迭鞘石斛抗肿瘤作用研究

体外培养SMMC-7721和HL-7702细胞,以不同浓度的迭鞘石斛多糖、醇提物、水提物作为药物组,并设生理盐水组作为阴性对照,5-Fu组作为阳性对照,MTT法分别测定处理24h、48h、72h后的吸光度值,计算药物对细胞生长的抑制率.荷瘤小鼠分别腹腔注射迭鞘石斛3种提取物,连续注射10d,d11眼眶静脉取血,测定血清中MDA含量及SOD活力变化.体外实验发现浓度为160μg/mL的迭鞘石斛多糖作用48h,对SMMC-7721细胞的生长抑制作用最强,且对HL-7702细胞的毒害作用较小.体内实验发现,迭鞘石斛多糖能显著增强荷瘤小鼠血清中SOD活力(P<0.01),降低MDA含量(P<0.01).研究表明,迭鞘石斛多糖是一种具有潜力的抗肿瘤天然药物.图1表2参8     

不同LED光质对迭鞘石斛生理及品质的影响

设置6种光质,自然光(对照,N5)、100%红光+0%蓝光(R5)、100%蓝光+0%红光(B5)、80%红光+20%蓝光[RB(4:1)]、60%红光+40%蓝光[RB(3:2)]、40%红光+60%蓝光[RB(2:3)],测定不同处理时间迭鞘石斛的生理指标以及多糖、黄酮的含量.结果显示光质对迭鞘石斛生理及品质有较为明显的影响.B5处理组的超氧化物歧化酶含量最高;R5处理组的丙二醛含量最高.RB(3:2)处理组的叶绿素含量最高,RB(2:3)处理组的可溶性糖、可溶性蛋白、多糖含量都最高;N5处理组的黄酮含量最高.R5处理组的叶绿素、可溶性糖、可溶性蛋白、多糖和黄酮含量都较低,说明纯红光并不利于迭鞘石斛生长.此外,不同处理天数下丙二醛、可溶性蛋白、可溶性糖、多糖和黄酮含量整体上呈现上升趋势;超氧化物歧化酶活力整体下降趋势;叶绿素含量整体上呈现先下降后上升的趋势.本研究说明红蓝光配比有利于迭鞘石斛生长发育和次生代谢物质积累,对迭鞘石斛组培苗的光控培养和品质改良具有生产上的指导意义.     

迭鞘石斛中性多糖DDP1-1的体内免疫活性

建立S180肉瘤模型,考察了迭鞘石斛中性多糖的体内免疫活性.以不同浓度的多糖作为药物组,环磷酰胺为阳性对照,生理盐水为阴性对照.给药14d后处死并解剖小鼠,眼眶取血,计算抑瘤率和免疫指数,检测血清中TNF-α、IL-2和IFN-γ含量.结果显示不同浓度的多糖表现出不同的免疫活性,其中多糖低浓度组(1.25mg/mL)有较高的肿瘤抑制率和免疫指数(P0.01),并促进和调节IL-2、IFN-γ、TNF-α在机体内分泌(P0.01),协调改善机体免疫系统功能,达到治疗肿瘤的目的.     

石斛不同极性成分的提取及其对肺癌细胞的抑制作用

为探究石斛(Dendrobium)的不同极性成分对人肺癌细胞抑制作用的差异,寻找到石斛抗癌活性成分高效提取的方法,采用极性不同的溶剂和方法对金钗石斛(D.nobile)、迭鞘石斛(D.denneanum)的药用成分分别进行提取以及层析柱纯化,以CCK-8实验检测不同极性提取物对人肺癌细胞和正常细胞的抑制作用,同时采用傅里叶变换红外光谱仪分别对石斛的乙醚和乙醇提取物成分进行检测和比较.结果显示,石斛的不同极性成分抑癌作用差异显著,石斛的抑癌成分极性较小,易溶于乙醚.经石油醚预洗后的石斛乙醚提取物对肺癌细胞的抑制作用较强,100μg/mL的金钗石斛与迭鞘石斛提取物对肺癌A549细胞的24 h抑制率分别为45%和40%.而传统中药成分提取中常用的水和乙醇对石斛抗癌成分的提取效率较低.红外光谱分析亦显示石斛的乙醚提取物与乙醇提取物化学成分上存在较大差别,因此对于石斛抗癌成分的提取应采用极性较小的乙醚等溶剂.此外,发现金钗石斛的特定极性组分C具有较强的抑制肺癌细胞作用,其100μg/mL的浓度对肺癌A549细胞的24 h抑制率达到97%,可作为石斛抗癌新药筛选的来源.因此,金钗石斛和迭鞘石斛的抑癌成分具有特定的极性,结果可为石斛抗癌成分及抗癌作用的进一步研究和利用提供参考.     

