Down‐regulatory effect of usnic acid on nuclear factor‐κB‐dependent tumor necrosis factor‐α and inducible nitric oxide synthase expression in lipopolysaccharide‐stimulated macrophages RAW 264.7

The purpose of this study was to investigate the molecular mechanisms that are responsible for the antiinflammatory effect of usnic acid (UA). UA is one of the most common and abundant lichen metabolites. The present study examined the effects of UA on the tumor necrosis factor‐α (TNF‐α) and nitric oxide (NO) production induced by lipopolysaccharide (LPS) in RAW264.7 macrophages and the underlying molecular mechanisms. UA decreased the TNF‐α level in LPS‐stimulated RAW264.7 macrophages in dose‐dependent manner, the IC₅₀ value was 12.8 µM. RT‐PCR analysis indicated that it inhibited TNF‐α mRNA expression. Furthermore, it inhibited NO production in LPS‐activated RAW264.7 macrophages, the IC₅₀ value was 4.7 µM. Western blot analysis showed that UA attenuated LPS‐induced synthesis of iNOS protein and nuclear translocation of NF‐κB p65 in the macrophages, in parallel. UA also inhibited LPS‐mediated I‐κBα degradation. Taken together, this suggests that UA has an antiinflammatory effect by inhibiting TNF‐α and iNOS expression, possibly through suppression of nuclear translocation of NF‐κB p65 and I‐κBα degradation. Copyright © 2008 John Wiley & Sons, Ltd.     

Antiinflammatory Constituents from Eclipta prostrata using RAW264.7 Macrophage Cells

The whole plant extract of Eclipta prostrata and its isolated compounds were tested for their antiinflammatory effects against lipopolysaccharide (LPS)‐induced nitric oxide (NO), prostaglandin E₂ (PGE₂) and tumor necrosis factor‐alpha (TNF‐α) release in RAW264.7 cells, as well as for the antiinflammatory mechanism of the active compound on mRNA expression. Among the isolated compounds, orobol (5) exhibited the highest activity against NO release with an IC₅₀ value of 4.6 μm , followed by compounds 1, 2 and 4 with IC₅₀ values of 12.7, 14.9 and 19.1 μm , respectively. The IC₅₀ value of compound 5 against PGE₂ release was found to be 49.6 μm , whereas it was inactive towards TNF‐α (IC₅₀ > 100 μm ). The mechanism of orobol (5) was found to down‐regulate iNOS and COX‐2 mRNA expression in a concentration‐dependent manner. The present study may support the traditional use of Eclipta prostrata for the treatment of inflammatory‐related diseases. Copyright © 2011 John Wiley & Sons, Ltd.     

Inhibition of Inducible Nitric Oxide Synthase and Cyclooxygenase‐2 Expression by Phenolic Compounds from Roots of Rhododendron mucronulatum

The roots of Rhododendron mucronulatum Turzaninov have been used in Oriental traditional medicine for the treatment of dysuria, fever, increase of digestive activity and tonics in China and Korea. Activity guided isolation of the roots of Rhododendron mucronulatum Turzaninov has led to the isolation of three flavonoids, one flavan 3‐ol and one proanthocyanidin. Chemical investigation of the 80% Me₂CO extract from the roots of Rhododendron mucronulatum led to the isolation and identification of five compounds: taxifolin (1), taxifolin 3‐O‐β‐d ‐glucopyranoside (2), quercetin 3‐O‐α‐l ‐arabinofuranoside (3), (‐)‐epicatechin (4), procyanidin B‐3 (5). To investigate the antioxidative and antiinflammatory effects of these compounds, their 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging activities and the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) in LPS‐stimulated HaCaT cells were also quantified by western blotting and their end products, nitric oxide (NO) and prostaglandin E₂ (PGE₂), respectively. Compounds (1–5) showed potent DPPH radical scavenging compared with positive controls (l ‐ascorbic acid). Also, compounds 1 and 2 dose‐dependently inhibited the expressions of inflammatory mediators, NO and PGE₂, suggesting they are promising candidates as antiinflammatory agents. Copyright © 2011 John Wiley & Sons, Ltd.     

