调心方对Aβ大鼠痴呆模型脑组织IL-1β、IL-6及APPmRNA表达的影响

目的 探讨中药治疗Alzheimer型痴呆(AD)的有效方剂调心方的神经免疫调节作用机制。方法 采用免疫组化、RT-PCR方法分别研究了侧脑室注射β-淀粉样蛋白(Aβ1-40)诱导的AD大鼠模型神经免疫学病理变化。主要包括脑组织中胶质细胞原纤维酸性蛋白(GFAP)的表达及炎性细胞因子白细胞介素-β3(IL-1β)、白细胞介素-6(IL-6)mRNA和AB前体BAPPmRNA基因表达水平,并观察调心方的作用。结果 Aβ在脑组织的沉积能诱导星形胶质细胞的激活以及炎性细胞因子IL-1β、IL-6mRNA水平异常增高,APPmRNA水平也明显提高;而调心方能有效控制以上病理变化。结论 调心方能有效地控制Aβ启动的炎症和免疫级联反应,提示有效控制AD患者脑内Aβ免疫级联反应是调心方的重要作用机制之一。     

芫花总黄酮的抗炎机制研究

目的研究芫花总黄酮（TFDG）对脂多糖（LPS）协同干扰素γ（IFN-γ）诱导鼠RAW264.7巨噬细胞炎症模型中亚硝酸盐含量、诱导型合酶和细胞因子表达的影响,并从ERK/MAPK通路探讨其抗炎机制。方法细胞随机分为芫花总黄酮处理组,阳性药物组（I-NIL50μM）、炎症模型组和正常对照组;Griess反应测定TFDG25、50、100mg/L3个剂量和预刺激、同时加入、预处理3个处理时间及I-NIL对激活细胞上清液中亚硝酸盐的影响,并测定TFDG3个剂量及I-NIL对静息细胞上清液中亚硝酸盐产量的影响;MTT法测定TFDG3个剂量和I-NIL分别对激活细胞和静息细胞的细胞活力的影响;RT-PCR检测TFDG50、100mg/L预处理1h对激活细胞中诱导型合酶iNOS、COX-2、细胞因子IL-1β、IL-6、TNF-α基因表达的影响;Western blot检测TFDG50、100mg/L预处理1h对激活细胞中诱导型合酶iNOS、COX-2和p-ERK蛋白表达的影响。结果模型组中一氧化氮（NO）产物亚硝酸盐含量、诱导型合酶iNOS和COX-2基因和蛋白表达,细胞因子IL-1β、IL-6、TNF-α基因表达,ERK蛋白磷酸化水平均明显高于正常对照组（P〈0.01,P〈0.05）,细胞活力显著下降（P〈0.01）;TFDG呈剂量和时间依赖性抑制激活细胞上清液中亚硝酸盐含量（P〈0.01）,提高炎症损伤的细胞活力（P〈0.01）;TFDG对静息细胞上清液中亚硝酸盐含量无影响,无细胞毒性且于100mg/L剂量促进细胞增殖（P〈0.05）。阳性药物I-NIL于50μM剂量的抑制率为35.20%（P〈0.01）,提高炎症损伤后的细胞活力（P〈0.01）,但对静息细胞呈细胞毒性（P〈0.05）。与模型组比较,芫花总黄酮下调iNOS基因和蛋白表达（P〈0.01,P〈0.05）,抑制COX-2基因表达（P〈0.01）,对COX-2蛋白表达仅在高剂量100mg/L具抑制效果（P〈0.01）;下调细胞因子IL-1β、IL-6、TNF-α的基因表达（P〈0.01）,同时能抑制ERK蛋白的磷酸化水平（P〈0.05）。结论芫花总黄酮抑制激活细胞中iNOS基因和蛋白的表达从而降低NO稳定产物亚硝酸盐的产量,抑制COX-2基因和蛋白的表达,同时抑制细胞因子IL-1β、IL-6、TNF-α的基因表达,部分机制为抑制ERK/MAPK信号通路中ERK蛋白磷酸化而达到多靶点抗炎效果。     

