消退素D1抑制NLRP3信号通路对DSS结肠炎小鼠的影响

目的探讨消退素D1(resolvin D1,RvD1)对实验性结肠炎小鼠肠道炎症的影响及可能机制。方法将C57BL/6小鼠分为4组,包括正常组、RvD1对照组、葡聚糖硫酸钠(dextran sodium sulfate, DSS)模型组、RvD1治疗组。实验结束后,分离小鼠小肠及结肠组织。测量疾病活动指数(DAI),病理学评分(HI),检测髓过氧化物酶(MPO)活性水平,伊文思蓝检测肠黏膜通透性,电镜检测肠上皮细胞连接及细胞因子水平。分析NLRP3炎症小体相关基因表达水平。结果与正常组小鼠相比,模型组DAI评分、HI评分、MPO活性水平,以及结肠组织匀浆中促炎细胞因子的产生明显增加(P<0.05)。RvD1组小鼠上述实验指标明显减低(P<0.05)。模型组小鼠结肠组织NLRP3炎症小体表达水平增加(P<0.05);RvD1处理1周,小鼠结肠组织NLRP3通路相关蛋白表达水平降低(P<0.05)。结论 RvD1具有改善DSS结肠炎小鼠肠道炎症的作用,其机制可能与抑制NLRP3炎性体信号通路有关。     

Ginsenoside Rg1 attenuates motor impairment and neuroinflammation in the MPTP-probenecid-induced parkinsonism mouse model

OBJECTIVE To evaluate these activities of Rg1 in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)/probenecid(MPTP/p)-induced PD mouse model for the first time and to elucidate the underlying mechanisms.METHODS Male C57BL/6 mice were randomly assigned to six groups.One hour prior to MPTP/p injection,GroupⅢ-Ⅵmice received 10 mg·kg~(-1),20 mg·kg~(-1),or 40 mg·kg~(-1) Rg1 or 3 mg·kg~(-1) selegiline,respectively,orally from D(-3) to D49.GroupⅠ-Ⅱmice received solvent water.Subsequently,GroupⅡ-Ⅵmice received by injection MPTP-HCl(25 mg·kg~(-1) bw dissolved in0.9%saline,sc)on a 40-d schedule at intervals of 4 d between consecutive doses in combination with an adjuvant drug,probenecid(250 mg·kg~(-1) bw in 0.03 mL of DMSO,ip);GroupⅠmice were injected with saline and probenecid.Behavioral performance was assessed in the open field test,pole test and rotarod test.Neurotransmitters in the striatum were detected using HPLC.Protein levels were measured by Western blot.Pathological characteristics were examined by immunohistochemistry.Ultrastructure changes were observed by electron microscopy.RESULTS Oral treatment with Rg1 significantly attenuated the high MPTP-induced mortality,behavior defects,loss of dopamine neurons and abnormal ultrastructure changes in the SNpc.Other assays indicated that the protective effect of Rg1 may be mediated by its anti-neuroinflammatory properties.Rg1 regulated MPTP-induced reactive astrocytes and microglia and decreased the release of cytokines such as tumor necrosis factor-α(TNF-α)and interleukin-1b(IL~(-1)b)in the SNpc.Rg1 also al eviated the unusual MPTP induced increase in oligomeric,phosphorylated and disease-related a-synuclein in the SNpc.CONCLUSION Rg1 protects dopaminergic neurons,most likely by reducing aberrant a-synuclein-mediated neuroinflammation,and holds promise for Parkinson disease therapeutics.     

