培土清心方对特应性皮炎患者血清n-6必需脂肪酸的影响

目的:通过检测特应性皮炎(AD)患者治疗前后n-6必需脂肪酸水平的变化,探讨培土清心方治疗AD的可能作用机制.方法:选取血清总IgE升高的患者采用清心培土方水煎剂口服,每日1剂,连续12周,用SCORAD指数进行病情严重度评估;检测治疗前后患者血清中亚油酸(LA)、γ-亚油酸(GLA)、二高亚油酸(DGLA)、花生四烯酸(AA)水平,以20例健康者为对照进行比较.结果:治疗前血清GLA明显低于正常对照组(P＜0.05);血清LA,DGLA,AA水平与对照组比较无差异;GLA/LA水平低于正常对照组(P＜0.05);血清GLA与SCORAD指数存在中度负相关(P＜0.05).治疗后SCORAD指数较治疗前明显下降,差异有统计学意义(P＜0.01),血清GLA、GLA/LA均较治疗前明显升高(P＜0.05).结论:IgE升高的AD患者血清GLA水平与病情严重程度相关,培土清心方治疗AD的作用机制可能与调节n-6去饱和酶作用有关.     

培土清心方对特应性皮炎患者血清总IgE、ECP影响的研究

目的：检测特应性皮炎（以下简称AD）患者血清中总IgE、ECP,以探讨二者在特应性皮炎发病机制中的作用;研究培土清心方对AD患者血清总IgE、ECP的影响,探求其治疗特应性皮炎可能的机制,以指导临床实践。方法：选择55例AD患者为研究对象,于治疗前及治疗后1个月、3个月分别记录SCORAD积分。并随机选取38名于治疗前及治疗后3个月时进行IgE、ECP的检测,另以15名健康志愿者为对照。结果：55例AD患者经培土清心方治疗3个月后SCORAD积分较治疗前有明显的下降。AD患者的治疗前血清IgE、ECP水平显著高于正常对照组（P〈0.01）,治疗后IgE、ECP水平明显降低（P〈0.01）。结论：AD患者血清总IgE、ECP水平明显升高,培土清心方治疗AD,可明显降低血清中总IgE、ECP水平,说明其治疗机制可能与免疫学机制相关。     

培土清心方治疗特应性皮炎的疗效观察及对血清NGF、SP影响的研究

目的:观察培土清心方治疗AD的疗效,以及对AD患者NGF、SP物质的影响。方法:选择55例AD患者为研究对象,于治疗前及治疗后1个月、3个月分别记录SCORAD积分。并在55名患者中随机选取38名于治疗前及治疗后3个月时进行血清学指标的检测,以15例健康志愿者为对照。结果:55例AD患者经培土清心方治疗3个月后SCO-RAD积分较治疗前有明显的下降。AD患者的治疗前血清NGF、SP水平显著高于正常对照组(P<0.01)。治疗后NGF、SP水平明显降低(P<0.01)。相关性分析表明:SCORAD积分与NGF、SP呈正相关(R=0.639,P<0.05);结论:(1)培土清心方治疗AD有确切疗效。(2)证实NGF及SP参与了AD的发病,支持AD发病机制中的神经源性机制。(3)AD患者治疗前血清NGF及SP水平与Rajka评分和SCORAD积分均存在正相关关系,提示NGF及SP与AD病情严重程度有关。(4)培土清心法治疗AD,可明显降低血清中NGF、SP水平,说明其治疗机制可能与神经源性机制有关。     

