Effect of extract prepared from the roots of Paeonia lactiflora on endothelium-dependent relaxation and antioxidant enzyme activity in rats administered high-fat diet.

This study examined the effect of Paeoniae Radix (PR) on endothelial function and the activity of superoxide dismutase (SOD) of erythrocytes in rats administered a high-fat diet. Administration of the extract of PR increased the endothelium-dependent relaxation and the activities of SOD compared with high the cholesterol diet group significantly. Hypercholesterolaemia induced an increase of endothelial superoxide anion and endothelial dysfunction. Paeoniae Radix is suggested to have a protective effect on endothelial cells and their function.     

The drinking of a Salvia officinalis infusion improves liver antioxidant status in mice and rats

In this study, we evaluate the biosafety and bioactivity (antioxidant potential) of a traditional water infusion (tea) of common sage (Salvia officinalis L.) in vivo in mice and rats by quantification of plasma transaminase activities and liver glutathione-S-transferase (GST) and glutathione reductase (GR) enzyme activities. The replacement of water by sage tea for 14 days in the diet of rodents did not affect the body weight and food consumption and did not induce liver toxicity. On the other hand, a significant increase of liver GST activity was observed in rats (24%) and mice (10%) of sage drinking groups. The antioxidant potential of sage tea drinking was also studied in vitro in a model using rat hepatocytes in primary culture. The replacement of drinking water with sage tea in the rats used as hepatocyte donors resulted in an improvement of the antioxidant status of rat hepatocytes in primary culture, namely a significant increase in GSH content and GST activity after 4 h of culture. When these hepatocyte cultures were exposed to 0.75 or 1 mM of tert-butyl hydroperoxide (t-BHP) for 1 h, some protection against lipid peroxidation and GSH depletion was conferred by sage tea drinking. However, the cell death induced by t-BHP as shown by lactate dehydrogenase (LDH) leakage was not different from that observed in cultures from control animals. This study indicates that the compounds present in this sage preparation contain interesting bioactivities, which improve the liver antioxidant potential.     

筋骨片对骨性关节炎大鼠模型的影响

目的：观察筋骨片对骨性关节炎（OA）病变发生发展及其修复作用的影响。方法：将4％的木瓜蛋白酶与0.3 mol·L-1的半胱氨酸混悬液注入大鼠膝关节进行造模,分别给予蒸馏水、阳性对照药骨刺宁胶囊以及筋骨片药液灌胃治疗。连续给药4周后处死动物,进行各项指标的测定。结果：筋骨片对模型大鼠膝关节活动范围有明显的改善作用,各剂量组与模型组比较均有显著性差异;筋骨片各剂量组对模型大鼠膝关节滑膜厚度肿胀有抑制作用,能较好缓解OA的炎性症状,对制动后膝关节内滑膜及软骨的病理变化也有显著的改善作用。筋骨片各剂量组还能明显降低滑膜组织中的丙二醛（MDA）,减轻细胞损伤程度。结论：筋骨片减轻关节软骨的退变,明显抑制滑膜组织的炎症,减轻细胞的损伤程度,因而对OA具有一定的治疗效果。     

Hepatoprotective effect of flavonoid glycosides from Lespedeza cuneata against oxidative stress induced by tert-butyl hyperoxide

The aerial parts of Lespedeza cuneata G. Don, perennial legume native to Eastern Asia, have been used therapeutically in traditional Asian medicine to protect the function of liver, kidneys and lungs. However, little is known about the pharmaceutical effect of extracts from this plant. In the present study, the aerial parts of L. cuneata were used to prepare an ethanol extract, which was then tested for hepatoprotective effects against injury by tert-butyl hyperoxide (t-BHP). At a dose of 20 μg/mL, the ethanol extract significantly protected HepG2 cells against the cytotoxicity of t-BHP. Further fractionation of the extract with ethyl acetate allowed the isolation of five flavonoid compounds that were structurally identified by 1H and 13C NMR spectroscopy as isovitexin, hirsutrin, trifolin, avicularin and quercetin. Hirsutrin, avicularin and quercetin (10 μM) showed clear hepatoprotective activity against injury by t-BHP in HepG2 cells, whereas isovitexin and trifolin showed no protective effects. The observed hepatoprotective effect of the investigated compounds showed a high correlation with radical scavenging activity, which followed the structure-activity relationships of the flavonoid aglycones.     

左归丸通过线粒体途径抗叔丁基过氧化氢诱导的MC3T3-E1细胞凋亡

目的:观察左归丸对叔丁基过氧化氢诱导的MC3T3-E1细胞凋亡的保护效应,探讨其作用机制是否与其干预线粒体途径有关。方法:以成骨细胞MC3T3-E1为研究对象,制备左归丸含药血清,建立叔丁基过氧化氢(t-BHP)成骨细胞MC3T3-E1氧化应激模型,实验分为空白组,t-BHP组,左归丸+t-BHP组,补佳乐+t-BHP组。孵育24,48,72 h后,采用噻唑蓝(MTT)法检测左归丸含药血清对t-BHP诱导MC3T3-E1细胞存活的影响;孵育48 h后,采用吖啶橙溴乙啶(AO/EB)染色法检测细胞凋亡,采用Hoechst 33342染色法观察凋亡细胞核变化,采用罗丹明123荧光染色法观察线粒体膜电位的改变,采用蛋白质免疫印迹(Western blot)法分析线粒体蛋白家族B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax),半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平。结果:与空白组比较,t-BHP组显著抑制细胞增殖(P0.05,P0.01),细胞凋亡增加,线粒体膜电位降低,Bcl-2蛋白表达明显下调(P0.01);与t-BHP组比较,左归丸加t-BHP组促进暴露于t-BHP诱导的MC3T3-E1氧化损伤细胞的存活(P0.01),48 h者尤佳,逆转细胞凋亡情况,提高细胞线粒体膜电位,上调Bcl-2蛋白表达(P0.05),抑制Bax和Caspase-3蛋白表达(P0.01)。结论:左归丸含药血清能够抗t-BHP诱导的MC3T3-E1细胞凋亡,其作用机制可能与其干预线粒体途径有关。     

