铁观音茶提取物对脂多糖诱导RAW264.7细胞炎症反应的抑制作用及机制

研究铁观音茶提取物对脂多糖(LPS)诱导的RAW264.7细胞炎症反应的抑制作用及机制。用脂多糖作用于RAW264.7细胞,建立炎症模型,并用吲哚美辛和不同浓度铁观音提取物处理,检测NO和IL-6的分泌情况,qPCR检测一氧化氮合酶(iNOS)、环氧合酶2(COX-2)、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)、白细胞介素6(IL-6)mRNA相对表达,Western Blot检测炎症相关蛋白激酶(IKKβ),核转录因子κB抑制因子(IκB)、核转录因子κB p65(NF κB p65)及其磷酸化产物的相对表达。结果显示,铁观音茶提取物能显著抑制炎症介质NO分泌和IL-6蛋白表达量(p<0.05),抑制炎症相关基因iNOS、COX-2、TNF-α和MCP-1等表达,并极显著抑制NF-κB信号通路相关蛋白IKKβ、IkB和p65的磷酸化(p<0.01)。以上结果表明,铁观音茶提取物可明显抑制LPS诱导的RAW264.7细胞炎症反应,其机制可能与抑制NF-κB信号通路激活有关。     

Anti-inflammatory effect of phenethyl isothiocyanate, an active ingredient of Raphanus sativus Linne

Phenethyl isothiocyanate (PEITC) is an active ingredient of Raphanus sativus Linne (Cruciferae). However, regulatory mechanism of PEITC involved in caspase-1 signalling has not been fully elucidated in mast cells. First, PEITC inhibited the production of IL-6 through the inhibition of caspase-1/receptor-interacting protein 2, followed by regulation of NF-κB/IκBα pathway or p38 and extracellular signal-regulated kinase mitogen-activated protein kinases. Second, PEITC inhibited the IL-1β production through the inhibition of caspase-1 proteolytic activity. Overall, these results provide a proof that PEITC can inhibit the inflammatory reactions by two distinct pathways in mast cells and open new perspectives to pharmacologically manipulate the expression and production of IL-6 and IL-1β by molecules acting on the caspase-1 pathway.     

Anti-inflammatory activity of ethanolic extract of Sargassum sagamianum in RAW 264.7 cells

Aim of the study is to evaluate the antiinflammatory effects of ethanolic extract of the marine brown alga Sargassum sagamianum collected from Yeonhwari coast of Korea. Ethanolic extract of S. sagamianum (SA-E extract) inhibited expression of nitric oxide (NO) and cytokines (IL-6, IL-1β, and TNF-α) as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. In addition, the expression of nuclear factor (NF)-κB p65 was suppressed by SA-E extract. Furthermore, the rate of formation of edema in the mouse ear was reduced by 46% at the highest dose tested (250 mg/kg) compared to that in the control. This study suggests that SA-E extract exerts potent inhibitory effects on LPS-induced expression of inflammatory mediators such as NO, iNOS, COX-2, and cytokines in macrophages through suppression of the NF-κB p65 pathway. SA-E extract might have potential clinical applications as an anti-inflammatory agent.     

Extrusion process enhances the anti-inflammatory effect of Acanthopanax senticosus leaves

The anti-inflammatory effects of Acanthopanax senticosus leaves (ASL) and the effects of extrusion were evaluated. ASL exhibited in vitro and in vivo antiinflammatory activities in experimental systems. Extrusion increased the effect of ASL. ASL reduced C48/80-induced histamine release from HMC-1 cells as well as in vivo model, suggesting that ASL induces mast cell stabilization and has an anti-histamine activity. The effects of ASL and extruded ASL (ASLE) on pro-inflammatory cytokine production were evaluated. ASL treatment reduced MCP- 1, TNF-α, and IL-1β mRNA expressions and decreased their protein levels in HMC-1 cells. Decreases in serum NO, MDA, and TNF-α levels were observed in acute inflammatory rats, and extrusion increased the effects of ASL in a dose-dependent manner. These data support pharmacological basis of ASL and the effect of extrusion for future treatment of inflammation.     

