Suppression of T lymphocyte activity by lupeol isolated from Crataeva religiosa

Lupeol has been shown to possess antiarthritic activity through possible suppression of the immune system. As seen in the following studies, it was found to suppress various immune factors such as the phagocytic (cell-killing) activity of macrophages, T-lymphocyte activity that included CD4+T cell mediated cytokine generation. Assessment of T cells and their intracellular content of cytokines was carried out by flow cytometric analysis in Balb/c mice. Oral administration of lupeol at doses of 12.5-200 mg/kg p.o. inhibited CD4+ T and CD8+ T cell counts and cytokines IL-2, IFN-gamma (Th1) and IL-4 (Th2). Cytometric bead array (CBA) technology was applied to carry out simultaneous measurement of multiple serum cytokines. The oral LD(0) in mice was more than 2 g/kg body weight.     

Anti-arthritic activity of a biopolymeric fraction from Euphorbia tirucalli

The present study was undertaken to investigate the anti-arthritic activity of a biopolymeric fraction (BET) from plant Euphorbia tirucalli Boiss (Euphorbiaceae). The fraction showed dose dependent anti-arthritic activity and also showed in vivo immunomodulatory capacity being a major component in inhibiting arthritis. It caused suppression of CD4(+) and CD8(+) T cells, inhibition of intracellular Interleukin-2 (IL-2) and Interferon-gamma (IFN-gamma) by flowcytometry. It inhibited vascular permeability and the migration of leucocytes at the site of the insult. The oral LD(0) in both rats and mice was more than 2000 mg/kg.     

Immunomodulatory activity of isoflavones isolated from Iris germanica (Iridaceae) on T‐lymphocytes and cytokines

The immunomodulatory activities of two isoflavones, 5,7‐dihydroxy‐6,4′‐dimethoxyisoflavone (irisolidone) (1) and 5,4′‐dihydroxy‐6,7‐methylenedioxyisoflavone (irilone) (2) isolated from Iris germanica (Iridaceae) is reported. Their influence on production of T‐lymphocytes (CD4+ and CD8+ cells) and T‐cell cytokines, namely Th1: IL‐2, IFN‐γ and Th2: IL‐4 and IL‐5 in a dose‐dependent manner was studied by flow cytometric method in Balb/c mice. Oral administration of drugs at doses of 0.1–0.8 mg/kg per oral dose showed 1 to possess stimulatory activity on T‐cells and Th1 cytokine production, while as 2 acted as an immunosuppressant for both cells and cytokines. The methylated products of 1 and 2 showed a similar trend to that of their parent compounds but their activity was drastically decreased revealing the importance of free phenolic groups for their immunomodulating activities. Copyright © 2008 John Wiley & Sons, Ltd.     

黄连素合用环孢素A对小鼠皮肤移植排斥反应的影响

目的观察黄连素合用环孢素A抗同种异体小鼠皮肤移植排斥反应的影响,探讨其作用机制。方法以BALB/c小鼠为供体,以C57BL/6小鼠为受体,采用背-背皮肤移植手术法制备模型。将受体小鼠按随机数字表法分为模型组、黄连素组、环孢素A组、联合组,每组20只;另取20只健康C57BL/6小鼠作为假手术组。造模后黄连素组小鼠腹腔注射黄连素100 mg/kg,环孢素A组小鼠腹腔注射环孢素A注射液10 mg/kg,联合组腹腔注射黄连素100 mg/kg及环孢素A注射液5 mg/kg,假手术组和模型组小鼠腹腔注射等体积0.5%羧甲基纤维素钠。1次/d,连续10 d。记录受体小鼠移植皮片的存活时间;采用ELISA法检测血浆Th1类细胞因子[IL-2、γ-干扰素(interferon-γ,IFN-γ)]和Th2类细胞因子(IL-4、IL-10)水平;采用流式细胞术检测脾脏T淋巴细胞亚群CD4+CD25+比例。结果与模型组比较,各给药组受体小鼠移植皮片的存活时间延长(P<0.01);联合组移植皮片存活时间较黄连素组或环孢素A组延长(P<0.01)。与模型组比较,联合组血浆IL-2[(11.55±3.14)pg/ml比(19.85±2.42)pg/ml]、IFN-γ[(26.41±6.20)pg/ml比(57.23±10.15)pg/ml]水平降低,IL-4[(192.45±70.12)pg/ml比(61.09±21.61)pg/ml]、IL-10[(106.79±27.83)pg/ml比(40.08±11.23)pg/ml]水平升高(P<0.05),脾脏CD4+CD25+调节性T细胞比例[(7.65±2.42)%比(3.69±0.83)%]升高(P<0.01)。结论黄连素合用环孢素A通过促使Thl向Th2细胞的偏移、诱导免疫耐受、刺激CD4+CD25+调节性T细胞的表达等途径,抑制同种异体小鼠的皮肤移植排斥反应。     