The role of TGF-beta 1 and cytokines in the modulation of liver fibrosis by Sho-saiko-to in rat's bile duct ligated model

Liver fibrosis is an over-accumulation of extra-cellular matrix (ECM) and the hepatic stellate cell (Ito cell) play a central role in the pathogenesis of liver fibrosis. There are a lot of growth factors and cytokines involved in the activation of hepatic stellate cell, including of transforming growth factor (TGF-alpha, TGF-beta1), platelet-derived growth factor (PDGF), interleukin (IL-1alpha,beta, IL-6) and tumor necrosis factor (TNF-alpha). Sho-saiko-to (TJ-9; Xiao-Chai-Hu-Tang in Chinese) was the most popular herbal medicine for the treatment of chronic liver disease in Chinese and Japanese. Our aim of the current study was to examine whether TJ-9 regulated the growth factors and cytokines in the fibrogenesis of bile duct ligated model. Therefore, we assessed the TJ-9's potential in regulating TGF-beta1, PDGF mRNA expression, the amount of IL-1alpha, IL-1beta, IL-6, TNF-alpha and the fibrotic marker "PIII NP" in the serum. Then, using the immunohistochemical stain to observe the TGF-beta1 expression in the tissue. Our results showed that TJ-9 at a dose of 0.5 g/(kgday) significantly reduced the serum level of PIII NP, the mRNA expression of TGF-beta1 and PDGF. For the cytokines involved in the activation of Ito cell, TJ-9 at a dose of 0.5 g/(kgday) significantly suppressed the increasing tendency of IL-1beta and enhanced the production of TNF-alpha. Finally, we concluded that: (1) TJ-9 at a dose of 0.5g/(kgday) significantly reduced the serum fibrotic marker PIII NP in the bile duct ligated model, and its mechanism was partly by means of downregulating the mRNA of TGF-beta1 and PDGF. These results also confirmed by the immunohistochemical staining of TGF-beta1. (2) TJ-9 at a dose of 0.5 g/(kgday) suppressed the increasing tendency of IL-1beta and stimulated the production of TNF-alpha to inhibit Ito cell proliferation and collagen formation.     

Effect of Seabuckthorn (Hippophae rhamnoides) flavone on immune system: an in-vitro approach

There are several reports, which suggest that the consumption of foods rich in flavonoids is associated with a lower incidence of certain degenerative diseases, including cardiovascular disease. Flavones, of Seabuckthorn (SBT) (Hippophae rhamnoides L.) fruit berry can modulate the production and level of several signaling molecules associated with immune function and inflammation in vitro, including several cytokines. We have evaluated the immunomodulatory activity of ethanolic solution of SBT flavone (FLV) in human peripheral blood mononuclear cells (PBMCs). The SBT flavone was found to stimulate production of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in PBMCs. However, increased expressions of p-IkappaB, NF-kappaB, and p-p38 were found in flavone-treated human PBMCs with significantly suppressed expression of CD25 (IL-2R). There was no alteration found in the nitric oxide (NO) production in mouse macrophage cell line RAW 264.7. These observations suggest that stimulation of IL-6 and TNF-alpha secretion may contribute to the putative beneficial effects of dietary flavone against microbial infection.     

Treatment of THP-1 cells with Uncaria tomentosa extracts differentially regulates the expression if IL-1beta and TNF-alpha

Uncaria tomentosa, commonly known as cat's claw, is a medicinal plant native to Peru, which has been used for decades in the treatment of various inflammatory disorders. Uncaria tomentosa can be used as an antioxidant, has anti-apoptotic properties, and can enhance DNA repair, however it is best know for its anti-inflammatory properties. Treatment with Uncaria tomentosa extracts inhibits the production of the pro-inflammatory cytokine, TNF-alpha, which is a critical mediator of the immune response. In this paper, we showed that treatment of THP-1 monocyte-like cells with Uncaria tomentosa extracts inhibited the MAP kinase signaling pathway and altered cytokine expression. Using ELISA assays, we showed that treatment with Uncaria tomentosa extracts augmented LPS-dependent expression of IL-1beta by 2.4-fold, while inhibiting the LPS-dependent expression of TNF-alpha by 5.5-fold. We also showed that treatment of LPS-stimulated THP-1 cells with Uncaria tomentosa extracts blocked ERK1/2 and MEK1/2 phosphorylation in a dose-dependent manner. These data demonstrate that treatment of THP-1 cells with Uncaria tomentosa extracts has opposite effects on IL-1beta and TNF-alpha secretion, and that these changes may involve effects on the MAP kinase pathway.     

