10, TGF-β1, IL-1β and TNF-α in the supernatant of each group were detected by ELISA. The cells in each group were labeled with fluorescent dye-labeled CD86 and CD206 antibodies and the fluorescence expression of M1 and M2 was observed by fluorescence microscope. RESULTS CCK-8 results showed that BC, GP and BC/GP had no significant effect on cell status from 32 to 125 μmol·L~(-1). The content of NO in LPS group was significantly higher than that in blank group(P<0.01), but there was no significant difference in IL-4 group compared with blank group, and each drug group was significantly lower than that in LPS group(P<0.01). Compared with LPS group, the contents of IL-10 and TGF-β1 were increased(P<0.05), and the contents of TNF-α and IL-1β were decreased in each drug group and IL-4 group(P<0.05). Fluorescence microscopy showed the fluorescence intensity of surface markers of M1 and M2 microglia CD86 and CD206. It was found that the expression of CD86 was weaker and the expression of CD206 was enhanced in each of the administration group and IL-4 group. CONCLUSION A certain concentration of LPS and IL-4 can polarize BV2 microglia into M1 and M2 type, BC, GP and their 7∶3 combination can reduce inflammatory factors, promote the release of anti-inflammatory factors and drive microglia to M2 polarization and have a protective effect on nerve damage after cerebral ischemia.     

Anti-inflammatory effects of 5-aminosalicylic acid conjugates with chenodeoxycholic acid and ursodeoxycholic acid on carrageenan-induced colitis in guinea-pigs.

Two epimeric bile acid conjugates, 5-aminosalicylic acid-chenodeoxycholic acid (5-ASA-CDCA) and 5-aminosalicylic acid-ursodeoxycholic acid (5-ASA-UDCA), were synthesized to deliver 5-ASA to the large intestine by oral administration. The movement of the conjugates down the gastrointestinal tract and the anti-inflammatory effects on ulcerative colitis were investigated by administering the conjugates to guinea-pigs with an inflammatory bowel disease induced by 2% degraded carrageenan solution. The conjugates were protected from deconjugation in stomach and small intestine and reached the caecum and the colon, where 5-ASA was more easily liberated from 5-ASA-CDCA than from 5-ASA-UDCA. The conjugates at doses equivalent to 50 or 150 mg kg(-1) 5-ASA were orally administered once a day for 4 weeks from the 15th day after starting carrageenan treatment. The body weights and the bleeding scores of occult blood in faeces were measured during the experiment. The number of ulcers in the caecum and the colon were counted after killing the guinea-pigs at the end of the experiment. Rapid onset of efficacy was shown by a significant reduction in bleeding scores within a week after administration of the conjugates. Treatment with the lower dose of 5-ASA-CDCA showed a recovery of body weight and a significantly decreased number of ulcers in the caecum, and the ulcers in the colon had completely disappeared bythe end of the experiment. There was a good correlation found between the number of ulcers in the caecum and the bleeding scores of occult blood in faeces. The findings indicate that both conjugates were sufficiently delivered to the large intestine without deconjugation and that the lower dose of 5-ASA-CDCA is enough for treatment of ulcerative colitis in colonic inflammatory bowel diseases.