p-Cymene Modulates In Vitro and In Vivo Cytokine Production by Inhibiting MAPK and NF-κB Activation

The present study was designed to investigate the effects of p-cymene on lipopolysaccharide (LPS)-induced inflammatory cytokine production both in vitro and in vivo. The production of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interleukin-10 (IL-10) in LPS-stimulated RAW 264.7 cells and C57BL/6 mice was evaluated by sandwich ELISA. Meanwhile, the mRNA levels of cytokine genes were examined in vitro by semiquantitative RT-PCR. In a further study, we analyzed the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways by western blotting. We found that p-cymene significantly regulated TNF-α, IL-1β, and IL-6 production in LPS-stimulated RAW 264.7 cells. Furthermore, the levels of relative mRNAs were also found to be downregulated. In in vivo trail, p-cymene markedly suppressed the production of TNF-α and IL-1β and increased IL-10 secretion. We also found that p-cymene inhibited LPS-induced activation of extracellular signal receptor-activated kinase 1/2, p38, c-Jun N-terminal kinase, and IκBα. These results suggest that p-cymene may have a potential anti-inflammatory action on cytokine production by blocking NF-κB and MAPK signaling pathways.     

计算机辅助设计纳米羟基磷灰石／聚氨酯复合骨植入物的个性化快速制造

采用原位复合法,以纳米羟基磷灰石为增强体,以聚氨酯预聚体为基体,制备了快速成型所需的HA /PU 基质复合材料,采集个体股骨近端骨缺损部位的 CT 影像数据,利用计算机辅助医学影像 Mimics 软件进行三维重建,直接得到股骨近端的三维模型,再将三维模型信息转换成快速成型的“STL”数据文件,以此直接驱动三维打印机制造出 ABS 实体模型,然后经硅胶模具和浇注成型手段,将 HA /PU 复合预聚体基质浇注到硅胶模具中,进行固化成型,得到符合个性化需求的 HA /PU 股骨近端植入体。为克服批量生产植入体型号、尺寸与个体需求不匹配的弊端,制造高附加值人工骨产品提供实验基础和技术支持。     

Inhibition of lung inflammatory responses by bornyl acetate is correlated with regulation of myeloperoxidase activity

Background

Bornyl acetate is a bicyclic monoterpene present in numerous conifer oils. In this study, we aimed at clarifying the potential anti-inflammatory function and mechanism of bornyl acetate by using lipopolysaccharide (LPS)-induced acute lung injury murine model and RAW 264.7 cells.

Materials and methods

RAW 264.7 cells were pretreated with bornyl acetate 1 h before LPS stimulation and cell-free super supernatants were collected to measure cytokine concentrations. To induce acute lung injury, BALB/c mice were injected intranasally with LPS and treated with bornyl acetate 1 h before LPS stimulation. Seven hours after administration, the bronchoalveolar lavage fluid (BALF) was collected for measuring the cell count and cytokine production. We collected lungs for assaying wet-to-dry weight ratio, myeloperoxidase activity, and histologic changes. The extent of phosphorylation of mitogen-activated protein kinases and nuclear factor κB was detected by Western blot.

Results

Our results showed that bornyl acetate downregulated the levels of proinflammatory cytokines in vitro and in vivo; reduced the number of total cells, neutrophils, and macrophages in BALF; attenuated the histologic alterations in the lung; decreased the wet-to-dry weight ratio in BALF; and suppressed NF-kappa-B inhibitor alpha, extracellular regulated protein kinases, c-JunN-terminal kinase, p38 mitogen-activated protein kinase activation.

Conclusions

These findings suggested that bornyl acetate may be developed as a preventive agent for lung inflammatory diseases.     

In vitro and in vivo protection provided by pinocembrin against lipopolysaccharide-induced inflammatory responses

Pinocembrin or 5, 7-dihydroxyflavanone is a flavanone, a type of flavonoid. In the present study, we first assessed the anti-inflammatory effects of pinocembrin in RAW macrophage cells; and based on these effects, we investigated the therapeutic effects of pinocembrin in murine model of endotoxin-induced acute lung injury. We found that in vitro pretreatment with pinocembrin remarkably regulated the production of TNF-α, IL-1β, IL-6 and IL-10 via inhibiting the phosphorylation of IκBα, ERK1/2, JNK and p38MAPK. In the mouse model of LPS-induced acute lung injury, pinocembrin (20 or 50 mg/kg, i.p.) attenuated the development of pulmonary edema, histological severities, as well as neutrophil, lymphocyte and macrophage infiltration, which were increased by LPS administration. Additionally, TNF-α, IL-1β and IL-6 concentrations decreased significantly while the concentration of IL-10 was significantly increased after pinocembrin pretreatment. Our results also showed that pinocembrin attenuated LPS-induced lung injury through suppression of IκBα, JNK and p38MAPK activation. These findings suggest that pinocembrin may represent a novel candidate for the modulation of inflammatory responses.     