Suppression of thymic stromal lymphopoietin production by rutin in mast cells

Thymic stromal lymphopoietin (TSLP) is a key player in allergic diseases such as asthma and atopic dermatitis. Rutin (RU), a non-nutritive component of many foods, possesses anti-inflammatory, hepatoprotective, and anti-tumour effects. We investigated how RU inhibits the production of TSLP in human mast cell line (HMC-1) cells. RU inhibited the production and mRNA expression of TSLP in HMC-1 cells. The maximal inhibition rate of TSLP production by RU (50 μM) was 56.25 ± 2.81%. Nuclear factor-κB luciferase activity induced by phorbol myristate acetate plus A23187 was inhibited by RU. In the activated HMC-1 cells, the activation of caspase-1 was increased, whereas the activation of caspase-1 was decreased by pre-treatment with RU. Finally, RU inhibited the numbers of TSLP-positive mast cells in lesions of PMA-induced ear oedema model. These results suggest that RU would be helpful for the treatment of inflammatory and atopic diseases through the inhibition of TSLP.     

榆干离褶伞溶栓酶对脂多糖诱导的血管内皮细胞损伤的保护作用

目的：探讨榆干离褶伞溶栓酶对脂多糖（lipopolysaccharide,LPS）诱导的血管内皮细胞炎性损伤的保护作用。方法：以LPS诱导人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVEC）炎性损伤。将HUVEC分为空白对照组、模型组和榆干离褶伞溶栓酶（Lyophyllum ulmarium fibrinolytic enzyme,LUFE）低、中、高剂量组。采用噻唑蓝法测定HUVEC存活率,通过酶联免疫吸附测试法检测细胞上清液乳酸脱氢酶（lactate dehydrogenase,LDH）、肿瘤坏死因子α（tumor necrosis factor α,TNF-α）、白介素6（interleukin 6,IL-6）、E-选择素和单核细胞趋化因子1（monocyte chemoattractant protein 1,MCP-1）水平。流式细胞术检测细胞间黏附分子1（intercellular cell adhesion molecule 1,ICAM-1）表达水平,采用Hoechst染色法观察HUVEC与人急性单核细胞白血病细胞系（human acute monocytic leukemia cell line-1,THP-1）的黏附作用。用蛋白印迹实验检测HUVEC的Toll样受体4（toll-like receptor 4,TLR4）、丝裂原活化蛋白激酶（mitogen-activated protein kinase,MAPK）以及核因子-κB（nuclear factor κB,NF-κB）通路中主要蛋白（髓样分化因子88（myeloid differentiation factor 88,MyD88）、转化生长因子β激活激酶1（transforming growth factor β activated kinase 1,TAK1）、磷酸化TAK1（phosphorylated TAK1,p-TAK1））的表达和活化情况。结果：LUFE能够抑制LPS所诱导的HUVEC培养上清液LDH、TNF-α、IL-6、E-选择素和MCP-1水平的升高,降低细胞ICAM-1表达水平并减弱HUVEC与THP-1的黏附作用。与模型组比较,LUFE各剂量组TLR4、MyD88、p-TAK1/TAK1、磷酸化c-Jun氨基末端激酶（phosphorylated c-Jun N-terminal kinase,p-JNK）/JNK、p-p38/p38、p-NF-κB/NF-κB水平显著降低（P＜0.05）。结论：LUFE对血管内皮细胞的炎性损伤具有保护作用,其作用机制可能是通过抑制TLR4/MyD88/TAK1/NF-κB信号通路及MAPK通路,进而降低炎症因子水平,从而保护血管内皮细胞。     