培土清心颗粒对豚鼠皮肤瘙痒模型的止痒效果研究

目的:观察评价培土清心颗粒对实验性瘙痒动物模型的止痒效果。方法:采用动物实验研究方法,观察培土清心颗粒对磷酸组胺所致豚鼠瘙痒模型、4-氨基吡啶所致小鼠瘙痒模型和右旋糖酐40诱发小鼠瘙痒模型的影响,观察培土清心颗粒的止痒作用。结果:培土清心颗粒可增加豚鼠涂抹磷酸组胺次数(P<0.01),升高豚鼠致痒阈值(P<0.01),减少4-氨基吡啶致瘙痒小鼠和右旋糖酐40诱发瘙痒小鼠的搔抓次数(P<0.01,P<0.05)。结论:培土清心颗粒可通过减少内源性和外源性组胺释放途径缓解皮肤瘙痒症状,对特应性皮炎具有良好的止痒效果。     

培土清心方联合耳穴压豆治疗脾虚湿蕴型特应性皮炎疗效观察

目的：观察培土清心方联合耳穴压豆治疗脾虚湿蕴型特应性皮炎的临床疗效。方法：将86例脾虚湿蕴型特应性皮炎患者以信封法分为两组各43例。对照组予培土清心方治疗,治疗组在对照组基础上予耳穴压豆治疗。治疗4周后,比较两组临床疗效。结果：治疗组总有效率为93.02%,高于对照组的74.42%(P﹤0.05)。治疗后,两组皮损面积及瘙痒和睡眠程度、DLQI、EASI及IGA评分均改善（P﹤0.05）,且治疗组优于对照组（P﹤0.05）;两组NRS评分及Ig E指标水平均降低（P﹤0.05）,且治疗组低于对照组（P﹤0.05）。结论：培土清心方联合耳穴压豆治疗脾虚湿蕴型特应性皮炎患者临床疗效较好,能降低患者瘙痒和睡眠影响程度,提高生活质量,促进皮损和瘙痒症的恢复,加快Ig E指标的复常。     

培土清心颗粒对小鼠特应性皮炎模型脾脏Th1/Th2细胞的影响

目的:观察培土清心颗粒对半抗原诱导C57BL/6小鼠特应性皮炎模型脾脏Th1/Th2细胞的影响。方法:18只雄性C57BL/6小鼠随机分为空白组、模型组、培土清心颗粒组,每组6只。模型组和培土清心颗粒组采用2,4-二硝基氟苯(DNFB)诱导C57BL/6小鼠建立特应性皮炎模型,从造模第1天开始,培土清心颗粒组小鼠按2.47 g/(kg·d)灌胃培土清心颗粒,空白组和模型组给予同等体积的蒸馏水灌胃,连续给药14天。观察小鼠皮损表现和皮肤组织病理切片情况,检测耳部肿胀度,使用流式细胞仪检测小鼠脾脏中Th1、Th2细胞的表达。结果:与空白组比较,模型组小鼠右耳厚度和Th2细胞显著升高,Th1/Th2显著降低,培土清心颗粒组小鼠右耳厚度明显升高,差异均有统计学意义(P0.01)。与模型组比较,培土清心颗粒组小鼠右耳厚度和Th2细胞显著降低,Th1/Th2显著升高,差异均有统计学意义(P0.05,P0.01)。空白组小鼠皮肤正常;模型组小鼠背部皮肤增厚、粗糙、红斑、脱屑、糜烂、结痂,皮肤组织病理表现为表皮角化不全、角化过度、海绵水肿、棘层增厚,真皮内大量淋巴细胞浸润;培土清心颗粒组小鼠背部皮肤轻度增厚、粗糙、淡红斑、少许脱屑、结痂,炎症程度较模型组减轻,皮肤组织病理表现为表皮轻度角化不全、角化过度、海绵水肿,棘层轻度增厚,真皮内少量淋巴细胞浸润。结论:C57BL/6小鼠特应性皮炎样模型存在以Th2细胞升高为主的Th1/Th2失衡,培土清心颗粒可降低该模型小鼠脾脏Th2细胞的表达,提示调节Th1/Th2失衡是培土清心颗粒抗炎作用的可能机制。     

