熵权法结合星点设计-效应面法优化祛湿清肺方提取工艺及体外抗炎活性评价

目的:优化祛湿清肺方提取工艺,并对祛湿清肺方提取液进行体外抗炎活性评价。方法:以绿原酸、虎杖苷、黄芩苷、汉黄芩苷、黄芩素、芦荟大黄素、汉黄芩素、大黄素成分含量及得膏率为指标,加水量及提取时间为考察因素,采用熵权法结合星点设计-效应面法对祛湿清肺方提取工艺进行优化。以脂多糖(LPS)诱导大鼠腹腔巨噬细胞(RAW_264.7)为炎症模型,酶联免疫吸附法测定白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、一氧化氮(NO)的含量,蛋白质印迹法检测磷酸化NF-κB抑制蛋白激酶(phosphorylated inhibitor of NF-κB kinase,p-IKK)、磷酸化NF-κB p65(phosphorylated NF-κB p65,p-NF-κB p65)、NF-κB抑制蛋白α(inhibitor of NF-κBα,IκBα)蛋白表达水平变化,评价祛湿清肺方提取液抗炎活性。结果:星点设计-效应面法优化所得的最佳提取工艺为加水量13倍,提取2次,每次提取时间105 min。祛湿清肺方提取液能降低IL-6、IL-1β、TNF-α、NO的含量,抑制p-IKK、p-NF-κB p65蛋白表达,促进IκBα蛋白表达,具有较好的体外抗炎活性。结论:熵权法结合星点设计-效应面法优选的祛湿清肺方提取工艺稳定可行,所得提取液具有较好的抗炎活性,为祛湿清肺方开发和现代化研究提供参考奠定基础。     

胆淤方对胆汁淤积小鼠胆汁酸代谢的影响

目的 探讨胆淤方对1,4-二氢-2,4,6-三甲基-3,5-吡啶二甲酸二乙酯（DDC）诱导胆汁淤积小鼠血清胆汁酸代谢的影响。方法 40只雄性C57BL/6J小鼠随机分为4组：对照（N）组、DDC模型（M）组、奥贝胆酸（O）组、胆淤方（D）组,每组10只。N组小鼠用普通饲料喂养,M、O和D组小鼠用DDC饲料喂养,连续6周。造模第3周开始药物干预,O组ig予奥贝胆酸溶液（0.03 g·kg^-1）,D组ig予胆淤方药液（33 g·kg^-1）,M组和N组予相同体积的双蒸水。干预4周,收集小鼠血清和肝组织,试剂盒检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）、总胆红素（TBIL）和总胆汁酸（TBA）水平,HE染色观察肝组织炎症反应,CK19染色观察细胆管增生反应,天狼腥红染色观察纤维化,采用液质联用技术检测血清胆汁酸谱。结果 O组和D组ALT、AST、ALP、TBA、TBIL水平均较M组显著下降（P＜0.05）;D组ALT、AST水平较O组显著下降（P＜0.05）。D组小鼠肝组织炎症反应、纤维化及细胆管增生较M组减轻。D组血清初级胆汁酸胆酸（CA）和ω-鼠胆酸（ω-MCA）水平均较M组显著下降（P＜0.001）;D组血清次级胆汁酸甘氨胆酸（GCA）、牛磺胆酸（TCA）、牛磺熊去氧胆酸（TUDCA）、牛磺鹅去氧胆酸（TCDCA）、牛磺去氧胆酸（TDCA）、牛磺猪去氧胆酸（THDCA）较M组显著下降（P＜0.05、0.01、0.001）,O组的CA、ω-MCA、TCDCA、GCA和TCA水平较M组显著下降（P＜0.001）,D组血清TUDCA水平较O组下降更加显著（P＜0.05）。热图分析显示13种胆汁酸可将M与N组、O与M组、D与M组、D与O组血清胆汁酸谱两两区分。结论 胆淤方可改善肝内炎症反应,具有广泛的降胆汁酸作用。     

基于HPLC指纹图谱及定量测定的益气逐瘀利水方质量评价研究

目的：建立益气逐瘀利水方的HPLC指纹图谱,并测定4种成分含量,为益气逐瘀利水方产业化开发的质量控制提供参考。方法：采用Agilent ZORBAX Eclipse XDB-C₁₈色谱柱,乙腈-0.1%磷酸水为流动相,梯度洗脱,体积流量1.0 mL·min^-1,柱温30 ℃,检测波长245 nm,建立10批益气逐瘀利水方指纹图谱。采用中药色谱指纹图谱相似度评价软件（2012年版）进行相似度评价,结合聚类分析（CA）、主成分分析（PCA）及正交偏最小二乘法-判别分析（OPLS-DA）模式识别技术进行质量评价,同时进行含量测定。结果：10批益气逐瘀利水方标准汤剂指纹图谱相似度均大于0.995,标定出共有峰24个,指认其中的4个共有峰（14号峰毛蕊异黄酮葡萄糖苷、15号峰阿魏酸、17号峰汉防己乙素、19号峰粉防己碱）,CA、PCA及OPLS-DA将10批样品分成2类。定量分析方法学考察结果良好,10批样品中毛蕊异黄酮葡萄糖苷、阿魏酸、汉防己乙素、粉防己碱的定量结果分别为43.98~64.18、107.32~167.95、122.63~175.21、391.62~582.02 μg·g^-1。结论：所建立的益气逐瘀利水方指纹图谱及定量测定方法稳定性好、重复性高,可更加全面系统地评价益气逐瘀利水方的质量。     