迭鞘石斛的化学成分

迭鞘石斛(Dendrobium denneanum)为川产石斛主流品种,在传统中医药中应用广泛,主要具有促进肠胃运动、抗肿瘤、增强机体免疫能力、扩张血管、抗氧化等药理作用.现从迭鞘石斛(Dendrobium denneanum)茎中分离得到4个化合物,其结构经波谱方法确定为鸟苷(1)、它乔糖苷(2)、vanilloloside(3)和9-β-D-allofuranulsylguanine(4).这4个化合物均为首次从迭鞘石斛中分离得到,其中它乔糖苷(2)和9-β-D-allofuranulsylguanine(4)为首次从该属分离得到.     

基于定量蛋白质组学分析迭鞘石斛对肺癌细胞的抑制作用及机理

以基于SILAC(Stable isotope labeling with amino acids)的定量蛋白质组学为主要研究方法,通过精确定量蛋白质含量并进行差异分析,从蛋白层面系统研究迭鞘石斛(Dendrobium denneanum)乙醇提取物对人肺癌细胞A549的影响.结果发现迭鞘石斛乙醇提取物对A549细胞具有明显的抑制作用,其在降低细胞增殖速率的同时导致细胞发生死亡,24 h和48 h时的抑制率分别为35.05%和59.08%.定量蛋白质组学实验发现迭鞘石斛乙醇提取物引起A549细胞中392个蛋白质的含量出现明显下调.功能分析显示下调蛋白主要包括细胞物质和能量代谢相关蛋白,如RNA代谢与蛋白质合成、糖类代谢、脂类代谢、嘧啶与嘌呤代谢等.同时大量与细胞抵抗内源及外界不利刺激和损伤相关的蛋白,如具有氧化还原、分子伴侣、DNA修复、蛋白降解与修复等功能的蛋白含量也出现明显的下调.本研究通过定量蛋白质组学实验从全细胞蛋白层面系统解释了迭鞘石斛的抑癌作用机理,可促进石斛抗癌机理的进一步研究和石斛源抗癌新药的开发工作.     

MDMA(3,4亚甲基二氧甲基苯丙胺)对小鼠睾丸细胞微核率及染色体畸变率的影响

通过连续20 d对雄性小鼠灌胃染毒3,4-亚甲基二氧基甲基苯丙胺(3,4-methylenedioxymethamphetamine,MDMA)后,探究MDMA对雄性小鼠睾丸组织细胞微核率及染色体畸变率的影响.将雄性小鼠随机分为MDMA低(5.0mg/kg)、中(10.0mg/kg)、高(20.0 mg/kg)三个染毒剂量组,采用生理盐水做阴性对照,每日染毒1次.于末次给药后第二天,取小鼠睾丸组织细胞,采用常规微核(micronucleus,MN)试验,检测小鼠睾丸细胞微核率的改变;同时采用染色体畸变试验(chromosomal aberration test)探究MDMA对小鼠睾丸细胞染色体畸变率的影响.微核试验结果表明MDMA中、低剂量组小鼠睾丸细胞微核率与阴性对照组比较差异无显著性(P0.05),而高剂量与低剂量组小鼠睾丸细胞微核率及阴性对照组比较,差异均有显著性(P0.01).染色体畸变试验结果中,MDM高、中剂量组染色体畸变率分别与阴性对照组比较差异有显著性(P0.05),剂量组之间染色体畸变率差异也有显著性(P0.05).结论通过微核试验与染色体畸变试验结果同时得出:在该实验剂量内MDMA高剂量组能诱导小鼠睾丸组织细胞微核率增加,高剂量组及中剂量组可以使小鼠睾丸细胞染色体畸变率增高,说明其对雄性小鼠睾丸遗传物质有一定的损伤效应.图4,表2,参10.     

