Effect of retention enema by caulis sargentodoxae enema combined with oral drugs on the immune inflammatory response and intestinal flora of UC patients

Objective:To study the effect of retention enema by caulis sargentodoxae enema combined with oral drugs on the immune inflammatory response and intestinal flora of UC patients. Methods: A total of 128 UC patients who received treatment in the hospital between January 2013 and April 2017 were collected and divided into control group and observation group by random number table. Control group received conventional oral drug therapy, and the observation group received retention enema by caulis sargentodoxae enema combined with oral drug therapy. Before treatment and after 4 weeks of treatment, the differences in the levels of immunoglobulin and inflammatory response indexes in serum as well as intestinal flora levels in mucosa lesion tissue were compared between the two groups.Results: The differences in levels of immunoglobulin and inflammatory response indexes in serum as well as intestinal flora levels in mucosa lesion tissue were not statistically significant between the two groups of patients before treatment. After 4 weeks of treatment, IgA, IgG, IgM and IL-4 contents in serum as well as bifidobacterium and lactobacillus levels in mucosa lesion tissue of observation group were higher than those of control group while ESR, CRP, IL-6 and TNF-αcontents as well as E. coli level in mucosa lesion tissue were lower than those of control group. Conclusion: Retention enema by caulis sargentodoxae enema combined with oral drugs can effectively optimize the immune inflammatory state and balance the intestinal flora distribution in patients with UC.

A new macrolide and glycosides from the stem of Sargentodoxa cuneata

新 macrolide glycoside， cuneataside F 从 Sargentodoxa cuneata 的茎的 n-butanol 摘录被孤立。结构象 HRESIMS 分光镜的分析一样根据广泛的 1D 和 2D NMR 被阐明。

Effects of “Caulis Sargentodoxae Formula” on Experimental Endometriosis in Rat

Objective To study the drug effects of “Caulis Sargentodoxae Formula” which clearsaway heat and blood clot on experimental endometriosis in rat, and to compare thiseffect with Danazol’s effects.Method The model of endometriosis rat was induced by transplanting with endometriumsurgically. The rats were divided into 6 groups randomly: the non-treatment group, thecastrate group, Danazol group(80 mg/kg), “Caulis Sargentodoxae Formula” group(110 g/kg), the medium dose group(77.5 g/kg) and the low dose group(55 g/kg). After3 weeks’ treatment, the volume of endometrium was detected by cutting the rats’ bellyopen, drawing the materials from endometrium for tissue section, and doing the quantityanalysis of endometrium by semiautomatic image analysis machine.Results There was no significant difference of the endometrium volumes between thelow dose group and the non-treatment group(P>0.05), other groups’ endometriumvolumes were significantly lower than those of the non-treatment group(P<0.05). Andapart from the low dose group, all other groups’ heights of the endometrium epitheliawere significantly lower than that of the non-treatment group(p<0.01).Conclusion “Caulis Sargentodoxae Formula” can refrain the growth of theendometrium, and the effects are better than that of Danazol’s.

Caulis Sargentodoxae Prescription Inhibits Angiogenesis-related Cytokines in a Rat Endometriosis Model