Effect of electroacupuncture on the expression of interlukin-1beta mRNA after transient focal cerebral ischemia

It has been reported that interleukin-1beta (IL-1beta ) play a key role in the pathogenesis of cerebral ischemia. Acupuncture is an effective traditional medical therapy in China. The aim of present study was to evaluate the effect of electroacupuncture (EA) on IL-1beta mRNA expression after middle cerebral artery occlusion (MCAO) in rats. Using in situ hybridization technique, it was found that in the MCAO group the expression of IL-1beta mRNA was significantly increased at 2h, 6h, 12h after reperfusion in cerebral ischemic cortex compared with normal group. In EA+ MCAO group the expression of IL-1beta mRNA was significantly decreased at 2h, 6h and 12h in ischemic cortex compared with MCAO group. The results indicated that EA might decrease the IL-1beta protein expression by reducing the IL-beta mRNA expression in ischemic cortex.     

Therapeutic effect of Korean red ginseng on inflammatory cytokines in rats with focal cerebral ischemia/reperfusion injury

This study aims to identify the therapeutic effect of Korean red ginseng (KRG) on the expression of inflammatory cytokines in rats with focal cerebral ischemia/reperfusion injury. Adult male Sprague-Dawley rats were subjected to transient middle cerebral artery occlusion (tMCAO) for two hours. They were fed KRG extract (100 mg/kg/day per orally) or saline after reperfusion. Tests for neurological deficits, using the modified neurologic severity score and the corner turn test, were performed before the ischemic event, and one, three, and seven days after tMCAO. Serum levels of cytokines were measured three and seven days after the operation, using enzyme-linked immunosorbent assays. The infarct volume was assessed after seven days by staining brain tissue with 2% 2, 3, 5-triphenyltetrazolium chloride. Oral administration of KRG significantly reduced the infarct volumes and rapidly improved neurological deficits. Serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and IL-6 were higher in tMCAO-operated rats than in the sham-operated rats. These changes were attenuated by daily KRG intake for seven days. Serum IL-10 levels were significantly increased in KRG-fed rats, as compared to sham-operated and saline-fed rats. Our results suggested that KRG provides neuroprotection for rats with focal cerebral ischemia/reperfusion injury. This neuroprotection may be due to raised IL-10 expression and a reduction in the serum levels of TNF-α, IL-1β, and IL-6.     

Antiviral and immunomodulatory effect of a lyophilized extract of Capparis spinosa L. buds

Herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) are common human pathogens that in particular cases can also cause severe problems especially in immunodeficient patients. The present paper reports the antiviral and immunomodulatory properties of a methanolic extract of C. spinosa buds (CAP), rich in flavonoids, including several quercetin and kaempferol glycosides. In particular we have investigated whether the in vitro exposure of human peripheral blood mononuclear cells (PBMCs) to CAP might inhibit the replication of HSV-2 and modulate the induction kinetics of IL-12, TNF-alpha IFN-gamma. Our findings have shown that CAP treatment interferes with HSV-2 replication in PBMCs inhibiting the extracellular virus release upregulating their production of IL-12, IFN-gamma and TNF-alpha. One could speculate that CAP may contribute in improving immune surveillance of PBMCs toward virus infection by up-regulating expression of peculiar proinflammatory cytokines; it should thus be successfully employed for treatment of HSV-2 infections in immunocompromised hosts.     

