中药熏蒸在治疗大鼠佐剂性关节炎中TH1/TH2的变化

目的:检测大鼠血液中炎性细胞因子IL-4I、L-10I、FN-γ、TNF-α,以阐明中药熏蒸对大鼠佐剂性关节炎的疗效机理。方法:建立大鼠AA模型,将大鼠随机分为正常组、模型组、蒸馏水熏蒸组、低浓度中药熏蒸组、高浓度中药熏蒸组、西药组。正常组与模型组大鼠不治疗,蒸馏水熏蒸组、低浓度中药熏蒸组、高浓度中药熏蒸组大鼠分别蒸馏水、低浓度中药熏蒸、高浓度中药熏蒸,西药组采用扶他林喷雾剂治疗,隔天测一次各组大鼠体重及双后肢足肿。一疗程(10天)后,比较各组大鼠血清中细胞因子IL-4I、L-10I、FN-γ、TNF-α的含量变化。结果:中药熏蒸可上调大鼠血清中细胞因子IL-4I、L-10水平,下降IFN-γ、TNF-α水平,且存在一定的量效关系。结论:中药熏蒸疗法的抗炎作用与调节TH1/TH2平衡有关。     

Morin promotes the production of Th2 cytokine by modulating bone marrow-derived dendritic cells

Our previous studies had reported that morin decreased the interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) production in lipopolysaccharide (LPS)-activated macrophages, suggesting that morin may promote helper T type 2 (Th2) response in vivo. Dendritic cells (DCs) are the most potent antigen presenting cells and known to play a major role in the differentiation of helper T type 1 (Th1) and Th2 responses. This study aimed to reveal whether morin is able to control the Th differentiation through modulating the maturation and functions of DCs. Bone marrow-derived dendritic cells (BM-DCs) were incubated with various concentrations of morin and their characteristics were studied. The results indicated that morin significantly affects the phenotype and cytokine expression of BM-DCs. Morin reduced the production of IL-12 and TNF-alpha in BM-DCs, in response to LPS stimulation. In addition, the proliferative response of stimulated alloreactive T cells was significantly decreased by morin in BM-DCs. Furthermore, allogeneic T cells secreted higher IL-4 and lower IFN-gamma in response to morin in BM-DCs. In conclusion, these results suggested that morin favors Th2 cell differentiation through modulating the maturation and function of BM-DCs.     

Traditional Korean medicine (SCRT) modulate Th1/Th2 specific cytokine production in mice CD4+ T cell

Traditional Korea medicine, So-Cheong-Ryong-Tang (SCRT) also called as Xiao-qing-long-tang or Sho-seiru-to, contains eight species of medicinal plants and has been used for treating allergic diseases, such as allergic rhinitis and asthma, for hundreds of years in Asian countries. CD4+ T cells were highly purified by using magnetic bead from splenocytes of BALB/c mice. SCRT treatment slightly decreased the expression of cell surface protein CD69 on CD4+ T cell in the flow cytometry analysis. In RT-PCR analysis, SCRT increases the expression of IL-2 and IL2R-alpha mRNA, and decreases the expression of IL-4 mRNA, which is an important cytokine of Th2 cell development. On the other hand, SCRT treatment increases IFN-gamma expression, which is one of the key cytokines for Th1 cell development. Present study implies that SCRT can correct Th2 dominant condition directly affecting to the CD4+ T cell without significantly depressing general T cell activities. These results also suggest that the effect on CD4+ T cell may be the one of key pharmacological effect point for treating IgE medicated allergic asthma by SCRT.     

Matrine attenuates allergic airway inflammation and eosinophil infiltration by suppressing eotaxin and Th2 cytokine production in asthmatic mice

Ethnopharmacological relevance

Matrine has been isolated from Sophora flavescens, and found to show anti-inflammatory effects in macrophages and anti-cachectic effects in hepatomas. The present study investigated whether matrine suppressed eosinophil infiltration and airway hyperresponsiveness (AHR) in mice, and decreased the inflammatory response of tracheal epithelial cells.