Background:To determine the efficacy of the Caulis Sargentodoxae prescription,which is an empirical formula of Chinese herbs and has definite curative effects on endometriosis.Methods:The Caulis Sargentodoxae prescription on the growth of ectopic endometria was studied with a rat endometriosis(EMS)model.The EMS model was established by autoplastic transplantation.To study the curative effects of Chinese medicine on EMS in comparison with western medicine,gestrinone and an angiogenesis inhibitor were introduced.The rats were randomly divided into seven groups:normal group,model group,ovariectomized group,gestrinone(western medicine)group,Caulis Sargentodoxae prescription(Chinese medicine)group,apatinib(inhibitor)group,and combination(Chinese medicine+inhibitor)group.After administration for 21 days,the growth inhibitory rates of ectopic endometria in the treatment groups were evaluated,and the levels of vascular endothelial growth factor(VEGF)and VEGF receptor-2(VEGFR2)were detected by ELISA in the serum and peritoneal fluid,as well as in the ectopic endometrial tissues by real-time polymerase chain reaction and Western blotting.Results:The growth inhibitory rates of ectopic endometria in the treatment groups were significantly higher(P<0.05).In the Caulis Sargentodoxae prescription group,the levels of angiogenesis-related factors,including VEGF and VEGFR2,were reduced in the serum and peritoneal fluid compared with the model group(P<0.05).In addition,the positive expression of VEGF and VEGFR2 in ectopic endometria significantly decreased in the Caulis Sargentodoxae prescription group both at mRNA and protein levels.Conclusions:VEGF and VEGFR2 levels in the serum and peritoneal fluid can be used as a clinical reference for endometriotic pathogenesis and treatment,and the Caulis Sargentodoxae prescription has reliable therapeutic effects on EMS for its target-action ability to decrease angiogenesis.

大血藤质子核磁共振指纹图谱研究

目的建立不同产地大血藤^(1)H-NMR指纹图谱,并进行差异性分析。方法采用CPMG脉冲序列采集样品^(1)H-NMR信号,测定温度298 K,观测频率600 MHz,数据经分段积分,归一化处理后导入MetaboAnalyst 5.0软件进行统计分析。结果建立了大血藤^(1)H-NMR指纹图谱,并鉴定了13种化学成分,主成分分析、聚类分析结果表明13批大血藤药材有显著差异,方差分析结果筛选出绿原酸乙酯、正十六烷酸、红景天苷、葡萄糖、3,5-O-二甲基没食子酸、丁香醛、蔗糖、表儿茶素、绿原酸为影响大血藤质量的主要差异性成分。结论建立了大血藤^(1)H-NMR指纹图谱并筛选出差异性成分,为评价不同来源大血藤药材的质量提供了参考。

红藤汤外敷联合中药保留灌肠治疗慢性盆腔炎临床研究

目的:观察红藤汤外敷联合中药保留灌肠治疗慢性盆腔炎的临床疗效。方法:将60例慢性盆腔炎患者按照随机数字表法分为对照组和观察组,每组各30例。两组均给予常规西药治疗,对照组予以红藤汤保留灌肠,观察组在对照组的基础上联合红藤汤外敷。比较两组患者的临床疗效及治疗前后中医证候积分、妇科检查情况、炎症因子[肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、C反应蛋白(C reactive protein,CRP)、单核细胞趋化蛋白-1(monocyte chemotactic protein-1,MCP-1)、粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)]、子宫血流动力学指标[子宫搏动指数(pulsation index,PI)、阻力指数(resistance index,RI)、最大峰值流速(maximum peak flow rate,PFE)]、血液流变学指标[血浆黏度(plasma viscosity,PV)、红细胞沉降率(erythrocyte sedimentation rate,ESR)、红细胞比容(hematocrit,Hct)]变化情况。结果:两组患者治疗后中医证候积分低于本组治疗前,且治疗后观察组低于对照组,差异具有统计学意义(P<0.05)。对照组有效率为70.00%,观察组有效率为93.33%,两组患者有效率比较,差异具有统计学意义(P<0.05)。两组患者治疗后宫颈触痛、子宫活动受限、宫体/附件压痛评分、炎性包块直径、盆腔积液最深径低于本组治疗前,且治疗后观察组低于对照组,差异具有统计学意义(P<0.05)。两组患者治疗后TNF-α、CRP、GM-CSF及MCP-1低于本组治疗前,且治疗后观察组低于对照组,差异具有统计学意义(P<0.05)。两组患者治疗后PV、ESR及Hct低于本组治疗前,且治疗后观察组低于对照组,差异具有统计学意义(P<0.05)。结论:红藤汤外敷联合中药保留灌肠治疗慢性盆腔炎,可改善患者的临床症状,抑制炎性反应,促进血液循环。