板蓝根多糖对内毒素诱导人单核/巨噬细胞炎症因子表达的影响

目的研究板蓝根多糖（IIP）对内毒素诱导的人单核/巨噬细胞中多种炎症因子表达的影响。方法采用体外培养的人单核/巨噬细胞分为空白对照组（Control组）、内毒素（LPS）组、板蓝根多糖＋LPS组（IIP＋LPS组）,以100ng/ml内毒素作为刺激因子,分别加以相应的干预。应用人炎症因子抗体芯片检测各组人单核/巨噬细胞中IL-1β、IL-6、IL-8和金属蛋白酶组织抑制剂-2（TIMP-2）等炎症因子表达的变化情况。结果与Control组比较,LPS组中IL-1β、IL-6、IL-8的表达增强,而IIP＋LPS组中IL-1β、IL-8、TIMP-2的表达增强。与LPS组比较,IIP＋LPS组的IL-1β、IL-6的表达降低,TIMP-2蛋白升高,差异有统计学意义（P〈0.01）。结论板蓝根多糖对内毒素诱导的人单核/巨噬细胞多种炎症因子表达有调节作用。     

In vitro anti-inflammatory effects of arctigenin,a lignan from Arctium lappa L., through inhibition on iNOS pathway

Ethnopharmacological relevance

Arctigenin, a bioactive constituent from dried seeds of Arctium lappa L. (Compositae) which has been widely used as a Traditional Chinese Medicine for dispelling wind and heat included in Chinese Pharmacophere, was found to exhibit anti-inflammatory activities but its molecular mechanism remains unknown yet.

Aim of the study

To investigate the anti-inflammatory mechanism of arctigenin.

Materials and methods

Cultured macrophage RAW 264.7 cells and THP-1 cells were used for the experiments. Griess assay was used to evaluate the inhibitory effect of arctigenin on the overproduction of nitric oxide (NO). ELISA was used to determine the level of pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6). The inhibitory effect on the enzymatic activity of cyclooxygenase-2 (COX-2) was tested by colorimetric method. Western blot was used to detect the expression of inducible nitric oxide synthase (iNOS) and COX-2.

Results

Arctigenin suppressed lipopolysaccharide (LPS)-stimulated NO production and pro-inflammatory cytokines secretion, including TNF-α and IL-6 in a dose-dependent manner. Arctigenin also strongly inhibited the expression of iNOS and iNOS enzymatic activity, whereas the expression of COX-2 and COX-2 enzymatic activity were not affected by arctigenin.

Conclusions

These results indicated that potent inhibition on NO, TNF-α and IL-6, but not COX-2 expression and COX-2 activity, might constitute the anti-inflammatory mechanism of arctigenin. Arctigenin suppressed the overproduction of NO through down-regulation of iNOS expression and iNOS enzymatic activity in LPS-stimulated macrophage.     

Inhibition of lipopolysaccharide plus substance P-induced tumor necrosis factor-alpha production from astrocytes by Chongmyung-Tang.

Astrocytes have the capacity to secrete or respond to a variety of cytokines. In this study, we have examined whether an aqueous extract of Chongmyung-Tang (CmT) inhibits production of tumor necrosis factor-alpha (TNF-alpha) from primary cultures of mouse astrocytes. CmT (1 and 10 microg/ml) significantly inhibited the TNF-alpha production by astrocytes stimulated with lipopolysaccharide (LPS) and substance P (SP). Interleukin-1 (IL-1) has been shown to elevate TNF-alpha production from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore examined whether IL-1 mediated inhibition of TNF-alpha production from primary astrocytes by CmT. Treatment of CmT (1 and 10 microg/ml) to astrocytes stimulated with both LPS and SP decreased IL-1 production significantly. In addition, reverse-transcribed polymerase chain reaction analysis demonstrated that significantly reduced level of the TNF-alpha mRNA was expressed in astrocytes treated with CmT. Our results suggest that CmT inhibits TNF-alpha production by reducing TNF-alpha mRNA expression.     