清热利湿方防治兔胆色素结石形成的实验研究

目的动态观察清热利湿方对兔胆色素结石模型胆汁成分的影响。方法取健康家兔40只,随机分为对照组,细菌感染组,清热利湿中药大、小剂量组;造成胆色素结石模型,观察胆汁中总胆红素（TB）、游离胆红素（UCB）、总胆汁酸（TBA）、钙离子（Ca^2＋）、细菌性和内源性β-葡萄糖醛酸酶（B-Gase）活性的变化。结果清热利湿中药能明显降低胆汁中TB、UCB、Ca^2＋含量（P〈0．05）,增加胆汁中总胆汁酸含量（P〈0．05）,并能降低胆汁中细菌性和内源性β—Gase活性（P〈0．05）。结论清热利湿方具有很好的降低胆汁成石性的作用。     

补肾化瘀祛痰方治疗多囊卵巢综合征的临床研究

目的了解补肾化瘀祛痰方治疗多囊卵巢综合征的临床疗效及可能的作用机制。方法以补肾化瘀祛痰中药为主的水煎内服方及外敷方（必要时联合西药二甲双胍）治疗多囊卵巢综合征46例,连续治疗6个月为1个疗程;观察治疗前后临床症状和体征（肥胖、多毛、痤疮及黑棘皮症评分）、血清激素水平及胰岛素释放试验的变化情况。结果治疗后患者多毛、痤疮、黑棘皮症评分值及肥胖评分值与治疗前比较均明显下降（P〈0．01,P〈0．05）;血清睾酮（T）水平较治疗前显著降低（P〈0．05）;胰岛素试验中空腹胰岛素值与治疗前比较下降（P〈0．01）,糖负荷后30min、60min胰岛素值及胰岛素曲线下面积（IAUC）与治疗前比较亦有明显下降（P〈0．05）。结论以补肾化瘀祛痰方可以显著改善多囊卵巢综合征患者的症状和体征,改善患者的卵巢功能。     

Study on clinical effect and mechanism of Jianpi Qingre Huayu Recipe (健脾清热化瘀方) in treating patients with gastric ulcer

Objective： To study the effects of Jianpi Qingre Huayu Recipe （健脾清热化瘀方, JQH） in curing gastric ulcer and to preliminarily probe into its pathogenic mechanism, Methods： Fifty patients with gastric ulcer of Pi （脾）-insufficiency and stasis-heat syndrome type were assigned to the treated group （30 patients） and the control group （20 patients）. They were treated respectively with JQH and Ranitidine. At the same time, another group consisting of 20 healthy persons was set up for normal control. The clinical effect on gastroscopic figure and traditional Chinese medicine （TCM） syndrome were observed. Changes of T-cell subsets and interleukin-8 （IL-8） in serum as well as IL-8 in mucosa around the gastric ulcer were determined before and after treatment by flow cytometry and ELISA. Results： Comparison of the total effective rate on gastroscopic figure in the treated group and the control group （86.7% vs 80.0%） showed insignificant difference, but the cure rate and markedly effective rate in the former （50.0% and 20.0%） was higher than that in the latter （40.0% and 15.0%） respectively. Comparison of the total effective rate on TCM syndrome in the treated group and in the control group （96.7% vs 70.0%） showed insignificant difference, but the cure rate and markedly effective rate in the former （63.3% and 23.3%） was higher than that in the latter （50.0% and 20.0%） respectively. Serum levels of CD3^＋, CD4^＋, CD8^＋ got restored to normal range in the treated group after treatment but it was not so in the control group. IL-8 level in gastric mucosa was improved in both groups but the improvement in the treated group was better. Cenclusien： JQH could effectively treat gastric ulcer and partly reduce its recurrence through improving patients＇ immune function.     