Evaluation of the immunomodulatory properties in mice and in vitro anti-inflammatory activity of cycloartane type saponins from Astragalus species

Ethnopharmacological relevance

Astragalus roots are used to treat leukemia and for their wound healing properties in Southeast Anatolia-Turkey.

Materials and methods

In vivo studies to investigate the effects of two Astragalus saponins were carried out on the immune response cytokines by using six to eight weeks old male Swiss albino mice. The production of IL-1β, TGF-1β, TNF-α, IL-2, IL-4 and IFN-γ cytokines was determined by ELISA. The spleen and lymph nodes, isolated from the mice subjects, were analyzed to realize induction of the surface antigen productions for IL-2Rα (CD25) and CD69. In addition, their effects on the targets of inflammation such as NF κB, iNOS and NAG-1 were investigated in cell-based assays.

Results

The results suggested that AST VII and Mac B had positive effect on Th1 cytokine release (IL-2 and IFN-γ), and suppression on Th2 cytokine production (IL-4). The immunohistochemical results exhibited induction of both IL-Rα (CD25) and CD69 surface receptors justifying the Th1 cytokine release. The compounds did not affect NF-κB or NAG-1 activity but iNOS activity was inhibited by Mac B with an IC₅₀ of 156 μg/ml.

Conclusions

The results show that Ast VII and Mac B create powerful immunoregulatory effects without the stimulation of inflammatory cytokines in mice, and have no significant effect on the inflammatory cellular targets in vitro.     

艾灸“肺俞”“膏肓”对博莱霉素A5诱导肺纤维化大鼠Ⅰ/Ⅱ型细胞因子平衡调节的影响

目的:研究艾灸“肺俞”“膏肓”对博莱霉素(BLM)所致肺纤维化大鼠肺组织Ⅰ/Ⅱ型细胞因子平衡调节的影响,探索针灸阻抑肺纤维化的效应机制。方法:SD大鼠随机分为:空白组、模型组、艾灸组、泼尼松组。气管内注入博莱霉素制作大鼠肺纤维化模型,造模后7 d开始治疗,以5 mg左右艾绒灸其双侧“肺俞”“膏肓”,每穴3壮,每天1次,10 d为1疗程,疗程间休息1 d,3个疗程后处死动物,采用酶联免疫吸附法测定肺组织内Ⅰ型细胞因子IL-2I、L-12和Ⅱ型细胞因子IL-4、IL-5含量。结果:造模后大鼠肺组织内Ⅰ型细胞因子含量降低,Ⅱ型细胞因子含量升高(均P<0.01);艾灸及泼尼松治疗能够使大鼠肺组织内Ⅰ型细胞因子含量升高,Ⅱ型细胞因子含量降低,与模型组比较具有统计学差异(P<0.05,0.01);艾灸组与泼尼松组比较,各指标均无统计学差异(P>0.05)。结论:艾灸“肺俞”“膏肓”能升高BLMA5诱导肺纤维化大鼠肺组织中Ⅰ型细胞因子IL-2I、L-12的含量,降低Ⅱ型细胞因子IL-4I、L-5的含量,提示调节Ⅰ/Ⅱ型细胞因子的平衡可能是艾灸阻抑肺纤维化发生的机理之一。     