The Antiinflammatory Effect of an Extract of Tripterygium wilfordii Hook F on Adjuvant-induced Paw Oedema in Rats and Inflammatory Mediators Release

Tripterygium wilfordii Hook F (TWH), commonly known as the Thunder-God-Vine, is a popular remedy for rheumatism in southern China. In this study, we investigated the effect of T2 (an extract from TWH) on adjuvant-induced paw oedema in rats and inflammatory mediators. The results showed that an intraperitoneal injection of T2 could significantly decrease the adjuvant-induced paw oedema of rats. During in vitro study, lipopolysaccharide (LPS)-stimulated human mononuclear prostaglandin E₂ (PGE₂) in the culture supernatant was significantly inhibited by T2 at a concentration of 2 μg/mL compared with the control group without T2 treatment (22352±4725 vs 43433±9014 pg/mL, p <0.05). When rat neutrophils were treated with 3 μg/mL of T2, the superoxide production was significantly lower than that of the group without T2 treatment (0.72±0.07 vs 1.37±0.04 nmol, p <0.05). T2 failed to suppress the β-glucuronidase release from rat neutrophils but when the concentration was 30 μg/mL, it expressed a significant inhibition of lysozyme release (20.7±2.9 vs 38.2±4.0 in the control, p <0.05). The findings suggest this Chinese herb, like non-steroid antiinflammatory drugs, expresses a potential antiinflammatory effect through its inhibition of PGE₂ and superoxide function and lysozyme release. © 1997 by John Wiley & Sons, Ltd.     

Antiinflammatory,antitumour and antifertility effects in rodents of two nor-cucurbitacin glucosides from Wilbrandia species

The purified fraction (PF) of the rhizome of Wilbrandia sp. (Cucurbitaceae) that contained two nor-cucurbitacin glucosides (WG₁ and WG₂) demonstrated potent antiinflammatory, antitumour and antifertility effects in rats and mice at oral doses ranging from 50 to 200 mg/kg. In rat, it significantly inhibited carrageenan-induced hind paw oedema and granulomatous lesion, while in mice the increased vascular permeability induced by acetic acid was markedly reduced. The PF exhibited an antitumour effect in rats bearing Walker 256 carcinosarcoma and also demonstrated cytotoxic action against KB cells in vitro. In regularly cycling mice, PF treatment suppressed the incidence of the oestrus phase of the reproductive cycle suggesting a possible antiovulatory effect. The rats that received PF from days 1 to 7 of pregnancy showed either a total absence or a reduced number of implantations in utero. The PF failed to exhibit abortifacient, oestrogenic or antioestrogenic actions. The results indicate that the nor-cucurbitacin glucosides of Wilbrandia sp. possess a novel spectrum of pharmacological activity different from that of cucurbitacins described earlier from other plants of this family.     

Optimization of Aqueous Extraction and Biological Activity of Harpagophytum procumbens Root on Ex Vivo Rat Colon Inflammatory Model

Harpagophytum procumbens has a long story of use for the treatment of inflammatory diseases. Considering both the antiinflammatory effects of H. procumbens in multiple tissues and the stability of harpagoside in artificial intestinal fluid, the aim of the present study was to explore the possible protective role of a microwave‐assisted aqueous Harpagophytum extract (1–1000 μg/mL) on mouse myoblast C2C12 and human colorectal adenocarcinoma HCT116 cell lines, and isolated rat colon specimens challenged with lipopolysaccharide (LPS), a validated ex vivo model of acute ulcerative colitis. In this context, we evaluated the effects on C2C12 and HCT116 viability, and on LPS‐induced production of serotonin (5‐HT), tumor necrosis factor (TNF)‐α, prostaglandin (PG)E₂ and 8‐iso‐prostaglandin (8‐iso‐PG)F_2α. Harpagophytum extract was well tolerated by C2C12 cells, while reduced HCT116 colon cancer cell viability. On the other hand, Harpagophytum extract reduced H₂O₂‐induced (1 mM) reactive oxygen species (ROS) production, in both cell lines, and inhibited LPS‐induced colon production of PGE₂, 8‐iso‐PGF_2α, 5‐HT and TNFα. Concluding, we demonstrated the efficacy of a microwave‐assisted Harpagophytum aqueous extract in modulating the inflammatory, oxidative stress and immune response in an experimental model of inflammatory bowel diseases (IBD), thus suggesting a rational use of Harpagophytum in the management and prevention of ulcerative colitis in humans. Copyright © 2017 John Wiley & Sons, Ltd.     

