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.     

竹叶黄酮对大鼠肝脏缺血再灌注损伤的影响及机制研究

目的：观察竹叶黄酮对大鼠肝脏缺血再灌注损伤的保护作用并探讨其作用机制。方法将SD大鼠按随机数字表法分为假手术组、模型组、竹叶黄酮组。竹叶黄酮组大鼠腹腔注射竹叶黄酮溶液30 mg/kg,假手术组及模型组大鼠腹腔注射等体积生理盐水,1次/d,连续注射14 d。末次给药次日,模型组及竹叶黄酮组建立大鼠肝脏缺血再灌注模型。于再灌注后6 h,检测各组大鼠血清ALT、AST水平;采用ELISA法检测各组大鼠血清IL-1、IL-6水平;采用HE染色观察各组大鼠肝组织病理学改变;采用Western Blot检测肝组织半胱氨酸天冬氨酸蛋白酶-3(caspase-3)表达。结果与模型组比较,竹叶黄酮组大鼠血清 ALT[(343.11±78.32)U/L 比(215.31±52.21)U/L]、AST[(863.97±63.58)U/L 比(501.59±70.13)U/L]、IL-1[(596.85±102.34)U/L 比(315.30±88.64)U/L]、IL-6[(183.64±20.69)U/L 比(106.75±10.25)U/L]水平降低(P＜0.05),竹叶黄酮组大鼠肝组织caspase-3[(0.38±0.05)比(0.86±0.09)]表达水平降低(P＜0.05)。结论竹叶黄酮预处理可减轻大鼠肝脏缺血再灌注损伤,其作用机制可能与抑制caspase-3蛋白表达有关。     

Immunomodulatory properties of Achyrocline satureioides (Lam.) D.C. infusion: a study on human leukocytes

AIM OF THE STUDY: Achyrocline satureioides (Lam.) D.C. is a South American native medicinal herb known by the popular name of "Marcela". Its infusion is widely utilized for the treatment of several digestive ailments, as an anti-inflammatory preparation, as a sedative and anti-atherosclerotic. Circumstantial evidence suggests that extracts of Achyrocline satureioides may have immunomodulatory properties. The present study was therefore devised to investigate the in vitro effects Achyrocline satureioides infusion on human peripheral blood mononuclear cells (PBMCs) and polymorphonuclear leukocytes (PMNs). MATERIALS AND METHODS: Experiments were performed on cells isolated from venous blood obtained from healthy donors. PBMC proliferation and cytokine production were assessed by standard ELISA methods. Reactive oxygen species (ROS) production by PMNs was evaluated by spectrofluorimetry. RESULTS: In PBMCs, Achyrocline satureioides infusion in the 0.06-0.24microg/ml quercetin equivalent (QE) concentration range concentration-dependently reduced PHA-induced proliferation and production of interferon (IFN)-gamma and interleukin (IL)-4. Lower concentrations of the infusion (0.006-0.03microg/ml QE), which were ineffective on cell proliferation, significantly increased the production of both IFN-gamma and IL-4 and decreased the ratio IFN-gamma/IL-4. In PMNs, Achyrocline satureioides infusion slightly increased the spontaneous generation of ROS only at concentrations > or =0.06microg/ml QE. On the contrary, in the 0.0012-0.03microg/ml QE concentration range the infusion profoundly inhibited fMLP-induced ROS generation as well as spontaneous and fMLP-induced IL-8 production. CONCLUSIONS: The present results provide evidence that Achyrocline satureioides infusion may exert several immunomodulatory effects, in line with its traditional use as an anti-inflammatory agent in many disease conditions. Further studies are warranted to better characterize such effects and to assess their therapeutic relevance.     

吸附树脂再生效果的实验研究

目的:对吸附树脂常规再生方法的再生效果进行评价。方法:应用D101大孔吸附树脂,对淫羊藿总黄酮进行反复吸附、洗脱10次后,按常规方法进行再生处理;然后检测再生后树脂上淫羊藿总黄酮残留量,检测再生后树脂吸附容量,观察再生后树脂的变化。结果:在实验条件下,树脂柱上有前10次吸附平均收率0.6%的淫羊藿总黄酮残留量,再生后的吸附容量为24.30 mg·ml-1。结论:吸附树脂可连续多批次地用于同一产物的提取分离,再生后有提取物残留,吸附容量变化不大,再生后有利于提高树脂柱吸附过程的流动性。     

Anti-atherogenic effect of San-Huang-Xie-Xin-Tang, a traditional Chinese medicine, in cultured human aortic smooth muscle cells

Aim of the study

San-Huang-Xie-Xin-Tang (SHXXT) is a traditional Chinese medicine and it has been shown to have an anti-inflammatory effect. Since inflammation is one of the major mechanisms of atherosclerosis, we aimed to investigate anti-atherosclerotic effect of SHXXT in human aortic smooth muscle cells (HASMCs).

