栀子苷对正常大鼠急性肝、肾毒性的时-毒关系分析

目的:考察栀子苷对正常大鼠急性肝、肾毒性的时-毒关系,为栀子临床安全应用提供科学依据。方法:Wistar大鼠110只,随机分为正常组,给药后不同时间组(0.5,1,2,4,8,12,24,48,72 h组),除正常组灌服生理盐水外,其余组按剂量1.2 g·kg-1灌服栀子苷。按组在灌胃后相应时间眼眶静脉取血,离心取血清,检测血清天门冬氨酸氨基转移酶(AST),碱性磷酸酶(ALP),丙氨酸氨基转移酶(ALT),总胆红素(TBIL),尿素氮(BUN),肌酐(Cr)活性,观察肝肾毒性损伤情况。结果:与正常组比较,在给药12 h后AST,ALP,ALT,TBIL,BUN,Cr明显升高(P0.05,P0.01),在给药24,48 h后AST,ALP,ALT,TBIL,BUN,Cr出现峰值,72 h后明显下降,240 h可见基本恢复正常。病理组织学检查出现不同程度的汇管区炎细胞浸润、肝细胞坏死、汇管区胆管轻度增生、纤维组织增生等病理变化。结论:栀子苷(1.2 g·kg-1)对正常大鼠存在急性肝、肾毒性且存在一定的时-毒关系。     

Antinociceptive Effects of Eugenol Evaluated in a Monoiodoacetate‐induced Osteoarthritis Rat Model

The aim of the present study was to evaluate whether eugenol, the main constituent of clove oil, has the capacity to provide analgesia in the monoiodoacetate‐induced rat model of osteoarthritis. Animals (n = 6/group) received either eugenol (20 or 40 mg/kg) or a vehicle by gavage. Daily administrations were initiated 2 days post osteoarthritis induction and continued for the duration of the study (4 weeks). Gait analysis was performed using the CatWalk method and secondary mechanical allodynia was assessed with von Frey filaments. Selected spinal cord peptides (substance P, calcitonin gene‐related peptide and dynorphin) were quantified by mass spectrometry. Significant changes were identified in dynamic gait parameters (swing speed, swing phase duration and duty cycle) of the affected limb following 40 mg/kg eugenol treatment compared with the vehicle (p < 0.05). Von Frey results revealed significant differences between the 40 mg/kg treatment and the vehicle group during the first and the third week of the study (p < 0.02). Spinal pain‐related peptide analysis revealed a decreased content of substance P and CGRP accompanied by an increase of dynorphin in animals treated with 40 mg/kg eugenol. These results suggest a therapeutic potential of eugenol to alleviate osteoarthritis‐related pain. Copyright © 2012 John Wiley & Sons, Ltd.     

栀子苷对正常和黄疸模型大鼠的亚急性肝、肾毒性

目的:开展栀子苷对正常及黄疸模型大鼠的亚急性肝、肾毒性对比研究,为栀子临床安全用药提供科学依据。方法:SD大鼠80只,随机分为8组,空白组、栀子苷低、中、高剂量(60,180,360 mg·kg~(-1))组,黄疸模型组和α-萘异硫氰酸酯(ANIT)+栀子苷低、中、高剂量(60,180,360 mg·kg~(-1))组。黄疸模型采用ANIT造模,24 h后灌胃(ig)栀子苷。分别于给药后第7,14天,检测血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),总胆红素(TBIL),尿素氮(BUN),血肌酐(SCr)活性,取大鼠肝、肾组织进行病理检测。结果:与空白组比较,第7,14天栀子苷中、高剂量组TBIL,SCr明显升高(P0.05,P0.01);黄疸模型组大鼠ALT,AST,TBIL均明显升高(P0.05,P0.01)。与黄疸模型组比,第7,14天ANIT+栀子苷低、中剂量组AST显著升高(P0.01),ANIT+栀子苷中剂量组TBIL,BUN明显升高(P0.05)。ANIT+栀子苷高剂量组ig 7 d后陆续全部死亡。病理切片显示,栀子苷给药后肝肾组织出现极轻度炎细胞浸润和肾小管嗜碱性变;模型组和ANIT+栀子苷各剂量组动物均出现胆管上皮细胞增生,纤维组织增生,并且有随剂量增大病变程度加重的趋势。结论:栀子苷剂量超过180 mg·kg~(-1)(约折合栀子生药量55 g·d~(-1))连续ig给药14 d可对正常及黄疸模型大鼠造成肝、肾损伤;且剂量超过60 mg·kg~(-1)时即会加重黄疸模型大鼠已有的肝损伤,随剂量增加病变程度加重。     