Inhibition of release of inflammatory mediators in rat peritoneal mast cells and murine macrophages by a chinese herbal medicine formula (RCM‐102)

RCM‐102 is a Chinese herbal medicine formulation derived from a formula which was shown to be effective in treating seasonal allergic rhinitis (SAR) in a randomized placebo‐controlled trial. The aim of this study was to investigate the in vitro effect of RCM‐102 on the formation of inflammatory mediators, histamine, prostaglandin and nitric oxide, which are known to be involved in the pathophysiology of SAR. The effect of RCM‐102 on histamine release was tested in compound 48/80‐stimulated rat peritoneal mast cells. The effects of RCM‐102 on the release of NO and prostaglandins (PGE₂) and the expression of inducible NO synthase (iNOS) and COX‐2 were studied in lipopolysaccharide (LPS)‐stimulated RAW 264.7 cells. In rat peritoneal mast cells, RCM‐102 significantly reduced the compound 48/80‐induced histamine release. It also significantly reduced NO and PGE₂ production as well as the expression of COX‐2 and iNOS in RAW 264.7 cells. These findings indicate that RCM‐102 inhibits the formation of several allergic/inflammatory mediators and thus may be used for treating related conditions such as SAR. The actions of RCM‐102 are likely to be contributed by the synergistic actions of individual herbal ingredients. Copyright © 2009 John Wiley & Sons, Ltd.     

Praeruptorin a inhibits lipopolysaccharide‐induced inflammatory response in murine macrophages through inhibition of NF‐κB pathway activation

Praeruptorin A (PA) is a pyranocoumarin compound isolated from the dried root of Peucedanum praeruptorum Dunn (Umbelliferae). However, the antiinflammatory effect of PA has not been reported. The present study investigated the antiinflammatory effect of PA in lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophage cells. PA significantly inhibited the LPS‐induced production of nitric oxide (NO), interleukin‐1β (IL‐1β) and tumor necrosis factor‐α (TNF‐α). The mRNA and protein expressions of inducible nitric oxide synthase (iNOS), IL‐1β and TNF‐α were also suppressed by this compound. Further study showed that PA decreased the cytoplasmic loss of inhibitor κB‐α (IκB‐α) protein and inhibited the translocation of NF‐κB from cytoplasm to nucleus. Taken together, the results suggest that PA may exert antiinflammatory effects in vitro in LPS‐stimulated RAW 264.7 macrophages through inhibition of NF‐κB signal pathway activation. Copyright © 2010 John Wiley & Sons, Ltd.     

Anti‐Arthritic and Antiinflammatory Effects of the Traditional Uighur Formula Kursi Caper In Vivo

Kursi Caper (KC) is a Uighur medicine based on caper which is widely used to treat arthritis and rheumatism, and preliminary studies in our laboratory showed that this traditional formula may possess potent antiinflammatory effects. This study confirms the antiinflammatory effect of KC in the adjuvant induced arthritis (AIA) model, the carrageenan and cotton‐pellet induced granuloma rat models, and further investigates in vivo the mechanism of action by measuring relevant indicators of anti‐arthritic activity. KC showed significant and dose‐dependent anti‐arthritic and antiinflammatory effects, demonstrated by reduced paw edema and arthritic scores in all animal models. Histopathological examination showed that KC reduced levels of synovial inflammatory factors in AIA rats. The overproduction of TNF‐α and IL‐1β was attenuated, and CAT, MDA and SOD levels were restored to normal in KC‐treated rats. KC also significantly reduced LPS‐induced proliferation of B lymphocytes and ConA induced proliferation of T lymphocytes in a dose‐dependent manner. Flow cytometry showed that the high dose KC‐treated group had a significantly decreased frequency of Th17 cells. This study indicates that KC can significantly attenuate arthritis and inflammation in rats by decreasing the levels of inflammatory cytokines, regulating oxidative stress, reducing lymphocyte proliferation and decreasing Th17. This supports the traditional use of KC as a potential modern therapeutic agent for the treatment of arthritis and related conditions. Copyright © 2015 John Wiley & Sons, Ltd.     

Effect of an extract of Andrographis paniculata leaves on inflammatory and allergic mediators in vitro

Aim of study

Andrographis paniculata has been known to possess widespread traditional application in the treatment of allergy and inflammatory diseases. In the current study, we sought to examine the effects of an extract of Andrographis paniculata leaves on inhibition of lipopolysaccharide (LPS) induced [nitric oxide (NO), prostaglandin E₂ (PGE₂), interleukin-1beta (IL-1 beta), and interleukin-6 (IL-6)] and calcimycin (A23187) induced [leukotriene B₄ (LTB₄), thromboxane B₂ (TXB₂) and histamine] mediators in diverse cell based models.