黑米花色苷矢车菊素-3-O-β-葡萄糖苷对脂多糖诱导的人THP-1单核细胞炎症损伤的保护作用

单核细胞介导的炎症反应在动脉粥样硬化发生和发展过程中起关键作用。花色苷是一种具有多种生物学活性的多酚类黄酮化合物,富含于各种深色的蔬菜、水果及谷类中,其中矢车菊素-3-O-β-葡萄糖苷（cyanidin-3-O-β-glucoside,Cy-3-g）是花色苷中重要的单体,本研究旨在探讨黑米来源的Cy-3-g对脂多糖（lipopolysaccharide,LPS）诱导的人单核白血病细胞（Tohoku hospital pediatrics-1,THP-1）炎症损伤的作用及可能的分子机制。采用乙醇-盐酸溶液浸提、大孔树脂吸附洗脱等步骤得到黑米花色苷粗提物,然后用中压液相层析联合紫外检测提取得到纯化的Cy-3-g（纯度>96.5%）。用不同质量浓度（0、0.10、0.25μg/mL和0.50μg/mL）Cy-3-g与THP-1细胞共孵育4 h,然后加入LPS（质量浓度50 ng/mL）与细胞继续孵育48 h,用酶联免疫吸附试验法测定培养液中白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、IL-8、IL-10和肿瘤坏死因子-α(tumor necrosis fac...     

Effects of soybean ethanol extract on the prostaglandin E2 and interleukin-6 production in osteoblastic cells

We recently found that soybean ethanol extract (0.05 g/l) has a direct stimulatory effect on bone formation in cultured osteoblastic cells in vitro. The present study was conducted to investigate whether soy extract affects the prostaglandin E₂ (PGE₂) and interleukin-6 (IL-6) production of osteoblastic MC3T3-E1 cells. Stimulation with TNF-α resulted in the production of PGE₂ and IL-6 in osteoblasts. Soy extract (0.05 g/l) stimulated the constitutive PGE₂ secretion but had no significant effect on TNF-α-stimulated PGE₂ production. Also, soy extract (0.05 g/l) increased the constitutive IL-6 secretion but had no statistical significance. However, TNF-α-induced IL-6 production was increased significantly (P<0.05). These findings indicate that PGE₂ induction by soy extract in constitutive state may stimulate bone formation whereas IL-6 induction by soy extract in the presence of TNF-α may be important in the inflammatory bone diseases.     

Anti-inflammatory and proliferative responses in human and ovine ovarian surface epithelial cells

The majority of ovarian cancers (>90%) are believed to derive from the ovarian surface epithelium (OSE); a single layer covering the entire surface of the ovary. At ovulation, the OSE cell layer undergoes an inflammatory response, involving cell death and growth, in order to overcome ovarian surface rupture. Abnormalities during these processes are believed to contribute to the development of tumours. Using primary cultures of OSE cells, we have compared anti-inflammatory and proliferative responses directly between human and ovine OSE cells to further establish the use of ovine OSE cells as a suitable model system for the study of human OSE cells. In order to compare effects of inflammatory stimulation, expression and activity of 11betahydroxysteroid dehydrogenase (11betaHSD) type 1 was measured in OSE cells in response to interleukin (IL)-1alpha. As previously identified in human OSE cells, treatment of ovine OSE cells with IL-1alpha stimulated a concomitant increase of 11betaHSD type 1 mRNA (31-fold; P <0.05) and oxoreductase activity, indicating an increased production of anti-inflammatory cortisol. To compare the growth of human and ovine OSE cells, OSE cell number was measured in response to treatment with gonadotropins or growth factors. In the presence of FSH, LH or human chorionic gonadotropin (hCG), ovine and human OSE cell growth was similarly stimulated >1.2-fold (P <0.05). In the presence of connective tissue growth factor (CTGF) and more significantly insulin growth factor I (IGF-I), human and ovine OSE cell growth was also similarly stimulated >1.2-fold (P <0.05) and >1.5-fold (P <0.01), respectively. The induction of both human and ovine OSE cell growth by IGF-I or hCG was further shown to be dependent on activation of the MAP kinase/extracellular-signal-regulated kinase (ERK) pathway. Stimulation of ovine OSE cell growth by hepatocyte growth factor (HGF) was similarly shown to be ERK-dependent; however, for human OSE cells, HGF only mildly stimulated ERK phosphorylation and failed to stimulate OSE cell growth. The demonstration that human and ovine OSE cells share similarities at the level of cell signalling, gene expression and cellular growth supports the use of ovine OSE cells as a suitable model for the study of human OSE cells.     