长程高脂肪膳食对大鼠空腹血糖及糖耐量的影响

研究了单纯高脂膳食喂养对大鼠空腹血糖(Fasting plasma glucose,FPG)及葡萄糖耐量(Glucose tolerancetest,GTT)的影响,旨在模拟与现代人生活饮食结构及发病情况更为接近的糖调节异常动物模型.取正常SD大鼠100只喂以普通饲料或高脂饲料14mo,动态监测动物体重及空腹血糖的变化,并分别在第5、8、10、14mo时用糖耐量试验观察葡萄糖耐量减退(IGT)的情况.结果显示,高脂喂养第1~14个月过程中,雄性大鼠的体重较普食组均未出现显著增加,而雌性大鼠体重仅在喂养第5~8个月期间较普食组增加了10%~15%;与普食组相比,在喂养高脂膳食第1~5个月中雌雄两性大鼠空腹血糖均显著升高,在喂养第5~10个月中雄性葡萄糖耐量显著减退,但雌性动物则未出现明显的IGT,至第14个月时雄性IGT与普食组的差异消失.研究表明,单纯高脂膳食虽不能诱发糖尿病,但可出现较典型的IFG空腹血糖受损和IGT现象,且以雄性动物造模更为适宜,喂养时间以10mo以内为宜.由于单纯高脂喂养更接近自然人群发病过程,因此,此种模型将为糖调节受损发生机制及其早期干预的研究提供新的动物模型.图2表1参12     

多拷贝表达转鲑鱼降钙素基因酵母的急性、亚急性毒性

口服转鲑鱼降钙素基因酵母为鲑鱼降钙素的应用开辟了一条新的途径.为评价转鲑鱼降钙素基因酵母的安全性,将转基因酵母分别灌胃昆明种小鼠和Wistar大鼠进行急性毒性实验(7 d)、亚急性毒性实验(8周).急性毒性实验结果表明灌喂转基因酵母对小鼠无致死效应(LD50>10 000 mg/kg);亚急性毒性实验中高(2.0 g/kg)、低(0.5 g/kg)剂量组动物的一般状况、体重、主要脏器系数、血液学指标及血液生化指标与各对照组比较,均未见明显差异(P>0.05),组织病理切片检查未发现病理性变化.实验结果说明转鲑鱼降钙素基因酵母不会对实验动物造成不良影响,是安全无毒的.图3表3参16     

白眉蝮蛇（Agkistrodon halys ussuriensis）蛇毒精氨酸酯酶对小鼠的生殖毒性及F1子代的影响

研究了白眉蝮蛇蛇毒中分离的纯精氨酸酯酶对小鼠的一般生殖毒性及对F1代小鼠的影响 .0 .36U/kg剂量使雄性F0 小鼠体重增长缓慢 ,并使睾丸系数增大 (P <0 .0 5 ) ,但病理学检验睾丸和附睾正常 ;该剂量处理孕鼠 ,减轻了胎鼠和新生F1代小鼠的体重 (P <0 .0 5 ) ,但对F0 雌性小鼠的生殖功能及F1代小鼠的一般体征、中枢神经系统发育及生殖机能均无影响 (P >0 .0 5 ) .纯精氨酸酯酶对小鼠生殖无毒剂量为 0 .18U/kg .表 2参 9     

Antihyperglycemic and antihyperlipidemic effects of Tephrosia purpurea leaf extract in streptozotocin induced diabetic rats

Diabetes mellitus is a worldwide leading metabolic syndrome, associated with profound alterations in carbohydrate, lipids, lipoproteins and protein metabolisms. Worldwide, traditional practitioners for the treatment of diabetes and its complications use a wide variety of medicinal plants. In the present study the aqueous extract of Tephrosia purpurea leaves (TpALet) was evaluated for its antihyperglycemic and antihyperlipidemic effects in streptozotocin induced diabetic rats. Profound alterations in the concentrations of blood glucose, lipids and lipoproteins were observed in diabetic rats. Oral administration of TpALet to diabetic rats at a dose of 600 mg/kg body weight significantly reduced the level of blood glucose and increased the level of plasma insulin as well as normalized the lipids and lipoproteins profile. The present study thus demonstrated that TpALet has prominent antihyperglycemic and antihyperlipidemic effects in streptozotocin induced diabetic rats.     

大气细颗粒物与糖尿病

随着我国社会经济和城市化的快速发展,大气颗粒物是影响我国城市空气质量的首要污染物,大气细颗粒物污染已严重威胁我国居民健康。本文简要综述了PM2.5诱导的氧化应激和炎症反应在糖尿病的发生和发展中的分子作用机制。     

Phytoconstituents evaluation and antihyperglycemic and antihyperlipidemic effects of Mahonia leschenaultia Takeda in streptozotocin-induced diabetic rats