红藤水溶性部位的LC-MS分析及抗炎活性成分的虚拟筛选

目的:基于分子对接,确定中药红藤75%乙醇提取物的水溶性部位(HT_(W))中的抗炎物质基础及相关的炎症因子。方法:采用液质联用(LC-MS)技术确定HT_(W)中的主要化学成分。采用软件Autodock Vina对化合物与肿瘤坏死因子-α、白介素-1β转化酶、IL-6、IκB激酶β、诱导性一氧化氮合酶(iNOS)及Toll样受体2(TLR2)等6种靶点蛋白的结合能进行打分,虚拟筛选出抗炎活性成分以及关键靶点。结果:通过LC-MS分析,共确定14个含量较为丰富的小分子化合物,主要为苯丙酸、苯乙醇和木脂素类化合物,其中绿原酸类化合物含量最高。分子对接结果显示,TLR2与HT_(W)中化合物的结合能平均值最高,其中与化合物2、3、7、11-13的结合能均超过-9.0 kcal/mol,远超过阈值-7.2 kcal/mol。TLR2与木通苯乙醇苷B(12)的结合能为-9.7 kcal/mol,与绿原酸(3)及其异构体(2)亦有较好的结合力。iNOS对HT_(W)中化合物也较敏感,与苯乙醇苷类化合物10的结合能为-9.8 kcal/mol。结论:绿原酸、苯乙醇苷等化合物可能是红藤水溶性部位(HT_(W))最重要的抗炎活性物质;其抗炎关键靶点可能是TLR2和iNOS。

五花血藤水煎液通过抗炎作用促进脊髓损伤后小鼠运动功能恢复

目的探讨五花血藤水煎液(Sargentodoxa cuneata,SCD)在小鼠脊髓损伤(spinal cord injury,SCI)后的抗炎作用及其对运动功能恢复的影响。方法选取36只SPF级小鼠随机分成假手术组、脊髓损伤组和SCD组,利用BMS评分检测小鼠后肢运动功能恢复情况,免疫蛋白质印迹法检测各组小鼠脊髓白细胞介素(IL)-6、IL-12A、肿瘤坏死因子(TNF)-α和脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的表达,免疫荧光染色通过NeuN观察损伤处脊髓前角神经元变化,rAAV-retro逆行示踪观察各组小鼠胫骨前肌(tibialis anterior muscle,TA)相关脊髓运动神经元分布。结果与脊髓损伤组相比,SCD组小鼠在损伤后7 d的BMS评分显著提高(P<0.05),损伤后14 d,IL-6、IL-12A和TNF-α表达明显降低(P<0.05),BDNF的表达显著升高(P<0.05);损伤处脊髓前角神经元数量显著增多(P<0.05),TA相关运动神经元数量明显增多(P<0.05)。结论五花血藤水煎液能明显下调炎症因子IL-6、IL-12A和TNF-α的表达,发挥抗炎作用,同时上调BDNF的表达从而对运动神经元起保护作用,促进损伤后小鼠运动功能的恢复。

红藤败酱散加减联合氟罗沙星片治疗慢性盆腔炎临床研究

目的:观察红藤败酱散加减氟罗沙星片治疗慢性盆腔炎的临床疗效及其对患者炎性因子水平、血液流变学指标的影响。方法:将82例慢性盆腔炎患者按照随机数字表法分为观察组和对照组,每组各41例。对照组给予氟罗沙星片治疗,观察组在对照组治疗的基础上联合红藤败酱散加减。比较两组患者的临床疗效、不良反应发生率及治疗前后中医证候积分、炎性因子水平[白细胞介素(interleukin, IL)-1β、超敏C反应蛋白(hs-C reactive protein, hs-CRP)、IL-2、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)]、血液流变学指标[纤维蛋白原(fibrinogen, FIB)、血沉、全血低切黏度]变化情况。结果:两组患者治疗后中医证候积分低于本组治疗前,且治疗后观察组低于对照组(P<0.05)。观察组有效率为97.56%,高于对照组的82.93%(P<0.05)。两组患者治疗后hs-CRP、IL-1β、IL-2、TNF-α、FIB、血沉及全血低切黏度均低于本组治疗前,且治疗后观察组低于对照组(P<0.05)。观察组不良反应发生率为14.63%,对照组不良反应发生率为9.76%,组间比较,差异无统计学意义(P>0.05)。结论:红藤败酱散加减联合氟罗沙星片治疗慢性盆腔炎,可缓解患者的临床症状,降低炎性因子水平及血液流变学指标。