Materials and methods

BALB/c mice were sensitized and challenged with ovalbumin to induce allergic asthma in mice. These asthmatic mice were given various doses of matrine by intraperitoneal injection. Additionally, activated human tracheal epithelial cells (BEAS-2B cells) were treated with matrine, and evaluated for levels of proinflammatory cytokines and chemokines.

Results

We found that matrine significantly decreased AHR, and suppressed goblet cell hyperplasia, eosinophil infiltration, and inflammatory response in the lung tissue of asthmatic mice. Matrine also reduced the levels of Th2 cytokines and chemokines in bronchoalveolar lavage fluid, and suppressed OVA–IgE production in serum. Furthermore, matrine treatment of activated BEAS-2B cells decreased production of proinflammatory cytokines and eotaxins, as well as suppressed ICAM-1 expression and thus adhesion of eosinophils to inflammatory BEAS-2B cells in vitro.

Conclusions

Our findings suggest that matrine can improve allergic asthma in mice, and therefore has potential therapeutic potential in humans.     

心理应激诱发实验性2型糖尿病大鼠的行为学改变及中药干预

[目的]从宏观行为学角度证实情志疾病基本病机—气机失调的客观存在及复方中药的干预作用。[方法]采用多项心理应激结合小剂量链脲佐菌素腹腔注射的复合方式，模拟心理应激诱发实验性2型糖尿病的大鼠模型，借鉴经典的旷场实验方法评定心理应激对2型糖尿病大鼠行为的影响，同时观察了具有理气、降逆散结作用的复方中药心身5号的调节作用。[结果]心理应激可导致实验性2型糖尿病大鼠的行为异常，心身5号可调节异常的行为改变。[结论]心理应激诱发实验性2型糖尿病大鼠行为异常是气机失调的外在宏观表现，反映情志疾病的共同病机气机失调的客观存在，及具有理气降逆散结作用的复方中药心身5号具有明显的调节效应。     

大鼠模型在更年期综合征中西医实验研究中的应用概述

归纳和总结了近年大鼠模型在更年期综合征中西医实验研究中的应用。本文概述了该病的大鼠模型研制现状、模型大鼠应用于发病机制和药物作用机理的研究进展,并提出了实验研究中存在的问题。大鼠模型在该病的研究中,发病机理研究主要集中在血管舒缩状态、精神神经症状、心血管疾病、骨矿含量改变及骨质疏松等方面;西药作用机理的研究主要集中在对各种人工激素和化学合成药物对更年期综合征临床表现的作用及作用机理研究;中药的研究主要集中在对中药单一成分、中药复方、中药有效部位群、中成药、中药制作工艺、量效关系及经方的研究;针灸治疗更年期综合征的基础研究主要针对其"治未病"的预防保健作用和对临床症状的治疗作用机理,以及取穴和穴位埋线治疗作用等。     

Effect of ninjin-youei-to on Th1/Th2 type cytokine production in different mouse strains

Ninjin-youei-to (NYT; ren-shen-yang-rong-tang in Chinese) is a traditional herbal formula, which is widely used in Japan, Korea and China to modulate physiological immunity. The effects of oral administration of NYT on cytokine production from splenocytes were investigated in both C57BL/6 and BALB/c mice in which Th1 and Th2 were dominant, respectively. Splenocytes from C57BL/6 and BALB/c mice, which took NYT orally forfour weeks, were cultured with anti-mouse CD3 mAb, and the supernatant was examined for cytokine production using enzyme-linked immunosorbent assay (ELISA). Administration of NYT to C57BL/6 mice, increased the production of interleukin-4 (IL-4) significantly, and slightly decreased interferon-y (IFN-gamma) production from splenocytes. In contrast, the same treatment significantly increased IFN-gamma secretion from splenocytes of BALB/c mice. No remarkable changes of IL-12 production from splenocytes were observed in either strain of mice. These results suggest that oral administration of NYT ameliorates the excessive inclination of Th1 and Th2 type cytokine production, and NYT may provide a beneficial effects for the treatment of diseases caused by a skewed Th1-Th2 balance in the immune system.     