基于网络药理学和蛋白模块探讨培土清心颗粒治疗特应性皮炎的作用机制

目的运用网络药理学和蛋白模块方法探讨培土清心颗粒治疗特应性皮炎(atopic dermatitis,AD)的作用机制。方法基于中药系统药理学数据库及分析平台(TCMSP)和BATMAN-TCM数据库获取培土清心颗粒化学成分及其对应靶点,通过GeneCards、OMIM、DisGeNET数据库筛选特应性皮炎疾病靶点。使用String数据库构建蛋白相互作用(PPI)网络;使用Cytoscape软件绘制培土清心颗粒潜在活性成分-靶点-疾病网络图,使用Network Analyzer工具对网络进行拓扑属性分析;以插件ClusterMaker对靶点进行模块化分解,并使用DAVID数据库进行基因百科全书(KEGG)通路富集分析。结果得到培土清心颗粒治疗特应性皮炎活性成分185个,靶点125个,重要活性成分包括檞皮素、木犀草素、山奈酚、汉黄芩素等;重要靶点包括PTGS2、CXCL8、IL-10、CCL2等;PPI网络模块化分析结果显示,125个靶点主要分为3个模块,KEGG通路富集方面,模块一内靶点KEGG通路主要与特应性皮炎免疫系统、炎症反应相关;模块二内靶点KEGG通路主要与细胞增殖、分化、凋亡相关;模块三内靶点KEGG通路与代谢密切相关。结论培土清心颗粒可能通过抗炎、调节免疫、调控细胞凋亡以及调节氨基酸代谢以达到治疗特应性皮炎的目的。     

补肾解郁清心方结合针刺对围绝经期抑郁症患者ACT-INH-FS通路的影响

目的探讨补肾解郁清心方结合针刺治疗围绝经期抑郁症患者疗效及对激活素(ACT)-抑制素(INH)-卵泡抑素(FS)(ACT-INH-FS)通路的影响。方法将60例围绝经期抑郁症患者随机分成中药组和针药组各30例,中药组给予补肾解郁清心方口服,针药组给予补肾解郁清心方口服+针刺治疗,2组疗程均为12周。分别在治疗前及治疗12周末对2组患者进行HAMD评分和改良Kupperman量表评分,并检测血清性激素[卵泡刺激素(FSH)、雌二醇(E_2)及黄体生成素(LH)]、ACT、INH、FS水平。结果 2组治疗后HAMD评分、改良Kupperman量表评分均明显降低(P均<0.05),且针药组均明显低于中药组(P均<0.05)。2组治疗后FSH、LH、FS、ACTA水平均明显降低(P均<0.05),针药组治疗后E_2、INHB水平均明显升高(P均<0.05),针药组治疗后FSH、LH、E_2、FS、ACTA、INHB改善情况均明显优于中药组(P均<0.05)。针药组有效率明显高于中药组(P<0.05)。结论补肾解郁清心方单用和补肾解郁清心方联合针刺治疗均能缓解患者围绝经期抑郁症状,降低血清FSH、LH、FS、ACTA水平,但针药联合治疗疗效更好,且能提高血清E_2、INHB水平,对ACT-INH-FS通路影响更明显,可以更好地发挥抗抑郁作用。     