小柴胡汤提取物灌胃给药的长期毒性实验研究

目的：观察小柴胡汤提取物长期连续灌胃给药对大鼠所产生的毒性反应。方法：小柴胡汤提取物2256，1128,100mg／kg灌胃给药和正常对照组作180天长期毒性试验，观察对大鼠的生长发育，血液学常规，血液生化指标和主要脏器病理形态的影响。结果：对大鼠生长发育，血液学常规，血液生化指标均在正常范围，主要脏器病理形态未出现毒性病变。结论：小柴胡汤提取物长期灌胃未发现毒性反应，长期服用安全。     

An Investigation of the Antidepressant Action of Xiaoyaosan in Rats Using Ultra performance Liquid Chromatography–Mass Spectrometry Combined with Metabonomics

A rapid, highly sensitive, and selective method was applied in a non‐invasive way to investigate the antidepressant action of Xiaoyaosan (XYS) using ultra performance liquid chromatography–mass spectrometry (UPLC‐MS) and chemometrics. Many significantly altered metabolites were used to explain the mechanism. Venlafaxine HCl and fluoxetine HCl were used as chemical positive control drugs with a relatively clear mechanism of action to evaluate the efficiency and to predict the mechanism of action of XYS. Urine obtained from rats subjected to chronic unpredictable mild stress (CUMS) was analyzed by UPLC‐MS. Distinct changes in the pattern of metabolites in the rat urine after CUMS production and drug intervention were observed using partial least squares–discriminant analysis. The results of behavioral tests and multivariate analysis showed that CUMS was successfully reproduced, and a moderate‐dose XYS produced significant therapeutic effects in the rodent model, equivalent to those of the positive control drugs, venlafaxine HCl and fluoxetine HCl. Metabolites with significant changes induced by CUMS were identified, and 17 biomarker candidates for stress and drug intervention were identified. The therapeutic effect of XYS on depression may involve regulation of the dysfunctions of energy metabolism, amino acid metabolism, and gut microflora changes. Metabonomic methods are valuable tools for measuring efficacy and mechanisms of action in the study of traditional Chinese medicines. Copyright © 2012 John Wiley & Sons, Ltd.     

益气养阴活血方对2型糖尿病大鼠模型降糖降脂作用研究

目的观察益气养阴活血方对2型糖尿病大鼠模型降糖、降脂作用研究。方法将50只SPF级雄性SD大鼠随机取10只作为空白对照组,其余大鼠随机分为模型4周组、模型12周组、给药4周组、给药12周组,每组10只大鼠。模型对照组、空白组灌胃给予蒸馏水,益气养阴活血方组每天上午灌服用蒸馏水稀释的益气养阴活血方,给予18 g生药/kg,10 mL,分别于给药后4周,于干预治疗后第4周末,模型对照4周组及益气养阴活血方4周组进行标本采集,模型12周组和给药12周组给药12周后于第12周末用同样的方法进行血液标本采集。结果益气养阴活血中药对用药12周糖尿病大鼠具有降糖作用(P<0.001);给药4、12周能降低糖尿病大鼠三酰甘油含量水平(P<0.05),对胆固醇、高密度脂蛋白、低密度脂蛋白含量水平的影响不明显(P>0.05)。结论益气养阴活血方可通过降低血糖、三酰甘油来控制2型糖尿病及其并发症的发生发展,需要长期服用,短期服用治疗效果不明显。     

不同频率针刺内关穴对MCAo模型大鼠脑血流量的影响

目的:研究不同频率针刺内关穴对大脑中动脉阻塞模型(middle cerebral artery obstruction,MCAo)大鼠脑血流量的作用及机制。方法:参照Zea-longa线栓法复制MCAo模型。成年雄性Wistar大鼠144只随机分为正常组、非针刺组、非穴组、内关组,后两者又按60次/分钟、120次/分钟、180次/分钟各分3组。针刺每次行针5秒钟,每天2次,共3天。观察指标为脑血流量、软脑膜微循环管径和脑组织微血管数。结果:针刺内关穴比非穴能明显提高MCAo模型大鼠脑血流量,扩张软脑膜微循环管径,增加脑组织微血管数,差异有统计学意义(P<0.01),且内关120次/分钟组、180次/分钟组与60次/分钟组比较,差异有统计学意义(P<0.01)。结论:针刺内关穴能提高MCAo大鼠局部脑血流量且其提高脑血流量作用可能与扩张软脑膜微循管径,增加脑组织微血管数有关。与非穴相比,针刺内关对MCAo大鼠脑血流量的影响作用具有特异性且与不同针刺频率有关,在行针时间为5秒前提下,120次/分钟、180次/分钟比60次/分钟频率针刺更能提高其脑血流量。     