Ginsenoside Rd attenuates the inflammatory response via modulating p38 and JNK signaling pathways in rats with TNBS-induced relapsing colitis

In this study, we investigated the effects and the protective mechanism of ginsenoside Rd (GRd) which has been identified as one of the effective compounds from ginseng on relapsing colitis model induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) in rats. After inducing relapsing colitis in experimental rats on two occasions by intracolonic injection of TNBS, GRd (10, 20 and 40 mg/kg) was administered to experimental colitis rats for 7 days. The inflammatory degree was assessed by macroscopic score, histology and myeloperoxidase (MPO) activity. The levels of proinflammatory cytokines, such as TNF-α, IL-1β, and IL-6 were determined by ELISA. Mitogen-activated protein kinase (MAPK) phosphorylation was analyzed by western blotting method. The results showed that GRd markedly attenuates the inflammatory response to TNBS-induced relapsing colitis, as evidenced by improved signs, increased body weight, decreased colonic weight/length ratio, reduced colonic macroscopic and microscopic damage scores, inhibited the activity of MPO, lowered proinflammatory cytokine levels and suppressed phosphorylation of p38 and JNK. The possible mechanism of protection on experimental colitis after GRd administration was that it could reduce the accumulation of leukocytes and down-regulate multiple proinflammatory cytokines through modulation of JNK and p38 activation.     

Ginsenoside Rg1 prevent and treat inflammatory diseases: A review

Ginseng has been used as reinforcing drugs or for traditional Chinese medicine in aging, inflammation, stress, diabetes mellitus, hepatic diseases and cancer. The aim of this current review is to provide an integrative overview of the uses, relative human diseases and related mechanisms of ginseng. Nowadays, numerous animal experiments and clinical studies conducted to investigate the efficacy and safety of ginseng components. Inflammation is an immune response that protects human from pathogens, toxins and other dangers, which is initiated by recognizing pathogen- or danger- associated molecular patterns. Inflammasomes are cytosolic protein complexes which form in response to challenges, which also controls the activity of caspase-1, important for maturation and release of cytokines. Ice protease-activating factor, oligomerization domain-like receptor family pyrin domain-containing 1 and absent in melanoma 2 inflammasome recognize peculiar substances, while NLRP3 inflammasome responds towards structurally and chemically diverse triggers. The functional relationship between ginsenosides and inflammasome provides new insight into the understanding of molecular mechanisms of ginsenoside-mediated anti-inflammatory actions. It also has applications regarding the development of anti-inflammatory remedies by ginsenoside-mediated targeting inflammasomes, which could prevent and treat inflammatory diseases.     

Combined IMIG and immune Ig attenuates inflammatory colitis in mice

BackgroundUsing a combination of homologous and heterologous (mouse/human) polyclonal anti-idiotypic Igs and immune Igs in BALB/c mice we have previously reported attenuation of allergic type responses following OVA immunization. We have now investigated attenuation of an inflammatory colitis in C57BL/6 mice receiving dextran sodium sulfate (DSS) in their drinking water, using additional treatment of DSS-exposed mice with combined human Igs, commercial IVIG (given IM, hence hereafter IMIG) as a source of pooled anti-idiotype Ig, and human anti-Tet as immune Ig.MethodsAcute or chronic colitis was induced by DSS in groups of C57BL/6 mice. Mice also received weekly immunotherapy with im injections of polyclonal immune Ig, polyclonal anti-idiotype Ig, or the combined Igs, for a total of 5 injections, beginning with DSS treatment or after 2 cycles of DSS. Weight loss and mortality were monitored daily, and the extent of colitis was determined further using colonic length measurement, and by ELISA measurement of inflammatory cytokines in supernatants from colonic explant cultures.ResultsMice developed colitis in both the acute and chronic models with loss of body weight, shortened colon lengths, and increased expression of inflammatory cytokines in colonic tissue. Loss of body weight, and inflammatory cytokine production, was attenuated only in chronic colitis, and only after combined IMIG and immune Ig treatment, and not in groups receiving only IMIG or immune Ig alone.ConclusionHeterologous combinations of polyclonal IMIG and immune Ig can attenuate inflammatory colitis in mice. Given the described efficacy of this treatment for allergic desensitization, we hypothesize this methodology may have widespread clinic utility.     