Herbal medicine Gamgungtang down-regulates autoimmunity through induction of TH2 cytokine production by lymphocytes in experimental thyroiditis model

The crude herbal formulation, Gamgungtang (GGT), has been shown to protect animals against a wide range of spontaneously developing or induced autoimmune diseases. We have previously reported that GGT shows marked down-regulation of several experimental autoimmune diseases. Although very effective at preventing thyroid infiltrates in mice immunized with mouse deglycosylated thyroglobulin and complete Freund's adjuvant and in spontaneous models of thyroiditis, it completely failed to modify experimental autoimmune thyroiditis (EAT) induced in mice immunized with mouse thyroglobulin and lipopolysaccharide. In this study, in an effort to elucidate the mechanisms by which GGT suppresses EAT, and autoimmunity in general, we investigated the in vivo effects of this drug on the Th1/Th2 lymphocyte balance, which is important for the induction or inhibition of autoreactivity. Naive SJL/J mice were treated orally for 5 days with GGT (80 mg/(kg day)). Spleen cells were obtained at various time points during the treatment period and were stimulated in vitro with concanavalin A. Interleukins IL-4, IL-10 and IL-12, transforming growth factor-beta (TGF-beta) and interferon-gamma (IFN-gamma) cytokine production was evaluated at the protein levels of the cytokines in the medium and mRNA expressions. A significant upregulation of IL-4, IL-10 and TGF-beta was observed following treatment with GGT, which peaked at day 5 (IL-10) or day 10 (IL-4). On the other hand, IL-12 and IFN-gamma production were either unchanged or decreased. It seems therefore that GGT induces in vivo a shift towards Th2 lymphocytes which may be one of the mechanisms of down-regulation of the autoimmune reactivity in EAT. Our observations indicate that down-regulation of TH1 cytokines (especially IL-12) and enhancement of Th2 cytokine production may play an important role in the control of T-cell-mediated autoimmunity. These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.     

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.     

Selective Th1 up-regulating activity of Withania somnifera aqueous extract in an experimental system using flow cytometry

Withania somnifera (Ashwagandha) is reported to be immunoprotective and immunoadjuvant. We studied its roots aqueous extract on T helper (Th) immunity using flow cytometry. This extract was standardized with six withanolides as marker compounds using HPLC. Once daily dose ranging from 25 to 400 mg/kg/p.o. was used to study effect on Th1: IFN-gamma, IL-2 and Th2: IL-4 cytokine modulation. We also studied effect on CD4 and CD8 in normal and immunesuppressed mice. The results indicate that extract at 100 mg/kg resulted significant selective up-regulation of Th1 response. Treatment with extract showed significant increase in CD4 and CD8 counts as compared to control and cyclopsorin A, with a faster recovery of CD4+ T cells in immunesuppressed animals. Under immunesuppressed conditions, potentiation of cellular and humoral immune responses of extract was comparable to levamisole. This study indicates the selective Th1 up-regulating effect of extract and suggests its use for selective Th1/Th2 modulation.     

雷公藤多甙对哮喘患者Th1、Th2细胞因子产生的影响

目的：了解雷公藤多甙对哮喘患者Th1,TH2细胞因子的影响，进一步探讨雷公藤多甙治疗哮喘的,机制。方法：选12例中，重度哮喘患者，口服雷公藤多甙每日40mg 或60mg治疗4周，治疗前后取外公藤甲素处理24h，收集培养上清，用酶联免疫吸附试验（ELISA）对患者血清及PBMC培养上清IL－2，IL－4，IL－5，γ干扰素（IFN－γ）水平进行检测。结果：（1）雷公藤多甙治疗后哮喘患者血清中IL－2，IL－4，IL－5水平较治疗前降低（P＜0．01），而IFN-γ治疗前后均在检测灵敏度（25pg/ml)以下。（2）雷公藤甲素处理后哮喘患者PBMC分泌IL－2，IL－4，IL－5减少（P＜0．01，P＜0．05），而IFN－γ也在检测灵敏度以下。结论：雷公藤多甙对哮喘患者Th2细胞因子的产生具有明显的抑制作用，是治疗哮喘的重要机制，但雷公藤多甙对Th1细胞因子的产生也有抑制作用，说明雷公藤抑制Th2,Th1 细胞因子产生的作用无特异性。     