红藤饮片提取物抑菌活性与次生代谢产物含量的相关性

目的:了解3个不同产地、2个不同批次的红藤饮片6种次生代谢产物含量、抑菌活性及两者的相关性。方法:利用分光光度法对红藤饮片提取物的6种次生代谢产物含量进行分析,以金黄色葡萄球菌和枯草芽孢杆菌为实验菌株,用纸片琼脂扩散法测定抑菌圈大小;以平板二倍稀释法统计最小抑菌浓度(M IC);以试管二倍稀释法统计最小杀菌浓度(M BC)。结果:红藤药液对金黄色葡萄球菌和枯草芽孢杆菌的抑菌活性以安徽产地第一批次最高,抑菌圈大小、M IC、M BC分别为(9.67±0.29)mm、12.5 m g/m l、31.25 m g/m l和(10.17±0.58)mm、12.5 m g/m l、31.25 m g/m l,江苏产地第二批次最低,抑菌圈大小、M IC、M BC分别为(7.67±0.29)mm、25 m g/m l、125 m g/m l和(8.17±0.29)mm、25 m g/m l、125 m g/m l。红藤饮片提取物对金黄色葡萄球菌的抑菌活性在不同产地和不同批次之间均有显著性差异(P<0.01);对枯草芽孢杆菌的抑菌活性在不同产地亦有显著性差异(P<0.01),但不同批次之间无显著性差异(P>0.05)。红藤药液的6种次生代谢产物总量以安徽产地第一批次最高(3.61%),江苏产地第二批次最低(1.90%)。红藤提取物的总生物碱和游离蒽醌在不同产地之间有显著性差异(P<0.01),而在不同批次之间无显著性差异(P>0.05);总绿原酸、黄酮类化合物、总鞣质和总皂苷在不同产地和不同批次之间均有显著性差异(P<0.01)。影响红藤饮片对2种细菌的抑菌活性的主要次生代谢产物可能为总皂甙、总鞣质、游离蒽醌及总绿原酸。结论:不同红藤饮片的次代谢产物的含量及抑菌活性以安徽和浙江产地较高,江苏产地较低;红藤的抑菌活性与饮片中总皂甙、总鞣质、游离蒽醌及总绿原酸的含量密切相关。

大血藤黄酮类化合物组成的初步研究

大血藤为一种传统药用植物。为了了解大血藤的药用成分,利用聚酰胺薄层层析及HPLC法分析了大血藤各器官黄酮类化合物的组成。聚酰胺薄层层析及HPLC法分析都显示大血藤叶片含有10种不同的黄酮甙,而嫩茎、叶柄与老茎中分别含有10种、8种及5种不同的黄酮甙;大血藤叶片黄酮类化合物经盐酸水解后HPLC分析显示有8种不同的黄酮类化合物。大血藤叶片及嫩茎的黄酮类化合物组成成分较多,具有一定的应用开发前景。