胶原诱导性关节炎大鼠滑膜基因表达谱特征研究

目的:运用基因芯片技术研究胶原诱导性关节炎(collagen-inducing arthritis,CIA)大鼠滑膜基因表达谱特征,探讨CIA相关致病基因。方法:选用健康雄性Wistar大鼠40只,随机分为造模组、正常对照组(n=8,简称正常组),造模组采用牛Ⅱ型胶原乳剂于大鼠尾根部注射,建立CIA模型。在造模成功大鼠中随机选取8只作为模型组(n=8)。于造模成功后30天,提取各大鼠滑膜总RNA,运用Illumina基因表达谱芯片分析每个样本基因变化。结果:正常组与模型组大鼠滑膜差异表达基因有271条,其中上调基因131条,下调基因140条。差异表达基因主要涉及细胞凋亡、基质金属蛋白酶、癌基因、热休克蛋白、骨桥蛋白、细胞周期、代谢、信号传导通路、趋化因子、黏附分子等。结论:CIA是一涉及多基因表达异常的自身免疫性疾病,筛选到的差异基因如上调表达基因Map3k11、Mmp8、OPN5、Rgs5、Hsph1、Ccl20、Robo4、Hyal4及下调表达基因Cabp1等,有利于阐释RA分子机制,提供RA防治新靶点。     

Acute and Sub‐Acute Oral Toxicity Assessment of the Hydroalcoholic Extract of Withania somnifera Roots in Wistar Rats

Withania somnifera is a widely used medicinal plant for several disorders. Toxicity studies on Withania somnifera are not available. Acute and sub‐acute oral toxicities of Withania somnifera root extract in Wistar rats were evaluated in the present study. In the acute toxicity study, WSR extract was administered to five rats at 2000 mg/kg, once orally and were observed for 14 days. No toxic signs/mortality were observed. In the sub‐acute study, WSR extract was administered once daily for 28 days to rats at 500, 1000 and 2000 mg/kg, orally. No toxic signs/mortality were observed. There were no significant changes (P < 0.05) in the body weights, organ weights and haemato‐biochemical parameters in any of the dose levels. No treatment related gross/histopathological lesions were observed. The present investigation demonstrated that the no observed adverse effect level was 2000 mg/kg body weight per day of hydroalcoholic extract of W. somnifera in rats and hence may be considered as non‐toxic. Copyright © 2012 John Wiley & Sons, Ltd.     

凉血化瘀方对肝衰竭大鼠细胞凋亡相关基因表达的调控作用

目的:应用功能分类基因芯片分析凉血化瘀方对急性肝衰竭大鼠细胞凋亡相关基因表达的影响,探讨其抗肝衰作用的分子机制。方法:腹腔注射GalN(500mg·kg^-1)+LPS(4.8μg·kg^-1),造成大鼠急性肝功能衰竭,应用美国SuperArray公司大鼠细胞凋亡Oligo基因芯片检测模型组和治疗组基因的差异表达。结果:模型组与正常组比较表达下调的基因有37条,表达上调的基因有10条;治疗组与模型组比较表达下调的基因有16条,其中诱导细胞凋亡基因有13条,表达上调的基因有12条,其中抗细胞凋亡基因7条。结论:凉血化瘀方能够调节细胞凋亡相关基因从而抑制肝细胞凋亡,主要是通过下调凋亡诱导相关基因的表达,同时也少量上调凋亡抑制基因的表达。     

肠易激综合征模型大鼠肺组织SP含量及基因表达变化研究

目的：检测肠易激综合征（IBS）模型大鼠肺组织P物质（SP）表达变化,探讨中医肺与大肠相表里脏腑相关理论的科学内涵。方法：采用慢性应激刺激与束缚相结合的方法建立IBS模型大鼠,复制成功后分别以免疫组化、酶联免疫吸附试验（ELISA）和逆转录多聚酶链反应（RT—PCR）等方法检测肺组织中的P物质（SP）表达。结果：与对照组比较IBS模型组大鼠的肺组织中SP含量以及基因表达明显增高（P〈0．01）。结论：IBS状态下大鼠肺组织中SP的含量及基因表达发生上调．提示肺与大肠通过神经一内分泌系统存在着内在联系。     