姜黄素对人胰腺癌细胞增殖抑制作用及对Wnt信号通路的影响

目的:探究姜黄素对人胰腺癌SW1990细胞增殖抑制作用及对Wnt信号通路的影响。方法:低/中/高剂量观察组人胰腺癌SW1990细胞分别以终浓度为20、50、100μmol/L姜黄素处理,100μL/孔,对照组细胞则以等体积生理盐水处理,分别干预24、48、72 h。以噻唑蓝(MTT)法及Annexin-V/FITC双染法分别检测人胰腺癌细胞SW1990增殖及凋亡情况,反转录聚合酶链式反应(RT-PCR)法及细胞爬片免疫细胞化学技术检测人胰腺癌细胞SW1990β-链蛋白(β-Catenin)mRNA及β-Catenin蛋白表达情况。结果:干预72 h后低/中/高剂量观察组人胰腺癌SW1990细胞增殖抑制率(%)分别为(4.59±1.32)、(40.01±3.87)、(60.98±5.97);对照组及低/中/高剂量观察组人胰腺癌SW1990细胞凋亡率(%)(8.14±1.25)、(13.04±1.03)、(27.15±1.62)、(59.21±1.46);β-Catenin蛋白免疫细胞化学(ICC)评分(分)分别为(5.78±0.56)、(4.42±0.61)、(2.57±0.58)、(1.38±0.63)。观察组凋亡率显著高于对照组,β-Catenin mRNA相对表达量及β-Catenin蛋白ICC评分显著低于对照组(P0.05),且增殖抑制率、凋亡率随姜黄素作用浓度增加逐渐升高(P0.05),增殖抑制率亦随干预时间延长逐渐升高(P0.05);β-Catenin mRNA相对表达量及β-Catenin蛋白ICC评分随姜黄素作用浓度增加逐渐降低(P0.05)。结论:姜黄素可有效下调人胰腺癌细胞SW1990β-Catenin基因表达,抑制Wnt信号通路活化,具有显著的增殖抑制作用,且具有明显的时间-剂量依赖性。     

基于ERK/MAPK信号通路探讨青藤碱对SW480结肠癌细胞活性抑制作用的研究

目的:探讨青藤碱对SW480结肠癌细胞活性抑制作用及对细胞外调节蛋白激酶(extracellu-lar regulated protein kinase,ERK)/丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)信号通路的影响.方法:选用SW480人结肠癌细胞株,设对照组...     

Involvement of p38 MAPK signaling pathway in the anti-melanogenic effect of San-bai-tang, a Chinese herbal formula, in B16 cells

Aim of the study

San-bai-tang (SBT), a Chinese herbal formula, is traditionally used as a skin whitener in China. In our previous screening assays, SBT was identified as an effective tyrosinase inhibitor. In this study, we aim to investigate the anti-melanogenic effect and mechanisms of SBT in B16 cells.

Materials and methods

Cell viability was examined by the MTT assay. Cellular tyrosinase activity and melanin content were determined using spectrophotographic methods. Protein expression was analyzed by immunoblotting.

Results

SBT inhibited tyrosinase activity with an IC₅₀ of 215.6 ± 10.3 μg/ml, and decreased cellular melanin content with an IC₅₀ of 254.8 ± 14.5 μg/ml at 48 h. MTT assay demonstrated that 48-h SBT (50-400 μg/ml) treatment did not show obvious cytotoxicity. Immunoblot analysis showed that SBT (100, 200 or 400 μg/ml) treatment for 48 h down-regulated the expression levels of phosphorylated-p38, MITF, tyrosinase, TRP-1 and TRP-2 in a dose-dependent manner.

Conclusions

SBT inhibited melanogenesis in B16 cells, and suppression of p38 MAPK signaling pathway contributed to the anti-melanogenic effect of SBT by down-regulating the expression of MITF and melanogenic enzymes. These novel findings demonstrated the anti-melanogenic effect and mechanisms of SBT, and provide pharmacological basis for the traditional use of SBT.     