Materials and methods

Human aortic smooth muscle cells (HASMCs) were used in the present study, and rendered atherosclerosis by adding lipopolysaccharides. We first tested the effects of SHXXT on HASMC migration and proliferation as they present the major morphological change of atherosclerosis. We also examined whether SHXXT can influence the production of several biomarkers of inflammation and atherosclerosis including reactive oxygen species (ROS), COX-2, ERK1/2, IL-1β, IL-6, IL-8 and MCP-1.

Results

Using the dimethyl-thiazol-diphenyltetrazoliumbromide (MTT) and wound repair assay, SHXXT was shown to significantly reduce HASMC proliferation and migration, respectively. From the fluorometric assay, SHXXT significantly reduced ROS production. SHXXT down regulated mRNA and protein levels for the COX-2 gene. In addition, phosphorylated ERK1/2 levels were suppressed by SHXXT suggesting HASMC division can be inhibited under pro-inflammatory condition. SHXXT significantly inhibited the production of IL-1β, IL-6, IL-8 and MCP-1 after LPS stimulation.

Conclusions

Our results indicated that SHXXT can influence several mechanisms involved in atherosclerosis, which suggests that SHXXT may have a therapeutic potential for cardiovascular disease associated with atherosclerosis.     

Protection of Total Flavonoid Fraction from Nervilia fordii on Lipopolysaccharide-induced Acute Lung Injury in Rats

Objective To investigate the effects of total flavonoid fraction(TFF) from Nervilia fordii on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats,and to explore their protective mechanism.Methods LPS-induced ALI model was established by LPS(5 mg/kg) injection via left cervical vein.Blood samples were collected from the cervical artery of all rats at 5 and 6 h after LPS challenge for arterial blood gas test and cytokines measurements,and pulmonary microvascular permeability(PMP),lung wet/dry weight ratio(W/D),and pathological features were observed.Results Phytochemical study showed that the TFF contained 67.3% of flavonoids expressed in rutin and three flavone glycosides.The TFF pretreatment(6.24 and 12.48 mg/kg) attenuated the partial arterial pressure of oxygen decline in blood significantly,and decreased the PMP and lung W/D in ALI rats.In addition,the TFF(6.24 and 12.48 mg/kg) also ameliorated the LPS-induced lung damages including alveolar edema,neutrophils infiltration,alveolar hemorrhage,and thickening of the alveolar wall.Furthermore,the treatment with the TFF(6.24 and 12.48 mg/kg) also down-regulated the levels of pro-inflammatory cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and intercellular adhesion molecule-1(ICAM-1),and up-regulated the level of anti-inflammatory cytokine IL-10 in serum of ALI rats simultaneously.Conclusion These results suggest that the TFF could protect LPS-induced ALI in rats,which may be mediated,at least in part,by adjusting the production of inflammatory cytokines including TNF-α,IL-6,ICAM-1,and IL-10.     

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.     

Potential anti-inflammatory effects of Melaleuca alternifolia essential oil on human peripheral blood leukocytes