The Effect of propolis on Th1/Th2 cytokine expression and production by melanoma-bearing mice submitted to stress

Since propolis possesses immunomodulatory and antitumoral activities, this work aimed to evaluate its effect on Th1 (IL-2 and IFN-γ) and Th2 (IL-4 and IL-10) cytokines mRNA expression and production by melanoma-bearing mice submitted to immobilization stress. C57BL/6 male mice were inoculated with B16F10 cells, treated with propolis and submitted to stress for 14 days. Spleen cells were assessed for Th1/Th2 cytokine expression and production. Stress induced a higher tumor area, while propolis-treated mice, stressed or not, showed a melanoma development similar to the control. In groups without melanoma, stress or propolis treatment did not affect IL-2, IL-4 and IL-10 gene expression. On the other hand, IL-2 and IL-10 expression was inhibited in melanoma-bearing mice, stressed or not. Th1 cytokine production was also inhibited in melanoma-bearing mice. Propolis administration to melanoma-bearing mice submitted to stress stimulated IL-2 expression, as well as Th1 cytokine (IL-2 and IFN-γ) production, indicating the activation of antitumor cell-mediated immunity. Propolis also stimulated IL-10 expression and production, which may be related to immunoregulatory effects. The data indicate that propolis exerted an immunomodulatory activity in this assay, which may be related to its antitumoral action in vivo.     

Protective effect of Astragali radix extract on interleukin 1beta-induced in fl ammation in human amnion

The aim of this study was to investigate the effect of Astragali radix extract on interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha production and prostaglandin E(2) (PGE(2)) and leukotriene C(4) (LTC(4)) release from IL-1beta-stimulated human amnion. Primary monolayer cultures of amnion cells were established from women undergoing elective caesarean section before the onset of labour. Production of both IL-6 and TNF-alpha was stimulated in a concentration-dependent manner by proinflammatory cytokine IL-1beta (0.01-10 ng/mL). Astragalus extract inhibited IL-6 production by approximately 75% from cells under IL-1beta-stimulated conditions. Treatment of amnion cells with IL-1beta (0.01-10 ng/mL) resulted in a significant increase in PGE(2) release. Incubation of the cells with the extract for 24 h significantly inhibited IL-1beta-induced PGE(2) production. A concentration-dependent increase in LTC(4) production by amnion cells occurred in response to IL-1beta. Astragalus extract blocked the effect of IL-1beta in LTC(4) production in human amnion. These results indicate that Astragali radix has a broad antiinflammatory effect in human amnion and may be considered a promising agent to protect preterm labour.     

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??OBJECTIVE To investigate the inhibitory effects of phenolic glycoside compound xylocontroside D on neurotoxicity of primary cultured cortical neurons induced by A??_1-42. METHODS The primary cortical neurons and microglia cells derived from rat cerebral tissues were used and neurotoxicity were induced by A??_1-42. Then the thiazolyl blue tetrazolium bromide test(MTT) was applied for detecting cell survivability; cells were incubated with CH₃-H₂DCF-DA for evaluating cellular endogenous reactive oxygen species(ROS) level by flow cytometry;fluorescent staining of 8-hydroxy-2-deoxyguanosine(8-OH-DG) was applied for identifying the metabolites of ROS based DNA damage; fluorescent imaging was applied for illustrating the activation of microglia cells and the enzyme-linked immunosorbent assay(ELISA) was applied for analysis of pro-inflammatory cytokines, cyclooxygenase-1(COX-1), cyclooxygenase-2(COX-2), interleukin-1beta(IL-1??) and tumor necrosis factor alpha(TNF-??). RESULTS Xylocontroside D showed significant inhibitory effects on A??_1-42 induced neurotoxicity, including cell death, cellular endogenous ROS levels and DNA damage caused by ROS reaction. Moreover, this compound could also prevent the activation of microglia, decrease the production or secretion of pro-inflammatory cytokines that produced by activated microglia, such as COX-1, COX-2, IL-1?? and TNF-??. CONCLUSION Our RESULTS indicated that xylocontroside D was able to inhibit the oxidative stress injury and neuroinflammation of primary cultured cortical neurons induced by A??_1-42.