中药QHF复方抑制乳腺癌MCF-7细胞增殖、迁移、侵袭的机制研究

摘要：目的 研究中药QHF复方对乳腺癌MCF-7细胞增殖、迁移、侵袭的影响及其可能的作用机制,为QHF的临床应用提供理论依据。方法 体外培养人乳腺癌MCF-7细胞,以对数生长期的细胞为研究对象,予以不同浓度QHF复方及其他药物,CCK8法检测QHF复方对乳腺癌MCF-7细胞生长增殖的影响。细胞划痕实验和Transwell实验检测QHF复方对乳腺癌MCF-7细胞迁移、侵袭的影响。Western blot检测QHF复方对乳腺癌MCF-7细胞HGF/c-Met及其下游PI3K/Akt、MAPK/ERK信号通路的影响。结果 CCK8实验结果显示：中药QHF复方可不同程度抑制乳腺癌MCF-7细胞增殖且这种抑制作用可显著拮抗HGF对乳腺癌MCF-7细胞增殖的促进作用。细胞划痕实验和Transwell实验结果显示：中药QHF复方能明显抑制MCF-7细胞迁移、侵袭且这种抑制作用可拮抗HGF 对MCF-7细胞迁移、侵袭的促进作用。Western blot实验结果显示：中药QHF复方可明显抑制乳腺癌MCF-7细胞HGF、p-c-Met、p-Akt、p-ERK、VEGF、MMP-2、MMP-9的表达。结论 中药QHF复方抑制乳腺癌MCF-7细胞的增殖、迁移和侵袭的机制可能与抑制乳腺癌MCF-7细胞异常活化的HGF/c-Met及其下游PI3K/Akt、MAPK/ERK信号通路有关。     

Clinical evaluation of the Chinese herbal medicine formula STA-1 in the treatment of allergic asthma

Although some formulae of traditional Chinese medicines (TCM) have been used for antiasthma treatment, few of them have had sufficient discussion on their efficacy, safety and mechanisms. In this study, the availability of the TCM formula STA-1 for the treatment of allergic asthma was investigated by conducting a double-blind, placebo-controlled and randomized trial. One hundred and twenty patients between the ages of 5 to 20 years with mild-to-moderate asthma were included. These patients were treated with either STA-1 or placebo in a dose of 80 g/kg/day and were administered twice daily for 6 months. The main outcome measures were a daily diary record of symptoms, supplementary bronchodilator and glucocorticoid treatment, changes of pulmonary function (forced expiratory volume in 1 s), changes of total and Dermatophagoides pteronyssinus (DP)-specific IgE and side effects. The results showed a statistically significant reduction of symptom scores, systemic steroid dose, total IgE and specific IgE in the STA-1 group. Furthermore, STA-1 also improved the pulmonary lung function FEV(1) compared with the placebo group and only minimal side effects were shown. These results suggested that STA-1 is available for the treatment of mild-to-moderate chronic asthma.     

截断逆挽方含药血清对D-氨基半乳糖诱导的人正常肝细胞LO2细胞增殖影响

目的研究截断逆挽方含药血清通过调控E2F1介导的细胞增殖途径减轻D-氨基半乳糖(D-Galactosamine,D-GlaN)诱导的人正常肝细胞LO2的损伤。方法体外培养LO2细胞,以D-GlaN进行造模,将细胞分为正常组、模型组、2.5%截断逆挽方含药血清组(2.5%JDNWF)、5%截断逆挽方含药血清组(5%JDNWF)、10%截断逆挽方含药血清组(10%JDNWF)。采用Cell Counting Kit-8(CCK8)法检测细胞活力;Real-time PCR检测转录因子E2F1、细胞周期蛋白E(cyclin E)、细胞周期蛋白A(cyclin A)、细胞周期蛋白依赖性激酶2(cyclin-dependent kinase 2,CDK2)、细胞分裂周期因子25A(cell division cyclin 25A,CDC25A)mRNA的表达;流式细胞术检测细胞周期;Western Blot检测分化调控因子DP1蛋白表达。结果截断逆挽方含药血清能显著促进LO2细胞进入S期(P<0.01),上调E2F1、cyclin E、cyclin A、CDK2、CDC25A mRNA的表达(P<0.01),并且能增加DP1蛋白的表达(P<0.05,P<0.01)。结论截断逆挽方减轻肝细胞损伤的作用机制可能与其促进肝细胞增殖有关。     

归术益坤方对多囊卵巢综合征模型大鼠卵巢组织及SIRT1-FOXO信号通路相关蛋白表达的影响

目的 探讨归术益坤方对多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠卵巢功能的影响及其机制.方法 SPF级雌性SD大鼠40只,随机分为空白组、模型组、中药高剂量组、中药低剂量组、阳性对照组,每组8只.除空白组外,每组均高脂饲料喂养联合丙酸睾酮注射液肌注,建立PCOS大鼠模型.造模2周...     