中药大血藤中酚类化学成分及其抗肿瘤活性

目的阐明中药大血藤中抗癌活性成分。方法以细胞周期抑制、坏死性细胞毒活性为抗癌指标 ,采用硅胶、SephadexLH 2 0、大孔树脂HP 2 0等色谱方法 ,跟踪分离活性成分 ,利用理化性质及波谱方法鉴定了化学结构 ,用SRB法及流式细胞术评价其抗癌活性。结果与结论从大血藤茎中分离得到 7个酚酸类化合物 :3,5 O 二甲基 没食子酸 (1)、原儿茶酸 (2 )、绿原酸 (3)、N (对 羟基苯乙基 )阿魏酸酰胺 (4)、对 羟基苯乙醇 (5 )、(- ) 表儿茶素 (6 )、缩合鞣质B2 (7)。化合物 3对人慢性髓性白血病K5 6 2细胞的半数抑制浓度 (IC50 )为 97 2 μg/mL ,化合物 4在 10 0 μg/mL浓度下对K5 6 2细胞的增殖抑制率为 4 6 6 %。流式细胞术检测表明 ,化合物 7对小鼠乳腺癌tsFT2 10细胞和K5 6 2细胞均显示G2 /M期抑制作用 ,为一新的细胞周期抑制剂。除化合物 2外 ,其余 6个化合物均为首次从该属植物中分离得到。

不同海拔高度大血藤群体遗传多样性的RAPD分析及其与环境因子的相关性

利用 RAPD技术分析了分布于浙江省天台山 3个不同海拔高度的天然大血藤群体的遗传多样性、遗传分化以及与环境因子的相关性。 13种随机引物在 3 6株个体中共检测到 88个可重复的位点 ,其中多态位点 74个 ,总多态位点百分率为84.0 9% ,大血藤具有丰富的遗传多样性。 Shannon信息指数显示的遗传多样性以海拔 950 m的群体为最高 ,其次是海拔 73 0 m的群体 ,最低的是海拔 52 0 m的群体 ;群体内的遗传多样性占总遗传多样性的 43 .68% ,群体间的遗传多样性占 56.3 2 %。 Nei指数估计大血藤群体间的遗传分化系数为 0 .540 6,大血藤群体间的基因流很低。大血藤海拔 73 0 m群体与海拔 52 0 m群体的遗传相似度较高 ,海拔 950 m群体与其它两群体的遗传相似度较低。大血藤群体内的遗传多样性与土壤总氮呈极显著的正相关。

大血藤DNA提取及RAPD研究初探

以浙江天台山大血藤为材料 ,对其DNA提取和RAPD分子标记方法进行了研究。结果表明 :所采用的经改进的SDS提取法可获得高质量的大血藤DNA ,分子量大于 2 3kb ,可满足RPAD扩增 ;用 1 5个不同的随机引物对所提取的 1 2个个体的大血藤DNA进行了RAPD分子标记分析 ,其中 1 4个引物扩增产物具有多态性。建立了大血藤DNA提取和RAPD标记的分析程序 。

中国特有植物大血藤的RAPD分析──植物地带性分化的分子差异标准初探

采用随机扩增多态性ＤＮＡ技术对中国特有科特有属植物大血藤（Ｓａｒｇｅｎｔｏｄｏｘａｃｕｎｅａｔａ）进行分析，探讨植物地带性分化的分子差异．从１００个随机引物中筛选出１１个有效的１０碱基对寡聚核糖核苷酸作引物，用于对取自５省区的大血藤植物基因组ＤＮＡ进行ＰＣＲ扩增；用ＵＰＧＭＡ法对各居群间的Ｊａｃａｒｄ相似性系数和遗传距离进行聚类分析．ＲＡＰＤ分析结果表明：分布于各省区（地区）的大血藤居群，尽管其形态特征的变异尺度较为一致，但在分子水平上存在明显差异，这种差异与地理范围的变化相关，表现出一定的地带性分化规律．