定喘汤联合西医常规治疗对过敏性支气管哮喘患者外周血单个核细胞Th1/Th2及黏膜免疫功能的影响

目的观察定喘汤联合西医常规治疗对过敏性支气管哮喘患者外周血单个核细胞Th1/Th2及黏膜免疫功能的影响。方法将80例过敏性支气管哮喘患者按随机数字表法分为2组,对照组40例予西医常规治疗,治疗组40例在对照组治疗基础上加用定喘汤治疗。2组均治疗4周后统计临床疗效,并观察2组治疗前后喘息、咳嗽、咯痰及哮鸣音症状积分,第1 s用力呼气容积(FEV_1)、FEV_1/用力肺活量(FVC)、第1 s用力呼气容积占预计值百分比(FEV_1%),外周血单个核细胞辅助性T细胞1百分率(Th1)、外周血单个核细胞辅助性T细胞2百分率(Th2)、Th1/Th2,气道基质金属蛋白酶-9(MMP-9)评分、嗜酸性粒细胞计数,以及不良反应发生情况。结果 2组治疗后喘息、咳嗽、咯痰及哮鸣音中医症状积分均改善(P0.05),且治疗组改善优于对照组(P0.05)。2组治疗后FEV_1、FEV_1/FVC、FEV_1%、Th1百分率、Th2百分率、Th1/Th2、气道MMP-9评分及嗜酸性粒细胞计数均改善(P0.05),且治疗组改善优于对照组(P0.05)。治疗组总有效率90.00%,对照组总有效率60.00%,2组比较差异有统计学意义(P0.05),治疗组疗效优于对照组。2组间不良反应发生率比较差异无统计学意义(P0.05)。结论定喘汤联合西医常规治疗过敏性支气管哮喘疗效确切,并能改善临床症状、肺功能、外周血单个核细胞TH1/TH2及黏膜免疫功能。     

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.     

膈下逐瘀汤对肝纤维化大鼠组织形态学的影响

目的:从形态学角度探讨膈下逐瘀汤对肝纤维化大鼠胶原代谢的干预作用。方法:采用ALA- kokkol腹腔注射DMN的方法制作肝纤维化动物模型。结果:光镜和电镜下膈下逐瘀汤及其拆方各组炎性细胞浸润,纤维组织增长和其它各组对比有明显差异。结论:膈下逐瘀汤对肝纤维化大鼠胶原纤维增生、毛细血管化等具有明显减轻作用。     

狼疮方2号对狼疮模型血清中几种细胞因子的影响

 目的研究中药复方狼疮方2号(LCF-2)对BXSB狼疮模型鼠血清中细胞因子的作用及对脾指数、胸腺指数的影响,探讨其对SLE的作用机制。方法 将BXSB狼疮模型鼠随机分为空白对照组、中药组、西药组、中西药合用组。重量法测定小鼠脾指数及胸腺指数;双抗夹心酶联免疫吸附法(ELISA)测定小鼠血清中IL-2,IL-6,TNF-a的含量。结果 LCF-2组能够升高已降低的胸腺指数,与空白对照组比差异有显著性,对已升高的脾指数作用不明显,而中西药组可以降低升高的脾指数(P<0.01),升高降低的胸腺指数(P<0.05);中药组能显著抑制TNF-a(P<0.05),上调IL-2水平(P<0.01),对IL-6的降低作用无统计学意义;中西药合用药组能抑制TNF-a(P<0.01),上调IL-2水平(P<0.05),抑制IL-6(P<0.05)。结论 中药复方LCF-2与西药泼尼松均对BXSB狼疮小鼠有明显的治疗作用,但两者的作用机制不同。中西药合用组疗效优于中药、西药组。     