蓝萼甲素对小鼠脾细胞Th1/Th2型5种细胞因子水平的影响

目的：探讨蓝萼甲素对小鼠脾细胞Th1/Th2型细胞因子水平的影响。方法：采用Cytometric Bead Array (CBA)法检测蓝萼甲素对小鼠脾细胞Th1/Th2型5种细胞因子(IL-2，IFN-γ，TNF-α，IL-4，IL-5)水平的影响。结果：蓝萼甲素对ConA诱导的Th1细胞来源的细胞因子(IL-2，IFN-γ，TNF-α) 和Th2细胞来源的细胞因子(IL-4，IL-5)的产生均有显著的抑制作用，且蓝萼甲素对Th1细胞来源的细胞因子的抑制作用要强于Th2细胞来源的；对未经ConA活化的小鼠脾细胞，蓝萼甲素对IL-2，IFN-γ和TNF-α的产生有抑制作用，而对IL-4，IL-5则有促进作用。结论：蓝萼甲素的免疫抑制作用可能与其对Th1/Th2型细胞因子的影响有关。     

A natural sweetener, Momordica grosvenori, attenuates the imbalance of cellular immune functions in alloxan-induced diabetic mice

Momordica grosvenori (MG), a traditional medicinal herb in China used as a substitute sugar for obese and diabetic patients, exhibited an enhancement of immunity. Heme oxygenase-1 (HO-1) is among the acute phase proteins that play an important role in the inflammatory process and have antiinflammatory activities with their antioxidant properties. The hypothesis that MG could modulate an imbalance of the cellular immune system and prevent the progression of diabetes mellitus, via induction of HO-1 protein expression was investigated. Diabetes was induced in male Balb/c mice by intraperitoneal injection of alloxan (200 mg/kg). The mice were randomly assigned to non-diabetic and diabetic groups, and further subdivided according to their treatments: control (distilled water), low dose MG (150 mg/kg) and high dose MG (300 mg/kg), which were administered by gavage in 24 h cycles over a 30 day period. MG administration effectively regulated the immune imbalance in diabetic mice, principally by upregulating the CD4(+) T lymphocyte subsets, and remodeling the intracellular cytokine profiles (reducing the expression of pro-inflammatory Th1 cytokines towards a beneficial Th2 pattern), ascribed to its induction and upregulation of HO-1. In conclusion, MG exhibited antidiabetic effects and may be used as a new natural drug for the treatment of diabetes.     

The anti-arthritic effects of Aconitum vilmorinianum, a folk herbal medicine in Southwestern China

Ethnopharmacological relevance

Aconiti Radix (AC) and Aconiti Kusnezoffii Radix (AK) are two traditional Chinese medicines commonly used to treat joint pain and arthritis. In Southwestern China, Huangcaowu (AV), the root of Aconitum vilmorinianum Kom., has long been used as a local substitute for these herbs for analgesia and anti-inflammation. However, its anti-arthritic effects have not been investigated.

Aim of study

To investigate the anti-arthritic effects of Huangcaowu (AV).

Materials and methods

Mono-arthritis in SD rats was induced by unilateral intra-articular injection of Freund's complete adjuvant. Physiological saline was injected in the contralateral knee. Seventy five percent ethanol extracts of AV (10 mg/kg/day and 100 mg/kg/day), AC (100 mg/kg/day) and AK (100 mg/kg/day) were administered to rats by oral gavage for 14 consecutive days (Day -6 to Day 7) while arthritis was induced at the seventh day (Day 0). The anti-arthritic effects of the herbs were assessed by measuring allodynia, swelling, hyperaemia and the vascular permeability of the knee joints.