Ginsenoside Rg3 attenuates ovariectomy-induced osteoporosis via AMPK/mTOR signaling pathway

Ginsenoside Rg3, a ginsenoside isolated from Panax ginseng, can regulate autophagy via AMP-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) signaling pathway. AMPK/mTOR signaling and autophagy have been reported to be involved in osteogenesis. Here, the effect of Rg3 on ovariectomy (OVX)-induced osteoporosis is explored. In vivo, rats were treated with 20 mg/kg Rg3 after OVX and the body weight (BW) was monitored. Bone mineral density (BMD), hematoxylin–eosin staining of femur tissues, osteogenesis, autophagy, and AMPK/mTOR signaling were analyzed. In vitro, MC3T3-E1 cells were treated with 0, 1, 5, 10, 20, and 100 μmol/L Rg3. 10 and 20 μmol/L Rg3, which had no significant effect on cell viability and significantly affected AMPK/mTOR signaling, were chosen for further analysis. Then osteogenic differentiation was induced with Rg3 or/and AMPK inhibitor (Compound C). AMPK/mTOR signaling, autophagy, osteogenic differentiation, and mineralization by Alizarin Red staining were analyzed. The expression or activity of AMPK/mTOR signaling-related proteins, autophagy markers, and osteogenesis markers was measured by western blotting or commercial kits, and cell viability by cell counting kit-8 assay kits. Rg3 significantly alleviated OVX-induced BW increases, BMD declines and histological changes of femur tissues, promoted osteogenesis, autophagy, and AMPK signaling, but inhibited mTOR signaling in vivo. Moreover, Rg3 significantly enhanced AMPK signaling, autophagy, osteogenic differentiation, and mineralization, but suppressed mTOR signaling in vitro. However, Compound C significantly reversed Rg3-induced alterations in vitro, indicating that Rg3 regulated autophagy, osteogenic differentiation, and mineralization via AMPK/mTOR signaling. Hence, it was speculated that Rg3 might attenuate OVX-induced osteoporosis via AMPK/mTOR signaling pathway.     

复方三七颗粒中人参皂苷Rg1的含量测定

目的：建立新方法测定复方三七颗粒中人参皂苷Rgl。方法：样品前处理方法改为甲醇提取后，用1％氢氧化钠溶液溶解，用大孔树脂吸附，用70％乙醇溶液洗脱，用TLC检测，回收率为98．4％，RSD=1．43％。结论：方法科学，结果准确。     

HPLC-ELSD法测定参银颗粒中人参皂苷Rg1及Re的含量

目的：用HPLC～ELSD法测定参银颗粒中人参皂苷Rgl及Re的含量。方法：色谱柱填料为十八烷基键合硅胶,流动相为乙腈-水;温度40℃,气体压力3．5bar。结果：人参皂苷Rgl及Re分别在0．9012～9．012,1．219—12．19μg之间呈良好的线性关系;参银颗粒中2种成分的平均回收率分别为98．2％（RSD1．3％）,97．8％（RSD1．5％）;结论：该方法简便、准确、分离效果好,无干扰,可用于参银颗粒的质量评价。     

健胃颗粒中人参皂甙Rg1的含量测定

健胃颗粒是在临床验方的基础上采用现代工艺研制成功的一种新制剂 ,由人参、黄连等七味药组成。具有健脾和胃 ,散结除痞之功效 ,用于治疗慢性萎缩性胃炎。为控制其质量 ,以方中主药人参中的人参皂甙Ｒｇ1为指标 ,采用薄层扫描法进行含量测定。1　仪器和试剂双波长薄层扫描仪 (日本岛津ＣＳ - 90 0 0型 ) ;硅胶Ｇ (青岛海洋化工厂 ) ;人参皂甙Ｒｇ1标准品(中国药品生物制品检定所提供 ) ;其它试剂均为分析纯。2　实验方法及结果2 1　扫描条件　反射法锯齿扫描 ,λｓ =5 2 5ｎｍ ,λＲ=70 0ｎｍ ;狭缝 :1 2 5ｎｍ× 1 2 5ｎｍ ,ＳＸ =3;灵敏…     