Materials and methods

Effect of an extract of Andrographis paniculata leaves (AP) was studied on inhibition of LPS induced NO, PGE₂, IL-1 beta and IL-6 in J774A.1 murine macrophages; A23187 induced LTB₄ and TXB₂ in HL-60 promyelocytic leukemic cells and histamine in RBL-2H3 rat basophilic leukemia cells.

Results and conclusion

AP illustrated significant alleviation of pro-inflammatory, inflammatory, and allergic mediators. However, no inhibition was observed against histamine release. This outcome has been summed up to deduce that AP is fairly potent in attenuating the inflammation by inhibiting pro-inflammatory (NO, IL-1 beta and IL-6), inflammatory (PGE₂ and TXB₂) and allergic (LTB₄) mediators.     

黄连解毒汤的抗炎作用机理研究

目的 :研究黄连解毒汤的抗炎机理。方法 :采用小鼠背部气囊内注射角叉菜胶的方法诱导急性气囊滑膜炎症 ;脂多糖腹腔注射造成内毒素血症模型 ;噻唑蓝比色法测定细胞增殖 ;Griess法检测上清中NO^-₂ 的浓度 ;胸腺细胞增殖测定IL 1的活性。结果 :黄连解毒汤能抑制角叉菜胶所致小鼠气囊内白细胞的游出 ,减少PGE2 的生成。在体外实验中 ,黄连解毒汤能显著抑制ConA所致的内毒素血症小鼠脾淋巴细胞的增殖 ,但对正常小鼠淋巴细胞的增殖无影响 ,且不影响正常及内毒素血症小鼠的脾细胞生成IL₂。黄连解毒汤还可抑制脂多糖诱导小鼠腹腔巨噬细胞生成IL-1和NO。结论 :以上结果提示黄连解毒汤的抗炎作用主要与抑制IL-1 ,NO ,PGE₂ 等炎症因子生成有关。     

Colon Cancer Chemoprevention by Sage Tea Drinking: Decreased DNA Damage and Cell Proliferation

Salvia officinalis and some of its isolated compounds have been found to be preventive of DNA damage and increased proliferation in vitro in colon cells. In the present study, we used the azoxymethane model to test effects of S. officinalis on colon cancer prevention in vivo. The results showed that sage treatment reduced the number of ACF formed only if administered before azoxymethane injection, demonstrating that sage tea drinking has a chemopreventive effect on colorectal cancer. A decrease in the proliferation marker Ki67 and in H₂O₂‐induced and azoxymethane‐induced DNA damage to colonocytes and lymphocytes were found with sage treatment. This confirms in vivo the chemopreventive effects of S. officinalis. Taken together, our results show that sage treatment prevented initiation phases of colon carcinogenesis, an effect due, at least in part, to DNA protection, and reduced proliferation rates of colon epithelial cell that prevent mutations and their fixation through cell replication. These chemopreventive effects of S. officinalis on colon cancer add to the many health benefits attributed to sage and encourage its consumption. Copyright © 2015 John Wiley & Sons, Ltd.     

千金苇茎汤促进撤血清舌鳞癌细胞凋亡机制的研究

目的: 探讨千金苇茎汤辅助治疗舌鳞癌的分子机制。方法: 体外培养SAS和TCA-8113舌鳞癌细胞,撤除血清模拟体内抗血管生成药物治疗环境,千金苇茎汤3个部位作用撤血清舌鳞癌细胞72 h,MTT法检测细胞增殖活性,流式细胞术检测细胞周期分布和凋亡,ELISA检测细胞培养上清前列腺素E₂(PGE₂)含量。结果: 与撤血清对照相比,千金苇茎汤07部位能够降低撤血清舌鳞癌细胞的增殖活性(生长抑制率_max34.36%),促进SAS和TCA-8113细胞凋亡率升高(P<0.05),抑制细胞分泌PGE₂(P<0.05),且具有剂量和时间依赖性。结论: 千金苇茎汤07部位促进撤血清舌鳞癌细胞凋亡的机制与抑制PGE₂合成相关,为深入研究千金苇茎汤辅助治疗恶性肿瘤提供了新的思路和方法。     