Persimmon peel extract attenuates PDGF-BB-induced human aortic smooth muscle cell migration and invasion through inhibition of c-Src activity

The unregulated migration and invasion of human aortic smooth muscle cells (HASMCs) into the intima is a crucial step in the development of atherosclerosis. Recently, the oriental persimmon extract (Diospyros kaki Thunb. cv. Fuyu) has been investigated for its anti-atherogenic properties, but the molecular mechanisms involved remain unclear. We investigated the inhibitory effects of persimmon peel and flesh extract on the platelet-derived growth factor (PDGF) BB-induced MMP-1 expression using Western blot, and abnormal migration and invasion of HASMCs using a modified Boyden chamber assay and a wound healing assay. We also evaluated the inhibitory effects of persimmon peel extract on aortic vessel thickening using a rat aortic sprouting assay. Persimmon peel (PPE), but not flesh extract (PFE), inhibited PDGF-BB-induced MMP-1 expression, cell migration and invasion in HASMCs, while suppressing the rat aortic sprouting. Western blot and in vitro kinase assay data demonstrated that PPE inhibited Src kinase activity and subsequently attenuated PDGF-BB-induced phosphorylation of MAPK and Akt signalling pathways. Taken together, our results indicate that persimmon peel might possess a potential anti-atherogenic effect through attenuation of ASMCs migration and invasion and aortic sprouting by direct inhibition of the c-Src kinase activity.     

蒲公英糖蛋白体外抗炎作用及对NF-κB信号通路的调控

探讨蒲公英糖蛋白(glycoprotein from Taraxacum,TG)对由脂多糖引起的RAW264.7细胞炎症的抗炎效果,并阐明其活性的基本分子机制。利用脂多糖刺激RAW264.7细胞,建立体外炎症模型,采用噻唑蓝比色法检测TG对RAW264.7细胞的增殖毒性,Griess试剂法检测了一氧化氮(nitric oxide,NO)的分泌情况,反转录聚合酶链式反应检测炎症细胞因子——白细胞介素-6(interleukin 6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)m RNA以及诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)m RNA的表达水平,酶联免疫吸附测定法测定IL-6和TNF-α的分泌量,Western blot检测P-IκB-α蛋白的表达水平,用以研究TG对核转录因子-κB(nuclear factor-kappa B,NF-κB)信号转导通路的抑制作用。结果表明:TG能够显著甚至极显著地抑制NO的分泌,IL-6、TNF-α、i NOS的m RNA的表达,IL-6和TNF-α的分泌(P0.05、P0.01)。TG高度显著上调了IκB-α的蛋白表达(P0.001),并显著下调了P-IκB-α的蛋白表达(P0.05),且与TG质量浓度成正比。其中在TG质量浓度为250、500、1 000μg/m L时对TNF-α分泌量的抑制率分别为28.6%、65.4%、89.3%,对IL-6分泌量的抑制率分别为32.3%、54.1%、85.7%。TG间接抑制了NF-κB信号转导途径,有显著的体外抗炎效果且抗炎效果与TG的质量浓度呈剂量依赖性。     