The fungicidal and bactericidal actions of the essential oil (EO) of Melaleuca alternifolia seem well established, but their anti-inflammatory and antioxidative effects remain unclear. This study investigated in vitro the possible role of whole Melaleuca alternifolia EO as a modulator of the inflammatory/non-specific immune response by exploring the chemotaxis and kinetic radical oxygen species (ROS) production of leukocytes and cytokine secretion in peripheral blood mononuclear cells (PBMCs) in humans. The influence of Melaleuca alternifolia EO on the chemotaxis under agarose of isolated neutrophils (PMNs) was evaluated. The kinetics of ROS production by stimulated total circulating leukocytes was followed over 2 h by recording the fluorescence intensity of oxidized dihydrorhodamine 123. The effects of this EO on pro-(interleukin IL-2) and anti-(IL-4 and IL10) inflammatory cytokine secretions were determined by ELISA following incubation of PBMCs with the EO for 24 h. Melaleuca alternifolia EO was inefficient on the chemotaxis of PMNs. It exerted an antioxidant effect, reducing ROS production throughout the kinetic study. Melaleuca alternifolia EO inhibited PBMC proliferation, as revealed by a reduction in IL-2 secretion by stimulated lymphocytes. This EO at 0.1% directly increased the secretion of the anti-inflammatory cytokine IL-4 compared with IL-4 secretion without EO (18.5 +/- 10.0 vs 3.3 +/- 1, p < 0.05), and also increased IL-10 secretion at 0.01% (94.9 +/- 38.7 vs 44.1 +/- 18, ns). Melaleuca alternifolia EO may not only act as an anti-inflammatory mediator through its antioxidant activity but may also efficiently protect the organism by reducing the proliferation of inflammatory cells without affecting their capacity to secrete anti-inflammatory cytokines.     

大黄素对人胚肺成纤维细胞的影响

目的:探讨大黄素在抗肺纤维化中的作用。方法:采用体外细胞培养、免疫组化、透射电镜和同位素检测等技术,比较大黄素作用下人胚肺成纤维细胞(LF)的增殖和其分泌的白细胞介素(IL)-6的水平,检测大黄素作用下人胚 LF 电镜下的形态学特征。结果:(1)大黄素能够抑制人胚 LF 的增殖,并与其浓度、作用时间有关,大黄素高浓度组的抑制效果明显优于低浓度组,并随着时间延长其作用也明显增强;(2)在 IL-1诱导时,大黄素可以降低人胚 LF 分泌 IL-6的水平;(3)大黄素组人胚 LF 形态上与对照组比较差异无显著性。结论:大黄素具有抑制人胚 LF 增殖和抑制 IL-1刺激人胚 LF 分泌 IL-6作用。     

健脾活血补肾汤体外干预AS 患者外周血单个核细胞IL-23 与IL-17 表达的实验研究

目的：观察健脾活血补肾汤体外对强直性脊柱炎（AS）患者外周血单个核细胞IL-23 与IL-17表达的影响,从而探讨健脾活血补肾汤治疗AS 的可能机制。方法：制备不同浓度健脾活血补肾汤大鼠含药血清,用ELISA 方法检测经不同浓度大鼠含药血清培养的AS 患者和健康志愿者PBMCs 上清液中IL-23与IL-17 水平。结果：AS 患者PBMCs 经72、36 g·kg-1·d-1 两组浓度大鼠含药血清培养后,与生理盐水组及18 g·kg-1·d-1 组相比,上清液中IL-23 与IL-17 水平显著下降（P<0.01）,但健脾活血补肾汤组含药大鼠血清对健康自愿者PBMCs 上清液中IL-23 与IL-17 表达水平影响均没有统计学意义。结论：健脾活血补肾汤可能通过下调炎症因子IL-23 和IL-17 的表达而起到延缓关节炎性骨质破坏的作用。     

In vitro immunopotentiating properties and tumour cell toxicity induced by Lophophora williamsii (peyote) cactus methanolic extract

Lophophora williamsii, also known as peyote, is found primarily in dry regions from Central Mexico, including the Mexican States of Nayarit, San Luis Potosí, Zacatecas, Nuevo León, Chihuahua, Coahuila and Tamaulipas, to Texas particularly in regions along Rio Grande. Peyote extracts have been associated with stimulating the central nervous system and regulating blood pressure, sleep, hunger and thirst. However, there is no evidence of any effect of peyote on the immune system or against tumour cell growth. The present study was designed to evaluate the in vitro effects of peyote methanolic extracts on some parameters of mouse and human leukocyte immunocompetence and tumour cell growth. Peyote extract (0.18-18 micro g/mL) activated nitric oxide production by murine macrophages, and stimulated up to 2.4-fold proliferation of murine thymic lymphocytes. In addition, peyote extract induced up to 1.85-, 2.29- and 1.89-fold increases in mRNA signal of IL-1, IL-6 and IL-8 by human leukocytes. Also examined were the effects of peyote extracts on murine lymphoma L5178Y-R and fi broblastoma L929, and human myeloid U937 and mammary gland MCF7 tumour cell growth using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). Peyote extracts were toxic for MCF7, L5178Y-R, U937 and L929 (18 mg/mL peyote extract caused 1.3%, 8%, 45% and 60% viability respectively) cell lines.     