蜂胶的外用抗炎作用实验研究

目的:观察蜂胶的外用抗炎作用.方法:昆明种小鼠60只,随机分为模型对照组(3.0 g·kg-1吐温80)、阳性药布洛芬乳膏组(4.0 g·kg-1),蜂胶高、中、低剂量组(0.72,0.24,0.08 g·kg-1),每组12只采用二甲苯致小鼠耳廓肿胀模型,检测小鼠血清中前列腺素E2(PGE2),一氧化氮(N0)含量,观察蜂胶对二甲苯致小鼠耳廓肿胀炎症模型的影响;Wistar大鼠60只,分组同上,模型对照组(3.0 g·kg-1吐温80),布洛芬乳膏组(3.0g·kg-1),蜂胶高、中、低剂量组(0.20,0.10,0.05 g·kg-1),采用角叉菜胶致大鼠足跖肿胀模型,检测大鼠炎症组织中PGE2含量,观察蜂胶对角叉菜胶致大鼠足跖肿胀炎症模型的影响.结果:蜂胶能明显抑制二甲苯导致的小鼠耳肿胀(P＜0.05);能明显减轻角叉菜胶导致的大鼠足肿胀(P＜0.05);能显著降低小鼠血清中PGE2,NO含量(P＜0.01);能显著降低大鼠炎症组织中PGE2含量(P＜0.01).结论:蜂胶外用具有明显的抗炎作用.     

双氯芬酸选择性抑制表达环氧合酶-2的肝癌细胞的增殖

 目的检测体外培养的人肝细胞癌HepG2,Hep3B细胞和正常肝细胞系QSG7701细胞内环氧酶-2(cyclooxygenase-2,COX-2)的表达,观察环氧酶抑制剂双氯芬酸(diclofenac)对上述细胞增殖的作用,并检测前列腺素E₂(PGE₂)对双氯芬酸作用的影响,以探讨双氯芬酸抗增殖作用与细胞内COX-2的关系。方法采用半定量逆转录聚合酶链反应(RT-PCR)检测COX-2 mRNA的表达水平,免疫印迹技术检测细胞内蛋白水平,cellcounting kit-8(CCK-8)细胞计数法测定细胞增殖活性。结果肝细胞癌HepG2和Hep3B细胞均高表达COX-2 mRNA和COX-2蛋白,正常肝细胞系QSG7701微弱表达COX-2。10～200μmol·L^-1的双氯芬酸浓度依赖性地抑制HepG2和Hep3B细胞的增殖(P<0.01),作用72h的半数抑制浓度(IC₅₀)分别为76.41和35.21μmol·L^-1。双氯芬酸对QSG7701的抑制作用较弱,IC₅₀值为170.23μmol·L^-1。20μmol·L^-1 PGE2与50μmol·L^-1双氯芬酸共同孵育能显著削弱后者抗HepG2和Hep3B细胞增殖的作用(P<0.01)。结论双氯芬酸通过抑制COX-2活性和PGE₂的生成,特异性地抑制表达COX-2的肝癌细胞的增殖。     

新痹痛灵抗炎作用及其机制

目的:研究新痹痛灵(New Bitongling,NBTL)的抗炎作用,揭示其抗炎作用及机制。方法:体内实验取30只SD雄性大鼠,随机分成模型组、阳性药布洛芬缓释胶囊组(0.054 g·kg-1)以及新痹痛灵高剂量组(11 g·kg-1,简称NBTLH)、低剂量组(2.75 g·kg-1,简称NBTLL);连续ig给药5 d,1次/d;末次给药1 h后,复制组胺致大鼠毛细血管通透性增加模型、采用尾静脉注射伊文斯蓝后提取皮肤中染料、测定提取液590 nm的吸光度反映毛细血管通透性;复制角叉菜胶致大鼠足跖肿胀模型、采用容积测量法观察各组大鼠足跖肿胀度;体外实验以脂多糖(LPS)诱导的RAW 264.7细胞分泌炎性因子复制炎症模型,酶联免疫吸附试验(ELISA)法测定各组细胞上清液中前列腺素E2(PGE2)含量。结果:与模型组比较,布洛芬缓释胶囊组,NBTLH,NBTLL组伊文斯蓝含量明显降低(P0.01);表明均能抑制大鼠毛细血管通透性增加;与模型组相比,在相同时间点,NBTLH组大鼠足跖肿胀度均降低(各时间点P0.05),NBTLL在各个时间点都有抑制趋势,但无显著性差异;与正常组比较,模型组的PGE2表达水平升高(P0.05);与模型组比较,NBTLH,NBTLL组PGE2表达水平显著降低(P0.05,P0.01)。结论:NBTL对炎症有明显抑制作用,其抗炎作用可能与抑制炎症组织PGE2的合成或分泌有关。