RP-HPLC法测定人参健脾片中人参皂苷Rg1、Re的含量

建立人参健脾片中测定人参皂苷Rg1、Re含量的反相高效液相色谱分析方法.采用Alltima-C18为分析柱,流动相为乙腈-0.05%磷酸溶液(20∶80);流速为1.0 mL·min-1;检测波长为203 nm.人参皂苷Rg1在41.24～1031μg·mL-1内与其峰面积呈良好的线性关系(r=0.9999),平均回收率为97.0%,RSD=2.2%(n=6).人参皂苷Re在17.248～431.2μg·mL-1内峰面积与浓度呈良好的线性关系(r=0.9997)平均回收率为96.4%,RSD=2.3%(n=6).该方法操作简便、快捷,结果准确,可用于人参健脾片的质量控制.     

HPLC法测定骨刺宁胶囊中人参皂苷Rg1的含量

目的：建立测定骨剌宁胶囊中人参皂苷Rg1的含量测定方法。方法：采用AichromBond-1C18（4．6mm×250mm,5μm）,以乙腈-水（20：80）为流动相;流速：1．0ml／min;柱温：25℃。检测波长：203nm。结果：人参皂苷Rg1在3．765-10．040μg范围内线性良好,回归方程为：Y=269005X-5893．7,r=0．9996,平均回收率97．33％,RSD为1．19％。结论：该法简便、准确、重现性好,可用于本制剂质量控制。     

HPLC法同时测定人参固本丸中人参皂苷Rg1和人参皂苷Re的含量

目的 建立了测定人参固本丸中的有效成分人参皂苷Rg1和人参皂苷Re的高效液相色谱测定方法.方法 采用UltimateTM XB-C18柱(250mm×4.6mm 5μm);乙腈-0.1%磷酸溶液(20∶80)为流动相;检测波长为203nm;流速1.0mL·min-1;柱温35℃.结果 人参皂苷Rg1在0.0752μg~1.8800μg具有良好的线性关系(r=0.9996),平均回收率为97.8%,RSD为0.011%(n=9);人参皂苷Re在0.1887μg~4.7175μg的范围具有良好的线性关系(r=0.9992),平均回收率为98.2%,RSD为0.015%(n=9).结论 本法快速、准确,可同时测定人参固本丸中人参皂苷Rg1、人参皂苷Re的含量.     

高效液相色谱法同时测定姜参胶囊中人参皂苷Rg_1、Re的含量

目的:建立以高效液相色谱法同时测定姜参胶囊中人参皂苷Rg1、Re含量的方法。方法:色谱柱为Kromasil C18,流动相为乙腈-0.05%磷酸溶液(21∶79),检测波长为203nm,流速为1.0mL.min-1。结果:人参皂苷Rg1、Re进样量分别在0.502~4.016μg(r=0.9997)、1.090~8.720μg范围内呈良好的线性关系(r=0.9998);平均回收率分别为98.8%(RSD=1.24%)、99.4%(RSD=1.68%)。结论:本法操作简便、快速、准确可靠,可用于姜参胶囊的质量控制。     

HPLC法测定参雄温阳胶囊中人参皂苷Rg1、Re的含量

目的：建立高效液相色谱法同时测定参雄温阳胶囊中人参的有效成分人参皂苷Rg。和人参皂苷Re的含量。方法：采用Spherisorb C18柱（250mm×4．6mm,5μm）;乙腈-0．1％磷酸溶液（20：80）为流动相;检测波长为203nm;流速1．0ml·min-1;柱温35℃。结果：人参皂苷Rg1在0．59—11．82μg具有良好的线性关系（r=1．0000）,平均回收率为99．8％,RSD为1．01％（n=9）;人参皂苷Re在1．04—20．74μg的范围具有良好的线性关系（r=0．9999）,平均回收率为98．7％,RSD为1．35％（n=9）。结论：本法快速、准确,可同时测定参雄温阳胶囊中人参皂苷Rg1、人参皂苷Re的含量。     