Results

AV (10 mg/kg/day and 100 mg/kg/day), AC (100 mg/kg/day) and AK (100 mg/kg/day) suppressed joint allodynia. AV (10 mg/kg/day and 100 mg/kg/day) and AK (100 mg/kg/day) significantly reduced join swelling and hyperaemia while AC (100 mg/kg/day) did not. AV (100 mg/kg/day) attenuated vascular permeability while AC (100 mg/kg/day) and AK (100 mg/kg/day) showed no improvement.

Conclusions

Huangcaowu (AV) significantly improved allodynia, swelling, hyperaemia and vascular permeability in arthritic knee joints. It showed the highest anti-arthritic effects among the three tested Aconitum herbs.     

Experimental study on anti-inflammatory activity of a TCM recipe consisting of the supercritical fluid CO2 extract of Chrysanthemum indicum, Patchouli Oil and Zedoary Turmeric Oil in vivo

Ethnopharmacological relevance

Chrysanthemum indicum (Compositae) Linné, Pogostemon cablin (Blanco) Benth and Curcuma wenyujin (Zingiberaceae) Y. H. Chen et C. Ling are three of the extensively used herbal remedies among traditional Chinese medicines for the purpose of anti-inflammation. A traditional Chinese medicine (TCM) recipe named CPZ consisting extracts of the above three herbs, has shown noteworthy anti-influenza activity, which is closely related to its anti-inflammatory feature.

Aim of this study

To investigated the anti-inflammtory activity of CPZ in vivo for a further exploration of the recipe's anti-inflammatory properties.

Materials and methods

The anti-inflammatory property of CPZ on acute inflammation was evaluated by inflammatory models of dimethylbenzene (DMB)-induced ear vasodilatation and acetic acid-induced capillary permeability enhancement in mice, as well as the carrageenan-induced paw edema rat model, in which inflammation-related cytokine including prostaglandin E₂ (PGE₂), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and nitric oxide (NO) in the edematous paw tissue were determined by enzyme linked immunosorbent assay (ELISA). Moreover, effect of CPZ on chronic inflammation was observed through granuloma formation in rats subjected to cotton pellet implantation.

Results

CPZ (340, 170, and 85 mg/kg for mice, p.o.) not only decreased the DMB-induced ear vasodilatation but also attenuated capillary permeability under acetic acid challenge in mice. And the significant inhibition on carrageenan-induced paw edema was observed. Further more, the ELISA results showed that CPZ (170, 85, and 42.5 mg/kg for rats, p.o.) could up-regulate the level of IL-1β in the edema paw tissue of rats significantly while down-regulate that of PGE₂, but no apparent effect on TNF-α or NO was observed in the test. Besides, CPZ had a certain degree of restraining effect on the cotton pellet-induced granuloma formation in rats and the highest dose of 170 mg/kg even showed a significant suppression on it.

Conclusion

The above results indicated that CPZ possessed a potent anti-inflammatory activity, which is indicated to be closely associated with its regulation on IL-1β and PGE₂ thereby mediating the inflammatory response acting at an appropriate level.     