Hyperlipidemia is an associated complication of diabetes mellitus. Many spices and herbs are known to be hypoglycemic. Mahonia leschenaultia Takeda (Berberidaceae) (MLT) is a traditional medicinal plant that is commonly used against diseases in India and other Asian countries. The aim of this study is to identify the phytoconstituents by gas chromatography–mass spectrography (GC–MS) and evaluate the antihyperglycemic and antihyperlipidemic effects of the ethanol extract of MLT on streptozotocin (STZ)-induced diabetic rats. Phytoconstituents like oleicacid, 9,12-octadecadienoic acid (Z, Z)-n-hexadecanoic acid were identified as high % peak when compared with other compounds by GC–MS. STZ-induced diabetic animals were fed with plant extracts at different doses (250 or 500 mg kg^?1 body wt) of ethanol extract of MLT. The results of the study revealed a significant increase in total hemoglobin and glycosylated hemoglobin along with the reduction in hyperglycemia and hyperlipidemia effects in MLT-treated STZ-induced diabetic rats compared to diabetic only rats. Further, ethanol extracts of MLT-treated animals at 500 mg kg^?1 showed that shrinkage of β cells of islets of Langerhans was restored as evidenced by histological studies of pancreas of diabetic animals. Thus the ethanol extract of MLT might serve as a reliable adjuvant for antihyperglycemic and antihyperlipidemic effects of STZ, and may be promising for development of phytomedicines for diabetes mellitus.     

Interspecific and intraspecific identification of Dendrobium based on the psbAtrnH intergenic region sequences and the 5S rRNA gene spacer sequences北大核心CSCD

This research aimed to investigate the interspecific and intraspecific identification of Dendrobium by using the multi-locus method so as to provide a molecular basis for Dendrobium identification through the combination of chloroplast psbA-trnH intergenic region sequences and ribosome 5S rRNA gene spacer sequences. PCR direct sequencing was applied to detect the chloroplast psbA-trnH intergenic region sequences as well as the ribosome 5S rRNA gene spacer sequences of 12 Dendrobium species, while the psbA-trnH intergenic region sequences of Dendrobium denneanum dq-2 variety and dq- 5line were cloned and sequenced for single nucleotide polymorphism (SNP) analyzing. The sequences were analyzed by the software Sequencher4.14, Bioedit7.0, MEGA5.2 and Dansp5.0; the interspecific and intraspecific Kimara-2-Parameter(K2P) distances were also calculated. The phylogenetic tree (using Neighbor joining method) was constructed with Bulbophyllum odoratissimum and Bletilla striata as outgroup. The results showed an average length of chloroplast psbA-trnH gene sequences in Dendrobium as 742.3 bp, with 72 variable sites, including 33 information sites; the average length of the ribosome 5S rRNA gene spacer sequences in Dendrobium was 336.4 bp, with 213 variable sites including 139 information sites. Using psbAtrnH intergenic region sequences in combination with ribosome 5S rRNA gene spacer sequences can not only identify D. denneanum, D. hancockil, D. thysiflorum, D. devonianum, D. moniliforme, D. chrysotoxum, D. officinale, D. heterocarpum and D. nobile, but also differentiate D. officinale from different geographical populations, and distinguish the dq-2 variety and dq 5line with SNP in the multi locus of D. denneanum.     

光生物反应器培养紫球藻及藻粉降血脂的研究

研究了搅拌式光生物反应器中搅拌速度、通气量对紫球藻生长、胞外多糖和藻细胞超微结构的影响．实验结果表明：在特定光照(1000μmolm^-2s^-1)和培养温度(25℃)条件下，当搅拌转速为200r／min、通气量为0．71vvm时，藻生长速度和胞外多糖含量最高；增加通气量有利于藻细胞胞外多糖及生物量的提高．对藻细胞超微结构的观察发现，与摇床培养相比，搅拌式光生物反应器使藻细胞淀粉颗粒明显增多，粘质鞘层变薄，光合片层增多．动物实验表明，紫球藻能明显降低摄食高脂饲料SD大鼠血脂TC、TG、LDL-C水平，并提高HDL-C含量．实验结果预示，紫球藻对预防动脉粥样硬化和冠心病的发生有一定的意义．图2表3参26     

The cholesterol lowering property of coriander seeds (Coriandrum sativum): mechanism of action

Coriandrum sativum (Coriander) has been documented as a traditional treatment for cholesterol and diabetes patients. In the present study, coriander seeds incorporated into diet and the effect of the administration of coriander seeds on the metabolism of lipids was studied in rats, fed with high fat diet and added cholesterol. The seeds had a significant hypolipidemic action. In the experimental group of rats (tissue) the level of total cholesterol and triglycerides increased significantly There was significant increase in beta-hydroxy, beta-methyl glutaryl CoA reductase and plasma lecithin cholesterol acyl transferase activity were noted in the experimental group. The level of low density lipoprotein (LDL) + very low density lipoprotein (VLDL) cholesterol decreased while that of high density lipoprotein (HDL) cholesterol increased in the experimental group compared to the control group. The increased activity of plasma LCAT enhanced degradation of cholesterol to fecal bile acids and neutral sterols appeared to account for its hypocholesterolemic effect.     