Inhibitory effect on immunoglobulin E production in vivo and in vitro by Siegesbeckia glabrescens.

Elevated levels of immunoglobulin (Ig)E are associated with immediate-type allergic reactions. The effect of an aqueous extract of Siegesbeckia glabrescens (Compositae) whole plants (SGWP) on in vivo and in vitro IgE production was studied in mice. SGWP dose-dependently inhibited the active systemic anaphylaxis and serum IgE production induced by immunization with ovalbumin and Bordetella pertussis toxin absorbed to aluminium hydroxide gel. SGWP dose-dependently inhibited IL-4-dependent IgE production by lipopolysaccharide-stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, SGWP also showed an inhibitory effect on IgE production. These results suggest that SGWP has an anti-allergic activity by inhibiting IgE production from B cells.     

针灸通督法联合药物治疗缺血性卒中的疗效及对预后的影响

目的观察针灸通督法联合常规药物治疗急性缺血性卒中患者的疗效及对预后的影响及其与CD40/CD40L 系统的关系。方法收治缺血性卒中患者100 例,采用随机数字表法分为药物组和针药组,每组50例,药物组采用常规药物治疗,针药组采用针灸通督法联合常规药物治疗。治疗前后抽提患者外周血单个核细胞(PBMCs)并应用Real-time PCR 及Western blot 检测其中CD40 mRNA 及蛋白的表达水平;ELISA 法检测所收集血浆中sCD40L、IL-1?、IL-6 的水平;通过NIHSS 评分评价两组患者的疗效,同时比较两组患者治疗后3 个月不良事件发生情况。结果与药物组相比,针药组PBMCs 中CD40 mRNA 及蛋白表达水平下降更为显著(P<0.05);两组患者治疗后血浆sCD40L、IL-1?、IL-6 水平均明显降低(P<0.05),但针药组降低更为显著(P<0.05);针药组神经功能恢复情况显著优于药物组(P<0.05),治疗后3 个月不良事件发生率也明显低于对照组(P<0.05)。结论针灸通督法联合常规药物治疗能够明显提升缺血性卒中患者治疗疗效并降低治疗后不良事件的发生率,其机制与针灸通督法联合治疗能更有效地抑制CD40/CD40L 系统的活性有关。     

"Tien-Hsien liquid" can modulate antigen-stimulated cytokine production by T-cells isolated from patients with recurrent aphthous ulcerations

Recurrent aphthous ulcerations (RAU) represent a common oral mucosal disease with altered humoral and cellular immunities. Tien-Hsien liquid (THL) is an extract of Chinese medicinal herbs with immunomodulating effects. Our previous study found that THL can modulate the antigen-stimulated proliferative response of peripheral blood mononuclear cells and T-cells isolated from RAU patients. In this study, we further tested whether THL can modulate the antigen-stimulated cytokine production by T-cells isolated from RAU patients. To achieve this goal, T-cells isolated from 19 RAU patients were incubated with phytohemagglutinin (PHA), glutaraldehyde-inactivated tetanus toxoid (TT), glucosyltransferase D (GtfD), or antigens of Streptococcus mutans in the presence or absence of THL. The levels of interleukin (IL)-2, interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), IL-6, or IL-10 in the supernatants of T-cell cultures were measured by cytokine enzyme-linked immunosorbent assay (ELISA) kits. We found that THL significantly increased the PHA- or TT-stimulated TNF-alpha, IL-6, and IL-10 production by T-cells isolated from RAU patients. However, THL could also significantly decrease the TT-stimulated IL-2 production, the GtfD-stimulated IL-2, TNF-alpha, IL-6 and IL-10 production, and the S. mutans-stimulated IFN-gamma, TNF-alpha, and IL-10 production by T-cells isolated from RAU patients. These results indicate that THL can modulate the antigen-stimulated cytokine production by T-cells isolated from RAU patients. Because RAU is probably a Thl-mediated disease with elevated levels of IL-2, IFN-gamma, TNF-alpha and IL-6 in either the patient's sera or oral lesions and these increased levels of cytokines can be reduced by THL, we suggest that THL may be a potential immunoceutical agent for treatment of RAU.