Calea uniflora Less. attenuates the inflammatory response to carrageenan-induced pleurisy in mice

Calea uniflora Less. (family Asteraceae), also named “arnica” and “erva-de-lagarto”, is a native plant to the South and Southeast of Brazil. This species was used to treat rheumatism, respiratory diseases, and digestive problems in Brazilian folk medicine. In vitro studies have shown the important biological effects of C. uniflora. However no studies have focused on the mechanism of action of anti-inflammatory activity of C. uniflora. The aim of this study was to evaluate the anti-inflammatory effects of the crude extract, its fractions, and isolated compounds obtained from of C. uniflora, using mouse model of carrageenan-induced inflammation. The following inflammatory parameters: leukocyte influx, degree of exudation, myeloperoxidase (MPO) and adenosine deaminase (ADA) activities, nitric oxide metabolites (NOx), proinflammatory cytokines and phosphorylation of the p65 subunit of NF-κB (p-p65 NF-κB), and p38 mitogen-activated protein kinase (p-p38 MAPK) levels were determined. The crude extract of C. uniflora, its fractions and its isolated compounds reduced the leukocyte influx, degree of exudation, MPO and ADA activities, NOx, TNF-α, IFN-γ, MCP-1 and IL-6 levels (p < 0.05). The isolated compounds reduced p-p65 NF-κB and p-p38 MAPK levels (p < 0.01). This study demonstrated that C. uniflora exhibits a significant anti-inflammatory activity via inhibition of the leukocyte influx and degree of exudation. These effects were associated with a decrease in the levels of several proinflammatory mediators. The mechanism of the anti-inflammatory action of C. uniflora may be, at least in part, via the inhibition of p65 NF-κB and p38 MAPK activation by the isolated compounds.     

Croton antisyphiliticus Mart. attenuates the inflammatory response to carrageenan-induced pleurisy in mice

The aim of this study was to investigate the anti-inflammatory effect of the crude hydroalcoholic extract (CHE) from the aerial parts of Croton antisyphiliticus, its fractions and isolated compounds derived from it on the mouse model of pleurisy induced by carrageenan. The aerial parts of C. antisyphiliticus were dried, macerated and extracted with ethanol to obtain the CHE, which was fractionated by liquid–liquid extraction using solvents with increasing polarity to obtain hexane (Hex), ethyl acetate (EA) and aqueous (Aq) fractions. Vitexin and quinic acid were isolated from Aq fraction. Capillary electrophoresis analysis, physical characteristics and spectral data produced by infrared (IR), nuclear magnetic resonance (¹H and ¹³C NMR) and mass spectrometry analyses were used to identify and elucidate the structure of the isolated compounds. The experimental model of pleurisy was induced in mice by a single intrapleural injection of carrageenan (1 %). Leukocytes, exudate concentrations, myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities and nitrate/nitrite (NOx), tumor necrosis factor-α (TNF-α) and interleukin-17 (IL-17) levels were determined in the pleural fluid leakage at 4 h after pleurisy induction. Animals pre-treated with CHE, Hex, EA, Aq, vitexin and quinic acid exhibited decreases in leukocytes, exudate concentrations, MPO and ADA activities and NOx levels (p < 0.05). Also CHE, Hex, EA and vitexin but not quinic acid inhibited TNF-α and IL-17 levels (p < 0.05). C. antisyphiliticus caused anti-inflammatory effect by inhibiting the activated leukocytes, exudate concentrations, NOx, TNF-α, and IL-17 levels. The compounds vitexin and quinic acid may be responsible for this anti-inflammatory action.     