中药红藤方对大鼠的异位子宫内膜芳香化酶表达的影响

目的:探讨中药红藤方对异位子宫内膜的抑制作用。方法:用手术自体移植法成功建立的子宫内膜异位症大鼠模型,随机分成5组:红藤方高剂量组(80g生药/kg,A组)、低剂量组(20g生药/kg,B组)、达那唑组(60mg/kg,C组)、去势组(D组)和模型组(E组),连续给药21d后,检测各组的血清E2水平及异位内膜和卵巢的芳香化酶的变化;另测定异位内膜的体积。结果:A组、C组和D组的异位内膜生长均明显受到抑制,且3组间无统计学差异(P>0.05),与E组比有统计学差异(P<0.05);组织学观察可见,异位内膜呈退化趋势,A、C和D组间大鼠血清中的E2水平无统计学差异(P>0.05),与E组相比有统计学差异(P<0.05);A组、C组及D组异位内膜、卵巢组织的芳香化酶P450低表达,A组与E组间有统计学差异(P<0.05)。结论:红藤方可通过抑制异位内膜、卵巢的芳香化酶活性表达,降低局部雌激素含量,抑制异位内膜的生长,使异位内膜萎缩。

大血藤黄酮类化合物的提取与分析

Several methods were used to extract the flavonoids from the leaves of Sargentodoxa cuneata while concentration of flavonoids in the extraction was determined by different methods. The suitable quantitative methods used in the ecological analysis of flavonids was that using ultraviolet and 70% ethanol was the best reagent for extraction. Heating was not necessary for extraction. The content of flavonoids from different organs was leafblade, annual twig, leafstalk and perennial stem in turn.

大孔树脂法分离纯化大血藤中原花青素

采用大孔树脂法提取和纯化大血藤中原花青素,以吸附及解吸附能力为指标,比较D101、HPD100、X5、AB8、及ADS17五种大孔树脂对原花青素的吸附效率,通过单因素实验考察上样流速、上样浓度、洗脱流速、洗脱剂用量及洗脱剂体积分数对提取原花青素含量的影响,优选树脂的动态吸附及解吸附条件,并评价提取得到原花青素的纯度。结果表明,HPD100树脂对大血藤中原花青素的吸附和解吸附效果最好,上样流速2 BV/h,上样浓度6 mg/m L,洗脱流速1 BV/h,洗脱剂用量2 BV的纯化效果最好,100%乙醇的洗脱量最大,得到原花青素的纯度是粗提物的1.76倍。

大血藤不同器官黄酮类化合物含量的季节变化

The contents of flavonoid in the leaf and stem of Sargentodoxa cuneata (Oliv.) Rehd. et Wils. were determined. In a whole growth season, the dynamic chang of flavonoid in the leafblade of S. cuneata showed the two-peak pattern. The first peak appeared at June and the second at September. The seasonal dynamic change of flavonoids in leafstalk, annual twig and perennial stem were different and inconspicuous. The flavonoid cumulated maximally in the leafblade and the followed was seccessively annual twig, leafstalk, and perennial stem. The flavonoid content in leafblade was 31.79 mg/g(DW), while that in annual twig, leafstalk and perennial stem was 5.41 mg/g(DW), 5.30 mg/g(DW) and 3.95 mg/g(DW) respectively. There was markedly higher flavonoid content in leafblade. It is considered that the leaf of Sargentodoxa cuneata is significant in further exploitation.

大血藤叶的生长及光合生理生态特性研究

对大血藤叶的生长及光合生理生态特性进行了研究 ,结果表明 :大血藤叶的长度、面积和干重的增加有共同的趋势 ,即在叶的生长早期增长很快 ,以后逐渐降低 ,叶长度、面积约在 7月中旬达到最大值 ,叶干重 8月初达到最大值。随着叶的生长 ,叶绿素含量、硝酸还原酶活力逐渐增加 ,至 8月达到最大值 ,以后逐渐下降。春、夏期间的光补偿点低于秋季 ,而光饱和点则高于秋季。叶的光合日进程在春季呈“单峰”曲线 ;在夏、秋季则呈“双峰”型 ,有明显的“午休”现象。日均净光合速率在整个生长季节的变化是随着叶的生长 ,先是上升 ,至 8月初达到最大值 ,然后逐渐下降。夏季 ,大血藤日均净光合速率比常绿阔叶树种低 。