健脾祛痰化瘀方对氧化型低密度脂蛋白诱导血管细胞信号分子钙离子和蛋白激酶C表达的影响

为了探讨健脾祛痰化瘀方在抗氧化型低密度脂蛋白致动脉粥样硬化中对信号转导分子钙离子和蛋白激酶C的影响，分别以健脾祛痰化瘀方-沥水调脂胶囊含药血清(浓度分别为5％、10％和20％)、氧化型低密度脂蛋白(100mg／L)处理人脐静脉内皮细胞和人脐动脉平滑肌细胞，以流式细胞仪检测两种细胞胞质游离钙离子浓度，以液体闪烁仪检测平滑肌细胞胞膜蛋白激酶C活性。结果发现，沥水调脂胶囊含药血清对氧化型低密度脂蛋白引起的内皮细胞、平滑肌细胞胞质游离钙离子浓度升高及平滑肌细胞胞膜蛋白激酶C活性升高有明显的抑制作用，其中20％含药血清作用最为明显。结果提示，健脾祛痰化瘀方可能通过调节钙离子、蛋白激酶C两种信号传递分子的变化起抗氧化作用，进而抑制内皮细胞凋亡及平滑肌细胞增殖，达到阻止动脉粥样硬化发生的作用。     

引经药独活根合二妙方可促进骨关节炎软骨损伤大鼠骨髓造血干细胞归巢

Objective:To investigate the effect of Ermiao Recipe(二妙方,EMR)with medicinal guide Angelicae Pubescentis Radix(APR)on the homing of bone marrow stem cells(BMSCs)to focal zone in osteoarthritis(OA)rats.Methods:Forty-eight Sprague-Dawley rats were randomly assigned to the sham-operated,model,EMR,and EMR plus APR groups(12 rats in each group).The OA rat model was induced by anterior cruciate ligament transection and medial meniscus resection.All rats were injected with recombinant human granulocyte colonystimulating factor[rhG-CSF,30μg/(kg-d)for continuous 7 days],and rats in the EMR and EMR plus APR groups were treated with EMR or EMR plus APR at 1.6 or 1.9 g/(kg·d)for 3 or 6 weeks,respectively.Cartilage histopathologic changes were observed by hematoxylin and eosin staining.Chondrocytes apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining,lnterleukin-1 B(IL-1β),tumor necrosis factorα(TNF-α),bone morphogenetic protein 2(BMP-2),and transforming growth factor beta-1(TGF-β_1)in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay assay.Matrix metalloproteinase(MMP)-13,tissue inhibitors of metalloproteinase(TIMP)-1,bromodeoxyuridine(BrdU),cluster of differentiation 34(CD34),and stromal cell derived factor 1(SDF-1)were measured by immunohistochemistry assay.Results:EMR and EMR plus APR significantly inhibited articular cartilage damage and synovium inflammation in OA rats at 3 or 6 weeks of treatment,the most obvious changes in these parameters were found in the EMR plus APR group.At 6 weeks,compared with EMR treatment,EMR plus APR remarkably inhibited chondrocytes apoptosis and the release of IL-1βand TNF-α,obviously decreased MMP-13 expression,and significantly increased expressions of proteoglycan,collagen,typeⅡcollagen and TIMP-1,serum levels of BMP-2 and TGF-β_1 as well as expressions of BrdU,CD34 and SDF-1 in cartilage articular(P0.01 or P0.05).Conclusion:The medicinal guide APR improved the therapeutic effects of EMR on OA rats by promoting directional homing of BMSCs to focal zone.     

宣肺止嗽方治疗感染后咳嗽临床疗效观察

目的：探讨宣肺止嗽方治疗感染后咳嗽的临床疗效。方法：感染后咳嗽患者120例随机分成两组,治疗组70例,采用宣肺止嗽方治疗,7d为1个疗程,疗程2周;对照组50例,口服惠菲宁,10mL／次,3次／d,7d为1个疗程,疗程2周。结果：两组患者治疗后临床疗效比较,治疗组总有效率94．29％,对照组总有效率80％,差异显著（P〈0．05）。结论：宣肺止嗽方治疗感染后咳嗽的疗效优于惠菲宁。     

通络方药对2型糖尿病大鼠心肌和主动脉iNOS基因表达的影响

目的观察通络方药（Tongluo Recipe,TLR）对2型糖尿病大鼠心肌和主动脉诱导型一氧化氮合酶（iNOS）基因表达的影响,探讨TLR在2型糖尿病（type 2 diabetes,T2DM）的干预作用和应用价值。方法30只雄性健康清洁级SD大鼠随机分为3组：正常对照组、T2DM组、T2DM＋TLR干预组,每组10只。T2DM＋TLR干预组大鼠TLR按0．4g·kg^-1·d^1剂量灌胃,1次,d,持续给药12W后处死取心肌、主动脉组织待测。利用实时定量逆转录多聚酶链反应（RT-PCR）方法测定各组大鼠心肌、主动脉诱导型一氧化氮合酶iNOS的基因表达水平。结果与正常对照组比较,2型DM组大鼠心肌和主动脉iNOS的mRNA表达明显降低（P〈0．01、P〈0．05）,而T2DM＋TLR干预组大鼠心肌和主动脉iNOS的mRNA表达较T2DM组明显升高（P〈0．01）。结论TLR可以增加T2DM大鼠心肌、主动脉iNOS的mRNA表达,从而增加NO合成,可能对T2DM具有防治意义。