黑蒜对溃疡性结肠炎BALB/c小鼠炎症反应的影响

目的：通过葡聚糖硫酸钠（dextran sulfate sodium，DSS）诱导的BALB/c小鼠溃疡性结肠炎模型，研究黑蒜对溃疡性结肠炎小鼠炎症反应的影响。方法：小鼠自由饮水摄入质量分数4% DSS建立溃疡性结肠炎模型；分别灌胃不同剂量黑蒜水提取液；考察小鼠体质量变化、疾病活动指数（disease activity index，DAI）、病理组织学评分；苏木精-伊红染色观察小鼠结肠组织病理变化，酶联免疫吸附测定法检测小鼠血清肿瘤坏死因子α（tumor necrosis factor α，TNF-α）、白细胞介素6（interleukin 6，IL-6）、IL-1β质量浓度，免疫印迹法检测小鼠结肠组织中TNF-α的表达水平。结果：高、中、低剂量的黑蒜均缓解了DSS引起的小鼠体质量下降；与模型组相比，黑蒜高、低剂量组小鼠DAI评分显著降低（P＜0.05），黑蒜高、中剂量组小鼠病理组织学评分、IL-6、IL-1β和TNF-α质量浓度极显著降低（P＜0.01）；结论：黑蒜具有降低BALB/c小鼠溃疡性结肠炎炎症反应的作用。     

柚叶总黄酮对LPS所致Caco-2结肠上皮细胞间高通透性的保护作用

探讨柚叶总黄酮(PLTFE)对脂多糖(LPS,2 μg/mL)诱发Caco-2高通透性细胞模型的保护作用。Caco-2模型细胞以不同浓度的PLTFE(0、10、50、100、150 μg/mL)处理培养24 h进行后续实验。MTT法测定细胞生存率,细胞乳酸脱氢酶(lactate dehydrogenase,LDH)水平依说明书使用试剂盒测定。酶联法(enzyme linked immunosorbent assay,ELISA)测定白介素(interlukin-1β,IL-1β)、IL-8和肿瘤坏死因子(tumor necrosis factor-α,TNF-α)分泌水平。跨上皮细胞电阻(trans epithelial electrical resistance,TEER)值和异硫氰酸荧光素-右旋糖酐(FD40)透过度用于评估细胞通透性水平。实时定量PCR(Quantitative real-time PCR,qRT-PCR)检测细胞IL-1β、IL-8、TNF-α、闭锁蛋白(Occludin)、紧密连接蛋白-1(claudin-1)、封闭小带蛋白(ZO-1)和肌球蛋白轻链激酶(myosin light chain kinase,MLCK)的mRNA表达。结果表明,柚叶总黄酮能显著提高受损细胞生存率至86.1%,抑制受损细胞中LDH的溢出(p<0.05)。同时还能有效抑制受损细胞中炎性细胞因子(IL-1β、IL-8、TNF-α)的分泌及mRNA转录。此外,柚叶总黄酮可增强细胞紧密连接因子(Occludin、claudin-1、ZO-1)的mRNA转录,抑制MLCK的mRNA转录,改善细胞间高通透性qRT-PCR法检测细胞中相关因子的mRNA转录水平。结果提示,柚叶总黄酮具有较强的抗炎活性,能通过上调细胞内细胞紧密连接相关因子的mRNA转录显著改善LPS造成的Caco-2细胞间高通透性的发生(p<0.05)。     

Anti-inflammatory effect of the extract from fermented <Emphasis Type="Italic">Asterina pectinifera</Emphasis> with <Emphasis Type="Italic">Cordyceps militaris</Emphasis> mycelia in LPS-induced RAW264.7 macrophages

In our previous work, Asterina pectinifera was fermented with Cordyceps militaris mycelia to improve its bioactivities and was reported to have strong antioxidant activities. The aim of the current study was to investigate its anti-inflammatory effect and mechanisms of action. In this study, we observed the inhibitory effect of the extract from fermented A. pectinifera with C. militaris mycelia (FACM) on nitric oxide (NO) production and its molecular mechanism in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. FACM could decrease LPS-induced NO production. Western blot analysis showed that FACM could down-regulate LPS-induced expression of inducible NO synthase without affecting cyclooxygenase-2. Moreover, FACM exhibited anti-inflammatory activity in LPS-induced RAW264.7 mouse macrophage cells through proinflammatory mediators including TNF-α and IL-6 via nuclear factor kappa B pathway. FACM inhibited LPS-induced phosphorylation of extracellular-signal-regulated kinase expression. Our results suggest that FACM may be a potential candidate for inflammation therapy by attenuating the generation of cytokines, production of NO, and generation of ROS in RAW264.7 cells.     