Effect of Aegle marmelos and Hibiscus rosa sinensis leaf extract on glucose tolerance in glucose induced hyperglycemic rats (Charles foster)

In an effort to test the hypoglycemic activity of Aegle marmelos and Hibiscus rosa sinensis in glucose induced hyperglycemic rats, their alcoholic leaf extracts were studied. Both the groups of animals receiving either. A. marmelos or H. rosa sinensis leaf extract for seven consecutive days, at an oral dose equivalent to 250 mg kg-1 showed significant improvements in their ability to utilize the external glucose load. Average blood glucose lowering caused by A. marmelos and H. rosa sinensis was 67% and 39% respectively, which shows that former significantly (p < 0.001) improves the glucose tolerance curve. The magnitude of this effect showed time related variation with both the plants. Efficacy of A. marmelos and H. rosa sinensis was 71% and 41% of glybenclamide, respectively. These data throw some light on the possible mechanism of hypoglycemic activity of both the plants. The mechanism of action could be speculated partly to increased utilization of glucose, either by direct stimulation of glucose uptake or via the mediation of enhanced insulin secretion.     

Effect of hypoxia on the circulating levels of essential mineral elements in rats

Changes occurring in concentrations of certain trace metals and electrolytes viz. chromium, copper, zinc, sodium, potassium, calcium, magenesium and chloride in plasma of rats exposed to intermittent hypobaric hypoxia were evaluated. Batches of Sprague-Dowley rats (12 in each group) were exposed for 1, 7,14 and 21 days to a simulated altitude 7,620 m for 6 h per day and one group of unexposed animals was kept as control. There was a significant rise of 153% in plasma chromium levels of 1 day exposed group in comparison to the unexposed group which tends to normalise on subsequent exposure. There was a gradual increase in plasma copper levels of 9.0, 28.2, 62.6 and 65.6% respectively in 1,7,14 and 21 days exposed rats in comparison to unexposed rats. On the other hand plasma zinc levels were seen to be decreasing during entire exposure. Plasma sodium levels decreased initially in 1 and 7 day exposed rats and increased in later groups whereas plasma potassium levels of exposed groups remained low in comparison to unexposed group. Chloride levels were found to be elevated in 14 and 21 day exposed groups. The plasma calcium and magnesium levels were higher in all exposed groups over unexposed groups. Changes in chromium, copper and zinc observed in the present study during exposure to hypoxic stress may be responsible for the hyperglycemia and anorexia encountered during intial phase of high altitude acclimatisation.     

Relationships between exposure to polyhalogenated aromatic hydrocarbons and organochlorine pesticides and the risk for developing type 2 diabetes: a systematic review and a meta-analysis of exposures to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

A systematic review was performed to evaluate the association between environmental exposures to polyhalogenated aromatic hydrocarbons (PHAHs) and organochlorine (OC) pesticides and the risk for type 2 diabetes (T2D). Searches of EMbase, Google Scholar, Medline, and Scopus were performed. Reports were included if they were original human studies whose design included a control group and if the sample size was larger than 10 participants per group. The articles were excluded if the type of diabetes was unknown or Type 1. Odds ratios (OR), 95% confidence intervals (CI), and modified Naranjo scores for the effects of environmental contaminants were determined for each study. Of the 35 eligible studies, six were included in the meta-analysis; these focused specifically on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Meta-analysis of these studies showed no significant increased OR for the development of T2D from exposure to TCDD. However, subgroup analysis showed significant elevated OR for the development of T2D if exposure is repeated and accompanied by exposure to other persistent pesticides (OR?=?1.48; 1.10–1.90) but a decreased odds for T2D with exposure resulting from accidental exposure (OR?=?0.46; 0.39–0.52). Our results suggest that there are significant risks of developing T2D in populations with recurring exposures to TCDD in concert with other persistent chlorinated pesticides. A mean Naranjo score of 2.0 was determined for all 35 articles; this score was 2.5 when examining only TCDD studies and 2.1 when examining studies that looked only at T2D. Each of these Naranjo scores suggests a possible association between the prevalence of T2D and exposure to PHAHs or OC pesticides.