人参皂甙Rg1对大鼠关节软骨细胞凋亡的拮抗作用

目的 探讨人参皂甙Rg1对软骨细胞凋亡的影响.方法 正常大鼠关节软骨细胞分为三组:A、B组用白细胞介素1β(IL-1β)10 ng/ml作用24 h,建立软骨细胞凋亡模型;B组事先加入10 μg/ml的人参皂甙Rg1保护2h;C组作为空白对照.TUNEL法检测人参皂甙Rg1对软骨细胞凋亡的影响;Western blot检测人参皂甙Rg1对凋亡软骨细胞基质金属蛋白酶13(MMP-13)和基质金属蛋白酶组织抑制剂1(TIMP-1)蛋白表达的影响.结果 A、B和C组细胞凋亡率分别为(37.30±4.05)％、(10.30±1.08)％和(2.20±1.93)％,B组胞凋亡率低于A组(P＜0.05).与A组比较,B组软骨细胞TIMP-1的表达增加,MMP-13的表达降低(P＜0.05).结论 人参皂甙Rg1具有拮抗大鼠关节软骨细胞凋亡的作用.     

西洋参中人参皂苷Rg1人参皂苷Re和人参皂苷Rb1的含量测定

［摘要］目的用快速分离液相色谱法分离测定西洋参中人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1的含量。方法采用ZOBAX SB C18柱（4.6 mm×50 mm,1.8 μm）,流动相：乙腈（A） 0.1%磷酸（B）,梯度洗脱（0～25min,5%A→20%A;～35min,20%A→40%A）;流速：2.0 mL&#8226;min 1,测定波长：203 nm,柱温：35 ℃。结果人参皂苷Rg1、人参皂苷Re和人参皂苷Rb1的线性范围分别为0.12～2.49,0.61～12.15,1.42～28.48 μg,相关系数分别为0.999 8,0.999 6,0.999 9;平均加样回收率（n=6）分别为97.8%,98.1%,98.3%;RSD分别为1.0%,0.8%,0.4%。结论该方法具有快速、准确、重复性好等特点,适合于西洋参的含量测定。     

RP-HPLC法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量

目的：建立反相高效液相色谱法测定三七丹胶囊中人参皂苷Rb1、人参皂苷Rg1和三七皂苷R1的含量测定方法。方法：采用Hibar ODS C18（4.6mm×250mm,5μm）色谱柱,流动相为乙腈-水,梯度洗脱,流速1.0ml·min-1,柱温30℃,检测波长203nm。结果：人参皂苷Rb1、人参皂苷Rg1及三七皂苷R1分别在0.5246～5.2464μg（r=0.9997）、0.6792～6.7920μg（r=0.9998）和0.2542～2.5424μg（r=0.9997）范围内线性关系良好,平均回收率分别为人参皂苷Rb199.12%（RSD=0.61%,n=6）、人参皂苷Rg197.50%（RSD=1.63%,n=6）、三七皂苷R197.43%（RSD=1.04%,n=6）。结论：该方法定量简单、准确、重复性好,可作为三七丹胶囊的质量控制方法。     

高效液相色谱法测定活血止痛胶囊中人参皂苷Rg1含量

目的建立测定活血止痛胶囊中人参皂苷Rg1含量的高效液相色谱（HPLC）法。方法采用Zorbax Extend—C18柱（150mm×4．6mm,5μm）,流动相为乙腈-0．1％磷酸溶液（20：80）,流速为1．0mL／min,检测波长为203nm,柱温为30℃。结果人参皂苷Rg1进样量在1．236～9．888μg（r=0．9991）范围内与峰面积线性关系良好,平均回收率为97．90％,RSD=1．03％（n=6）。结论HPLC法简便可靠、专属性强,可用于活血止痛胶囊的质量检测。