当归贝母苦参煎剂对慢性细菌性前列腺炎大鼠前列腺中超氧化物歧化酶和丙二醛的影响

目的观察当归贝母苦参煎剂对慢性细菌性前列腺炎(CBP)大鼠前列腺中超氧化物歧化酶(SOD)、丙二醛(MDA)含量的影响,探讨其可能的作用机制。方法雄性SD大鼠60只,随机分为空白对照组、模型组、西药对照组及当归贝母苦参煎剂高、中、低剂量组,每组10只。除空白对照组外,前列腺背叶注射大肠埃希菌制备CBP动物模型。测定各组大鼠前列腺中SOD、MDA的含量。结果与模型组比较,西药对照组、当归贝母苦参煎剂低剂量组大鼠前列腺中SOD明显升高,差异有统计学意义(P0.05),高剂量组大鼠SOD升高更为明显,差异有统计学意义(P0.01);西药对照组、当归贝母苦参煎剂各剂量组大鼠前列腺中MDA含量均明显降低,差异有统计学意义(P0.01)。结论当归贝母苦参煎剂对CBP所发挥的作用可能与升高前列腺组织中的SOD和降低MDA有关。     

回医烙灸疗法对脊髓损伤大鼠基因表达的调节机制

目的:观察回医烙灸疗法对脊髓损伤大鼠基因表达的调节机制,探讨调节的基因与神经组织保护、修复、生长和再生的关系。方法:采用Allen's法制备大鼠脊髓损伤模型,分为假手术组(A组),损伤组(B组),烙灸组(C组),应用表达谱基因芯片对脊髓损伤大鼠的基因表达分析,筛出先上调后下调及先下调后上调的基因,并用RT-PCR对其中8条基因表达水平验证。结果:电泳结果显示提取到高纯度的RNA,质量完好,无蛋白质及DNA残留。损伤组对比假手术组出现功能已有阐述差异基因1 146个,其中上调712个,下调434个。烙灸组对比B组出现功能已有阐述差异基因5 542个,其中上调3 006个,下调2 536个。模型组对比假手术组及烙灸组对比烙灸组基因差异表达取并集,然后从中挑选出样本中先上调后下调及先下调后上调的基因,共24个基因,其中先上调后下调的13个,先下调后上调的11个。抽取部分差异表达基因Olr1528、Trim72、Serpinb3a、GAP-43、Slc5a7、LOC288521、Vegp2、Cyp2d1进行RT-PCR验证,验证结果与芯片结果相符合。结论:回医烙灸疗法对脊髓损伤大鼠基因表达具有一定的调节作用,调节的基因与神经组织保护、修复、生长和再生密切相关。     

活血、益气方药对心衰大鼠血小板聚集影响的对比研究

本实验对比观察了活血注射液和益气注射液对心梗后左心衰动物血小板聚集率的影响，结果提示：二者对血小板聚集率均有明显改善，与模型组比较有显著性差异，与西药组相比无显著差异。益气注射液组对血小板聚集率的改善与活血注射液组无差异，说明单纯益气亦可起到活血的作用。     

加味菖蒲郁金汤对抑郁大鼠模型行为学的影响

目的:观察加味菖蒲郁金汤对抑郁大鼠模型行为学指标的影响。方法:经Open-Field法[1-3]评分测定评分相近的48只SD普通级大鼠,体质量为(200±20)g,雌雄各半,随机分为空白对照组(A)、模型组(B)、加味菖蒲郁金汤组(C)、氟西汀组(D)共4组,每组各12只。除对照组外,其他各组采用孤养和长期不可预见的慢性应激两种方法复制大鼠抑郁症模型[1]。以Open-Field法和糖水消耗实验对照研究慢性应激诱导的抑郁症模型大鼠行为学指标。结果:①与A组比较,各模型组大鼠在模型复制第21天Open-Field评分降低,糖水消耗量减少;②与B组比较,治疗组大鼠经治疗后Open-Field评分升高,糖水消耗量增加。结论:加味菖蒲郁金汤可改善孤养和长期不可预见的中等强度应激诱导的抑郁大鼠模型的抑郁行为。     