养阴益气活血方对干燥综合征NOD小鼠血清及颌下腺Th1/Th2免疫平衡的影响

目的 观察养阴益气活血方对干燥综合征（Sjogren′s Syndrome,SS）非肥胖型糖尿病（non-obese diabetic,NOD）小鼠血清及颌下腺干扰素－γ（interferon-γ, IFN-γ）、IL-2、IL-4、IL-10的表达及Th1/Th2免疫平衡的影响,探讨其作用机制。方法 32只NOD小鼠随机分为模型组、中药组［养阴益气活血方煎剂,0.4 mL（100 g/kg）］、西药组［羟氯喹0.4 mL（60 mg/kg）］、联合组［养阴益气活血方煎剂（50 g/kg）＋羟氯喹（60 mg/kg）0.4 mL］,每组8只,8只Balb/C小鼠作为正常对照（正常组）。实验进行8周后取血,摘取小鼠颌下腺组织,采用ELISA法检测小鼠血清IFN-γ、IL-2、IL-4、IL-10水平,SP法检测颌下腺组织IFN-γ、IL-2、IL-4、IL-10蛋白的表达。结果 模型组小鼠血清和颌下腺组织IFN-γ、IL-2、IL-4、IL-10水平明显高于正常组（P〈0.05）。中药组和联合组血清IFN-γ、IL-10水平均低于西药组（P〈0.05）。联合组颌下腺IFN-γ、IL-2表达低于西药组（P〈0.05）。模型组血清和颌下腺IFN-γ/IL-4比值均高于其他各组（P〈0.05）,且中药组和联合组IFN-γ/IL-4比值最接近正常组。结论 养阴益气活血方能降低SS NOD小鼠血清和颌下腺Th1、Th2相关细胞因子的水平,下调血清和颌下腺IFN-γ/IL-4比值。养阴益气活血方可能通过调节SS小鼠Th1/Th2免疫平衡达到治疗作用。     

Antinociceptive properties of bergenin

Bergenin (1) is a C-glucoside of 4-O-methylgallic acid with known antiarthritic activity attributed to modulation of cytokine production. The present study was undertaken to evaluate whether 1 has antinociceptive properties in models of inflammatory pain and to investigate its possible mechanisms of action. Pretreatment with 1 (12.5-100 mg/kg, ip) produced a dose-related inhibition of acetic acid-induced writhing in mice. Furthermore, treatment with 1 (50 and 100 mg/kg) inhibited both the early and late phases in a formalin test. In addition, 1 (50 and 100 mg/kg, ip) inhibited mechanical hyperalgesia, edema, and paw production of hyperalgesic cytokines (TNF-α and IL-1β) induced by complete Freund's adjuvant. However, the local production of IL-10, an anti-inflammatory cytokine, was not altered by 1 (100 mg/kg, ip). Treatment with 1 produced a similar profile of antinociception in wild-type and IL-10-deficient mice. Mice treated with 1 did not show any motor performance alterations or apparent systemic toxicity. The results presented herein demonstrate that bergenin has consistent antinociceptive and anti-inflammatory properties, acting by the inhibition of IL-1β and TNF-α production, and suggest its potential for the control of inflammatory pain.     

Effects of Catalpa ovata stem bark on atopic dermatitis-like skin lesions in NC/Nga mice

Ethnopharmacological relevance

The stem bark of Catalpa ovata has been used as a traditional herbal medicine for the treatment of various inflammatory diseases such as itching and scabies.

Aim of the study

In the present study, we investigated the anti-AD effects of Catalpa ovata stem bark on Dermatophagoides farinae-induced AD in a NC/Nga mouse AD model. We determined dermatitis score, histology, IgE, cytokines, and chemokines related to hypersensitive immune responses in AD. The mechanism of action was also investigated using HaCaT cells.

Materials and methods

We investigated the topical effects of Catalpa ovata stem bark on AD-like skin lesions in NC/Nga mice. Five category-experiments were performed, including assessment of dermatitis score; histological analysis of dorsal skin lesions; quantitative measurement of serum total IgE; quantitative measurement of cytokines (IL-1β, IL-4, IL-5, IL-6, IL-13, TNF-α) from dorsal tissue; and RT-PCR analysis of for TSLP and TARC mRNA expression in HaCaT cells.

Results

The clinical dermatitis score was significantly lower in Catalpa ovata extract (COE) groups than in the control group. Histological analysis showed that COE inhibited hypertrophy and hyperkeratosis of the epidermis, intracellular edema, and reduced the infiltration of inflammatory cells. COE significantly inhibited serum total IgE; Th2 cytokines IL-4, IL-5 and IL-13; pro-inflammatory cytokines IL-1β, IL6 and TNF-α; the Th2 chemokine TARC and the pro-Th2 cytokine TSLP.

Conclusion

These results demonstrate that Catalpa ovata stem bark may be a useful external medicine for treatment of AD. Further investigation is necessary to determine appropriate COE dosage and to evaluate the safety of this medicinal herb.