Traditional medicine alpinetin inhibits the inflammatory response in Raw 264.7 cells and mouse models

Alpinetin, one of the main constituents of the seeds of Alpinia katsumadai Hayata, belonging to flavonoids, has been known to exhibit antibacterial, anti-inflammatory and other important therapeutic activities. The purpose of this study was to investigate the protection of alpinetin on inflammation in Lipopolysaccharide (LPS) stimulated Raw 264.7 cells and LPS induced vivo lung injury model. The effects of alpinetin on pro-inflammatory cytokines and signaling pathways were analyzed by enzyme-linked immunosorbent assay and Western blot. The results showed that alpinetin markedly inhibited the LPS- induced TNF-α, IL-6 and IL-1β production both in vitro and vivo. Furthermore, alpinetin blocked the phosphorylation of IκBα protein, p65, p38 and extracellular signal-regulated kinase (ERK) in LPS stimulated RAW 264.7 cells. From in vivo study, it was also observed that alpinetin attenuated lung histopathologic changes in mouse models. These results suggest that alpinetin potentially decreases the inflammation in vitro and vivo, and might be a therapeutic agent against inflammatory diseases.     

Imperatorin Attenuates LPS-Induced Inflammation by Suppressing NF-κB and MAPKs Activation in RAW 264.7 Macrophages

Imperatorin is a type of coumarin compound with antibacterial and antiviral activities. In the present study, we examined the anti-inflammatory effects of imperatorin in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by investigating its impact on the production and expression of cytokines and the major signal-transduction pathways. We found that imperatorin downregulated LPS-induced levels of TNF-??, IL-1??, and IL-6 in RAW 264.7 macrophages in a concentration-dependent manner, and it significantly inhibited expression of TNF-?? and IL-6 (P?<?0.05 or P?<?0.01). The phosphorylation of mitogen-activated protein kinases and nuclear factor-kappaB (NF-??B) p65 protein were analyzed by western blotting. In RAW 264.7 macrophages treated with 1?mg/L of LPS, imperatorin significantly inhibited p38 and Jun N-terminal kinase phosphorylation protein expression. However, there was no significant change in p-ERK. Furthermore, imperatorin also inhibited NF-??B translocation into the nucleus through blockage of I??B?? phosphorylation and degradation.     

人肺腺癌多药抗药细胞系LC-3/CDDP细胞周期及DNA含量变化

应用流式细胞仪测定人肺腺癌多药抗药细胞系LC－3／CDDP之细胞周期、DNA指数，用3H-胸腺嘧啶核苷掺入法测定DNA合成动态，结果显示，LC－3／CDDP细胞系G2＋M峰较亲代细胞明显增高，G期细胞比例降低，S期、G2＋M期比例升高，DNA指数增加，DNA合成速率较亲代细胞显著增快。提示人肺腺癌多药抗药细胞系LC－3／CDDP对DNA损伤的修复功能增强。     

p-Synephrine suppresses lipopolysaccharide-induced acute lung injury by inhibition of the NF-κB signaling pathway

Objective

We investigated whether p-synephrine exerts potent anti-inflammatory effects against acute lung injury (ALI) induced by lipopolysaccharide (LPS) in vivo, and we further investigated the inhibitory mechanism of p-synephrine in LPS-induced ALI.

Methods

Lipopolysaccharide (0.5 mg/kg) was instilled intranasally in phosphate-buffered saline to induce acute lung injury, and 6, 24, and 48 h after LPS was given, bronchoalveolar lavage fluid was obtained to measure pro-inflammatory mediator. We also evaluated the effects of p-synephrine on LPS-induced the severity of pulmonary injury. The phosphorylation of nuclear factor-κB (NF-κB) p65 protein was analyzed by Western blotting.

Results

Our data showed that p-synephrine significantly reduced the amount of inflammatory cells, the lung wet-to-dry weight (W/D) ratio, reactive oxygen species, myeloperoxidase activity and enhanced superoxide dismutase (SOD) in mice with LPS-induced ALI. Tumor necrosis factor α and interleukin (IL)-6 concentrations decreased significantly while the concentration of IL-10 was significantly increased after p-synephrine pretreatment. In addition, p-synephrine suppressed not only the phosphorylation of NF-κB but also the degradation of its inhibitor (IκBα).

Conclusions

These results suggested that the inhibition of NF-κB activation and the regulation of SOD are involved in the mechanism of p-synephrine’s protection against ALI.     

Paeonol suppresses lipopolysaccharide‐induced inflammatory cytokines in macrophage cells and protects mice from lethal endotoxin shock

Paeonol (2′‐hydroxy‐4′‐methoxyacetophenone) is the main phenolic compound of the radix of Paeonia suffruticosa which has been used as traditional Chinese medicine. In this study, we primarily investigated the anti‐inflammatory effects and the underlying mechanisms of paeonol in RAW macrophage cells; and based on these effects, we assessed the protective effects of paeonol on lipopolysaccharide‐induced endotoxemia in mice. The in vitro study showed that paeonol regulated the production of TNF‐α, IL‐1β, IL‐6, and IL‐10 via inactivation of IκBα, ERK1/2, JNK, and p38 MAPK. In mouse model of lipopolysaccharide‐induced endotoxemia, pro‐ and anti‐inflammatory cytokines are significantly regulated, and thus the survival rates of lipolysaccharide‐challenged mice are improved by paeonol (150, 200, or 250 mg/kg). Therefore, paeonol has a beneficial activity against lipopolysaccharide‐induced inflammation in RAW 264.7 cell and mouse models.