Anti-inflammation of epicatechin mediated by TMEM35A and TMPO in bovine mammary epithelial cell line cells and mouse mammary gland

Epicatechin (EC) has significant antiinflammation, antioxidation, and anticancer activities. It also provides a new alternative treatment for mastitis, which can result in great economic losses in the dairy industry if left untreated. The purpose of this study was to investigate the anti-inflammatory effects of EC on mastitis and the underlying mechanism using in vivo and in vitro systems. The use of ELISA and immunohistochemistry assays showed that EC treatment at 1.5, 7.5, 15, and 30 mg/mL decreased protein expression of inflammatory mediators, including cyclooxygenase-2 and inducible nitric oxide synthase; inflammatory cytokines, which were composed of IL-1β, TNF-α, and IL-6 in lipopolysaccharide (LPS)-stimulated bovine mammary epithelial cell line (MAC-T); and mouse mammary gland, together with reduced filtration of T lymphocytes in the mouse mammary gland. Furthermore, EC treatment reduced LPS-induced phosphorylation levels of p65 and inhibitor of NF-κB, and blocked nuclear translocation of p65 as revealed by western blot and immunofluorescence test in MAC-T cells and the mouse mammary gland. Epicatechin also attenuated LPS-induced phosphorylation levels of mitogen-activated protein kinase members (i.e., p38, c-Jun N-terminal kinase 1/2 and extracellular regulated protein kinases 1/2). Using RNA-seq and tandem mass tag analyses, upregulation of TMEM35A and TMPO proteins was disclosed in MAC-T cells cotreated with LPS and EC. Although clustered regularly interspaced short palindromic repeats/Cas9-based knockdown of TMEM35A and TMPO attenuated abundance of phosphorylated (p)-p65, p-p38, TNF-α, and iNOS, overexpression of TMEM35A reversed EC-mediated effects in TMPO knockdown cells. Moreover, interaction between TMEM35A and TMPO was detected using the co-immunoprecipitation method. In conclusion, our data demonstrated that EC inhibited LPS-induced inflammatory response in MAC-T cells and the mouse mammary gland. Importantly, TMEM35A mediated the transmembrane transport of EC, and the interaction between TMEM35A and TMPO inhibited MAPK and NF-κB pathways.     

Effects of vitamin D3 on the expression of growth-related oncogene-α in THP-1 cells and human primary monocytes

Asthma and many autoimmune diseases, such as systemic lupus erythematosus, have been reported to associate with vitamin D deficiency recently. Growth-related oncogene-α (GRO-α)/CXCL1, a neutrophil-related chemokine, have an important influence on the chronic inflammation of these diseases. It is unknown whether vitamin D has regulatory effects on GRO-α expression in human monocytes. To this end, the human monocytic leukemia cell line, THP-1, and human primary monocytes were pretreated with 1α, 25-(OH)(2)D(3), and was stimulated with lipopolysaccharide (LPS). Supernatants were collected to determine GRO-α level by ELISA. The intracellular signaling was investigated by nuclear factor (NF)-κB inhibitor, the mitogen-activated protein kinase (MAPK) inhibitors, and Western blot. In our studies, LPS-induced GRO-α was significantly enhanced in THP-1 cells, but suppressed in human primary monocytes by 1α, 25-(OH)(2)D(3). Western blotting revealed that 1α, 25-(OH)(2)D(3) increased LPS-stimulated pp38 expression in THP-1 cells, but suppressed LPS-stimulated pMEK1/2-pERK and pJNK in human primary monocytes. In conclusion, the opposite effects of 1α, 25-(OH)(2)D(3) on GRO-α expression in THP-1 cells and human primary monocytes indicated that the data from THP-1 cells should be further confirmed by human primary monocytes. Moreover, vitamin D3 may have potentiality in treating GRO-α-related chronic inflammatory diseases, like asthma and autoimmune diseases.     