从信号通路角度分析中药治疗儿童白血病的研究进展

白血病是儿童最常见的恶性肿瘤,也是通过化疗可以治疗的疾病,但感染和化疗药物的毒副作用是导致儿童白血病患者死亡的原因,因此,临床中常采用中药联合化疗药物的方式治疗儿童白血病,尽管中药在治疗白血病方面具有广泛的临床经验,但作用机制并不清楚。目前,靶向药物由于具有选择性高和毒副作用小的优势是研究的热点,在白血病领域的靶向药物主要是针对BCR/ABL,TARA和FLT3蛋白,它们的异常引起下游PI3K/AKT,JNK/STAT,Ras/ERK等信号通路的变化。近年来发现治疗白血病的中药的作用靶点或作用通路逐步清晰,该文总结具有调节信号通路作用的中药复方或单体成分,为中药联合化疗药物或靶向药物治疗儿童白血病提供依据。     

Inhibition of proliferation and induction of apoptosis by tetrandrine in HepG2 cells

Tetrandrine is a bisbenzylisoquinoline alkaloid derived from the root of Stephania tetrandra S. Moore, which has been reported to elicit in vitro cytotoxic effects on HeLa cells, and in vivo suppressive effects on mouse ascites tumors. In the present study, we examined the antiproliferative and apoptosis-inducing activity of tetrandrine in HepG2 cells, a human hepatoma cell line. Tetrandrine showed potent cytotoxic activity in HepG2 cells (IC(50)=9.0+/-1.0 micro M) following incubation for 48 h. Dose-dependent induction of apoptosis was observed by agarose gel electrophoresis and flow cytometric analysis. Treatment of HepG2 cells with tetrandrine resulted in the activation of caspase-3 protease, and subsequent proteolytic cleavage of poly(ADP-ribose) polymerase. These results suggest that tetrandrine is potentially useful as a chemotherapeutic/chemopreventive agent in hepatocellular carcinoma.     

Protective effects of Jiayan Kangtai granules on autoimmune thyroiditis in a rat model by modulating Th17/Treg cell balance

OBJECTIVE: To investigate the protective effects of Jiayan Kangtai(JYKT) granules, consisting of 9 Chinese herbs, in a rat model of autoimmune thyroiditis(AIT), and the possible underlying mechanism.METHODS: Female Lewis rats(6-8 weeks) were randomly apportioned to 5 groups of 10, including a normal control. AIT was induced in the untreated AIT-model group, and rats treated subsequently with daily low, medium, or high dose JYKT granules. After 12 weeks, plasma levels of thyroid autoantibodies and morphological changes in the thyroid were detected by enzyme-linked immunosorbent assay and histological examination, respectively. The presence of interleukin(IL)-6,IL23 p19, and IL-2 in thyroid tissue was assessed by immunohistochemical staining. The percentages of T helper(Th)17 cells and regulatory T cells(Tregs) in the peripheral blood were analyzed by flow cytometry. Relevant levels of cytokines and proteins were examined via bead-based multiplex flow cytometry and ELISA, respectively. Expressions of genes and proteins regulated by Th17 cells and Tregs were shown by real-time PCR and Western blot.RESULTS: Compared to the control, AIT-model rats had higher plasma concentrations of thyroid autoantibodies. The high-dose JYKT rats showed significantly lower levels of thyroid autoantibodies compared with the AIT model group. Rats in the AIT-JYKT groups also had fewer thyroid lesions and less lymphocytic infiltration, a lower percentage of Th17 cells, and a higher percentage of Tregs, compared with the AIT-model. Rats given high-dose JYKT had a significantly lower Th17/Treg ratio compared with the AIT model. Differences in plasma cytokine concentrations and relevant gene and protein expressions in the spleens of JYKT-treated rats and the AIT group suggested an association between JYKT treatment and lower Th17 cell percentage and higher Treg activity.CONCLUSION: JYKT treatment appeared to be protective against AIT in rats, possibly via the regulation of the Th17 cell/Treg imbalance in AIT.     