Xiaoyao San,a Chinese herbal formula,ameliorates depression-like behavior in mice through the AdipoR1/AMPK/ACC pathway in hypothalamus

Objective:Depression and metabolic disorders have overlapping psychosocial and pathophysiological causes.Current research is focused on the possible role of adiponectin in regulating common biological mechanisms,Xiaoyao San(XYS),a classic Chinese medicine compound,has been widely used in the treatment of depression and can alleviate metabolic disorders such as lipid or glucose metabolism disorders.However,the ability of XYS to ameliorate depression-like behavior as well as metabolic dysfunction ...     

A traditional Chinese medicine formula extracts stimulate proliferation and inhibit mineralization of human mesenchymal stem cells in vitro

Aim of the study

To investigate the effects of a traditional Chinese medicine (TCM) formula extract, named as ZD-I, on the proliferation and osteogenic differentiation of human mesenchymal stem cells (hMSCs) in vitro.

Materials and methods

When hMSCs cultivated in the basal medium with ZD-I, cell viability was assessed by MTT assay and cellular proliferation was assessed by SYBR green I assay. The effects of ZD-I on osteogenic differentiation of hMSCs were assessed by alkaline phosphatase (ALP) activity, mineralization assay and real-time RT-PCR.

Results

ZD-I (0.78–100 μg/ml) was non-cytotoxic. The 50% inhibitory concentration (IC50) of hMSCs was 200 μg/ml. ZD-I (0.78–50 μg/ml) stimulated the proliferation of hMSCs. ZD-I did not change ALP activity of hMSCs cultivated in osteogenic medium in the early stage (4 and 7 days), but ZD-I inhibited the mineralization of hMSCs through down-regulation of several osteogenic markers (e.g. osteocalcin, bone morphogenetic protein 2 and osteopontin) in the late stage.

Conclusions

ZD-I stimulate cellular proliferation and decrease the bone mineral deposition of hMSCs. These results suggest ZD-I may play an important therapeutic role in osteoarthritic patients by improving proliferative capacity of hMSCs and inhibiting the mineralization of hMSCs.     

Biochemical mechanism of Wu-Zi-Yan-Zong-Wan,a traditional Chinese herbal formula,against alcohol-induced oxidative damage in CYP2E1 cDNA-transfected HepG2 (E47) cells

Aim of the study

Wu-Zi-Yan-Zong-Wan (WZ) is a traditional Chinese herbal formula which is commonly used for treating patients with “Yang deficiency”. In the present study, the effect of WZ on ethanol-induced toxicity in CYP2E1 cDNA-transfected HepG2 (E47) cells was investigated.

Materials and Methods

WZ extract was obtained by extracting the herbal powder with 50% ethanol (v/v, in water) and the effect of the extract on ethanol-induced toxicity was investigated in cultured cells.

Results

The treatment with WZ extract (12.5–200 μg/mL) for 24 h dose-dependently protected against ethanol-induced toxicity in E47 cells, as evidenced by the enhanced cell viability and decreased extent of lactate dehydrogeanse leakage. The cytoprotection against ethanol-induced toxicity was associated with decreases in the extents of reactive oxygen species production and lipid peroxidation, as well as increases in mitochondrial reduced glutathione and membrane potential. In addition, WZ extract treatment also suppressed the formation of DNA fragments in ethanol-intoxicated E47 cells.

Conclusions

WZ extract was found to protect against the ethanol-induced toxicity in E47 cells, possibly by virtues of its antioxidant activity.