益肾降脂胶囊对慢性肾功能衰竭大鼠整体肾功能及血脂的影响

目的 观察益肾降脂胶囊对慢性肾功能衰竭(慢性肾衰)大鼠的保护作用，并探讨其治疗机制。方法 复制慢性肾衰大鼠模型，分别给大鼠灌胃益肾降脂胶囊(治疗组)和阳性对照药(阳性对照组)，90d后测大鼠肌酐(SCr)、尿素氮(BUN)及血脂、24h尿蛋白，取肾组织作病理检查；并与空白对照组比较。结果 与阳性对照组比较，治疗组BUN、SCr、24h尿蛋白及血脂降低，肾组织病理损害减轻。结论 益肾降脂胶囊具有降低血脂、改善肾功能、减轻肾间质炎症、促进受损组织恢复的作用。     

Inhibitory Effects of Multiple‐Dose Treatment with Baicalein on the Pharmacokinetics of Ciprofloxacin in Rats

Ciprofloxacin is used as a treatment for urinary and respiratory tract infections in clinical practice. Baicalein, a major flavonoid present in Scutellaria baicalensis, is a well‐known and potent antibacterial compound used in complementary and alternative medicine practices. The present study aimed to clarify the effects of multiple‐dose treatment with baicalein on the pharmacokinetics of ciprofloxacin in rats. Following the oral administration of baicalein (20, 40, or 80 mg/kg) for five consecutive days, the rats received an oral administration of ciprofloxacin (20 mg/kg). Blood samples were collected at specific time points, and the plasma concentrations of ciprofloxacin were determined by using high‐performance liquid chromatography. To evaluate the mechanisms underlying the interaction between baicalein and ciprofloxacin, a rhodamine 123 accumulation assay was performed in LS‐180 cells. A pharmacokinetic study revealed that multiple‐dose treatment with baicalein significantly decreased the peak serum concentration (C_max), area under the curve (AUC_{0 → 480 min}), and relative bioavailability (F_rel) of ciprofloxacin (p < 0.05). The rhodamine 123 accumulation assay revealed that treatment with baicalein for 48 h markedly reduced the intracellular accumulation of rhodamine 123. Taken together, these findings suggest that baicalein may result in the therapeutic failure of ciprofloxacin or other quinolone‐based antibiotics used for chemotherapy in clinical practice. Copyright © 2016 John Wiley & Sons, Ltd.     

毒素清对肺炎老龄大鼠小肠组织自由基和前列腺素代谢的影响

目的 :研究老年肺炎的小肠损伤病理生理特点与中药毒素清对其作用。方法 :复制肺炎双球菌肺炎模型 ,分为对照组、模型组、毒素清高低剂量组、头孢氨苄组。主要观察肺脏和小肠组织含水量 ,小肠组织前列腺素代谢产物 6 -Keto -PGF1α和TXB2 含量、NO和MDA含量及SOD活性。结果 :肺炎的肺组织损伤明显 ,肺脏和小肠组织含水量增高。模型组小肠组织 6 -Keto-PGF1α含量、SOD活性的降低和TXB2 、NO、MDA含量的增高较对照组显著。毒素清治疗后 ,SOD活性和 6 -Keto -PGF1α含量升高 ,MDA和NO含量下降。结论 :前列腺素及自由基介导的损伤参与肺炎的小肠组织损伤发生发展。毒素清对老龄大鼠肺炎的小肠损伤改善作用明显 ,其机制与其拮抗自由基损伤和调节前列腺素代谢平衡有关     

Effect of Ginkgo Biloba Extract on the Pharmacokinetics and Metabolism of Clopidogrel in Rats

Ginkgo biloba extract (GBE), a traditional herbal product used worldwide as both medicine and supplement, is often supplied with clopidogrel for the treatment of cerebrovascular diseases. The aim of the current study was to explore the effect of GBE on the metabolism and pharmacokinetics of clopidogrel. The in vitro study using rat liver microsomes revealed that GBE significantly induced the conversion of clopidogrel into its active metabolite. The effect of GBE on the pharmacokinetics of clopidogrel was also investigated in vivo. Compared to rats without GBE pretreatment, administration of 4 mg/kg, 20 mg/kg, and 100 mg/kg of GBE significantly decreased the C_max and the AUC_0–∞ of clopidogrel in a dose‐dependent manner. As expected, pretreatment of high dose GBE significantly increased the C_max and AUC_0–∞ of the clopidogrel active metabolite. However, no marked change was observed following medium and low dose of GBE, suggesting that the biotransformation of clopidogrel was altered differently by high dose of GBE. Our study suggested that the awareness of the potential herb–drug interactions between GBE and clopidogrel should be increased in clinical practice. Copyright © 2016 John Wiley & Sons, Ltd.