原儿茶酸对帕金森病模型小鼠中脑和纹状体D2DR、iNOS和TH表达的影响

目的考察原儿茶酸对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)致帕金森病(PD)小鼠中脑和纹状体多巴胺D2受体(D2DR)、诱导型一氧化氮合酶(i NOS)和酪氨酸羟化酶(TH)表达的影响。方法选用C57BL褐鼠为实验动物,连续7 d ip MPTP(25 mg/kg)制备PD动物模型。造模前后分别ip给予原儿茶酸(10 mg/kg)或阳性对照药美多芭(125 mg/kg)进行预防和治疗。应用Western blotting的方法,观察原儿茶酸对PD模型小鼠中脑和纹状体内D2DR、i NOS和TH表达的影响。结果 PD模型小鼠脑组织内i NOS表达升高,D2DR和TH表达下降,而原儿茶酸能够显著降低PD模型小鼠脑中i NOS的表达,提高D2DR和TH的表达量。结论原儿茶酸对MPTP诱导的PD模型小鼠有保护作用,可能通过提高PD模型小鼠中脑和纹状体内D2DR和TH的表达,降低i NOS的表达发挥作用。     

Preventive effect of Ophiopogon japonicus polysaccharides on an autoallergic mouse model for Sjogren's syndrome by regulating the Th1/Th2 cytokine imbalance

AIM OF STUDY: Sjogren's syndrome (SS) is an autoimmune disorder characterized by lymphocytic infiltration of salivary and lacrimal glands leading to xerostomia and keratoconjunctivitis sicca. Evidence has accumulated suggesting that a Th1/Th2 cytokine imbalance has a role in the pathogenesis of SS. Currently, only palliative treatment is available. Ophiopogon japonicus, a common Chinese herbal, has been used to treat sicca-associated disorders in traditional Chinese medicine for centuries. MATERIALS AND METHOD: In this study, we constructed an autoallergic mouse model for SS by immunizing C57BL/6 mouse with submandibular gland (SMG) autoantigen. At the same time, Ophiopogon japonicus polysaccharides (OJP) was administered and hydroxychloroquine was served as positive control. During the 4 weeks' experiment, salivary flow rates were determined every week, body weight, food and water intake were measured every 2 days. After death, serum were collected for IFN-gamma and IL-4 ELISA analysis and the IFN-gamma/IL-4 was calculated, SMG and spleen were harvested for organ index calculation, and part of SMG was examined for histological changes. RESULTS: Results showed that immunization with SMG autoantigen induced decreased salivary flow and body weight, increased water intake, SMG index, spleen index, IFN-gamma level and IFN-gamma/IL-4 ratio compared with the normal group. However, administration of the OJP could improve these data and the pathological changes of SMG with respect to the model mice, especially the high-dose of OJP group. CONCLUSIONS: Thus, this study provided a basis for the use of Ophiopogon japonicus in SS.     

HPLC法测定清补平衡膏方中没食子酸、5-HMF和芍药苷的含量

目的:建立同时测定清补平衡膏方中没食子酸、5-HMF和芍药苷含量的高效液相色谱法。方法:采用Hibar ODS柱(4.6 mm×250 mm,5μm),以乙腈-0.1%甲酸溶液为流动相梯度洗脱,流速为1.0 m L·min-1,检测波长为230 nm,柱温35℃,进样量10μL。结果:没食子酸在13.45~672.75μg·m L-1(r=0.999 4)范围内,5-HMF在10.76~537.9μg·m L-1(r=0.999 8)范围内,芍药苷在17.69~884.61μg·m L-1(r=0.999 8)范围内,对照品进样浓度和色谱峰峰面积线性关系良好,加样回收率分别为99.80%、96.94%和99.46%,RSD值分别为2.91%、2.93%和2.97%。结论:建立的高效液相色谱法简单易行,结果准确可靠,可有效地用于清补平衡膏方的质量控制。