红汁乳菇中愈创木烷型倍半萜化合物的抗炎活性

以新鲜红汁乳菇子实体为原料,采用有机溶剂浸提提取、乙酸乙酯萃取、硅胶柱层析得到单体化合物,通过紫外-可见光谱、高效液相色谱、质谱和核磁共振波谱分析鉴定单体化合物的结构,利用脂多糖（lipopolysaccharide,LPS）诱导RAW264.7巨噬细胞炎症模型,通过聚合酶链式反应和免疫印迹法分析单体化合物对炎症因子mRNA和蛋白相对表达水平的影响,以及对丝裂原活化蛋白激酶（mitogen-activated protein kinase,MAPK）炎症信号通路的作用。结果：从红汁乳菇中提取出具有较强抗炎活性的单体化合物,经鉴定为愈创木烷型倍半萜类化合物;与模型组相比,该倍半萜能极显著降低脂多糖刺激巨噬细胞中白细胞介素-6（interleukin,IL）-6、IL-1β、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）和一氧化氮合酶（inducible nitric oxide synthase,iNOS）的mRNA相对表达水平（P＜0.01）;该倍半萜能降低LPS刺激的巨噬细胞中炎症因子环氧化酶-2（cyclooxygenase-2,COX-2）、IL-1β、IL-6、iNOS和TNF-α蛋白相对表达水平,且呈现一定的浓度依赖性;该倍半萜能降低磷酸化的细胞外信号激酶（p44/42）、p38 MAPK（简称p38蛋白）和c-Jun氨基端激酶（c-Jun N-terminal kinase,JNK）3 种蛋白激酶的磷酸化水平,从而抑制MAPK炎症通路的活性。     

山楂果胶低聚半乳糖醛酸提取物对中波紫外线辐射HaCaT细胞氧化损伤和光老化的保护作用

探讨了山楂果胶低聚半乳糖醛酸提取物对中波紫外线（ultraviolet B,UVB）辐射后人永生化表皮角质形成细胞（HaCaT）的氧化损伤和光老化的保护作用。体外培养HaCaT细胞,分为对照组（HaCaT细胞不照射UVB,不给予山楂果胶低聚半乳糖醛酸提取物处理）、UVB辐射组（30 mJ/cm2）、低剂量（5 μg/mL）山楂果胶低聚半乳糖醛酸＋30 mJ/cm2 UVB辐射组、高剂量（10 μg/mL）山楂果胶低聚半乳糖醛酸＋30 mJ/cm2 UVB辐射组。以噻唑蓝比色法检测细胞活力,采用试剂盒检测超氧化物歧化酶（superoxide dismulase,SOD）、谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-Px）、过氧化氢酶（catalase,CAT）和乳酸脱氢酶（lactate dehydrogenase,LDH）活力,丙二醛（malondialdehyde,MDA）含量,羟脯氨酸（hydroxyproline,Hyp）质量浓度和活性氧（reactive oxygen species,ROS）水平。采用酶联免疫吸附测定实验检测肿瘤坏死因子-α（tumor necrosis factor alpha,TNF-α）、白细胞介素-6（interleukin-6,IL-6）质量浓度和基质金属蛋白酶-1（matrix metalloproteinase 1,MMP-1）含量。结果表明,经UVB照射后,HaCaT细胞活力明显降低。低、高剂量的山楂果胶低聚半乳糖醛酸提取物对经UVB辐射的HaCaT细胞具有显著的增殖作用（P＜0.05）;SOD、GSH-Px、CAT活力增加;LDH活力、ROS水平和MDA含量下降;Hyp质量浓度增加;MMP-1含量和TNF-α、IL-6质量浓度下降,且与辐射组比较均具有显著或极显著差异（P＜0.05或P＜0.01）。实时荧光定量聚合酶链式反应检测MMP-1、TNF-α及IL-6 mRNA表达水平与其含量或质量浓度检测结果一致。因此,山楂果胶低聚半乳糖醛酸对UVB辐射HaCaT细胞的氧化损伤及光老化具有保护作用。