Hypericum androsaemum infusion increases tert-butyl hydroperoxide-induced mice hepatotoxicity in vivo

Increasing evidence regarding free radical generating agents indicates that the sustained production of high levels of reactive oxygen species (ROS) can cause hepatotoxicity. Being a short chain analog of lipid peroxide, tert-butyl hydroperoxide (t-BHP) is metabolized into free radical intermediates by cytochrome P450 in hepatocytes, which initiate lipid peroxidation, glutathione depletion and cell damage. The aim of the present study was to evaluate the putative protective effect of Hypericum androsaemum lyophilised infusion against t-BHP-induced mice hepatotoxicity in vivo, which has already been shown to be antioxidant in vitro. However, the results showed that the oral pretreatment with Hypericum androsaemum infusion (4, 20 and 100 mg/kg) for 4 days before a single intraperitoneal dose of t-BHP (1.8 mmol/kg) potentiated the t-BHP-induced hepatotoxicity. In fact, it was observed a potentiation in the depletion of total glutathione and reduced glutathione (GSH) contents and increase in oxidised glutathione (GSSG) level. Also the histopathological evaluation of the mice livers revealed that the infusion raised the incidence of liver lesions induced by t-BHP. These data do not corroborate any effect of Hypericum androsaemum infusion as hepatoprotector, but rather as a potentiator of hepatotoxicity in the present experimental conditions.     

筋骨片对兔骨性关节炎影响的实验研究

目的：观察筋骨片对骨性关节炎（OA）病变发生发展及其修复作用的影响。方法：用石膏绷带制动膝关节的方法给兔造模，给筋骨片药液灌胃治疗。连续给药4周，制动时间满6周时，将各组动物解除制动并停止给药。2周后进行各项指标的测定。结果：筋骨片对模型兔膝关节活动范围有明显的改善作用，各剂量组与模型组比较均有显著性差异；其中、高剂量的Ca／P比值也明显升高；同时筋骨片各剂量组对模型兔膝关节滑膜厚度肿胀有抑制作用，能较好缓解OA的炎性症状，对制动后膝关节内滑膜及软骨的病理变化也有显著的改善作用。筋骨片中、高剂量组还能明显增强滑膜组织SOD的活性，具有清除氧自由基的能力。结论：筋骨片减轻关节软骨的退变，明显抑制滑膜组织的炎症，提高SOD活力使OFR代谢恢复平衡，因而对OA具有一定的治疗效果。     

Protective activity of Hypericum androsaemum infusion against tert-butyl hydroperoxide-induced oxidative damage in isolated rat hepatocytes

Hypericum androsaemum L. (Guttiferae) is a medicinal plant with antioxidant activity. Increasing evidence regarding free radical generating agents suggests that hepatotoxic-related disorders may involve reactive oxygen species (ROS). The purpose of this study was to investigate the protective effect of Hypericum androsaemum infusion on isolated rat hepatocytes oxidative injury induced by tert-butyl hydroperoxide (t-BHP). The results showed that pretreatment of the cells with this infusion (16, 62 and 250 microg/ml) prevented the leakage of lactate dehydrogenase (LDH) and lipid peroxidation caused by a 30-min treatment with t-BHP (1mM). However, infusion-induced alterations on glutathione homeostasis were noticed, as it was observed by the increase in glutathione oxidised form (GSSG) and depletion in total glutathione levels, which indicates that plant-derived antioxidant extracts may not be considered a generalised way of treating pro-oxidant-related diseases.     

毛蕊异黄酮苷和芍药苷联用对氧糖剥夺再灌注HT22细胞PI3K/AKT信号通路的影响

目的观察并探讨毛蕊异黄酮苷和芍药苷联用对氧糖剥夺再灌注诱导的HT22细胞氧化应激损伤的作用及其机制。方法建立HT22细胞氧糖剥夺再灌注损伤模型,将HT22细胞分为正常对照组、模型组、给药组、尼莫地平组,应用CCK-8法检测细胞存活率,测定毛蕊异黄酮苷和芍药苷联用对HT22细胞活力的影响,检测细胞中乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量,并采用Western Blot法检测PI3K、AKT蛋白表达水平。结果与正常对照组相比,氧糖剥夺再灌注处理组的HT22细胞存活率明显降低(P<0.01),LDH活性及MDA的含量升高(P<0.05,P<0.01),SOD活力降低(P<0.01),PI3K、AKT蛋白表达明显下降(均P<0.01);与模型组相比,给药组能明显提高氧糖剥夺再灌注损伤后HT22细胞的存活率(P<0.01),降低LDH、MDA的含量(P<0.05,P<0.05),SOD活力提高(P<0.01),PI3K、AKT蛋白表达升高(均P<0.01)。结论毛蕊异黄酮苷和芍药苷联用对氧糖剥夺再灌注诱导的HT22细胞氧化应激损伤具有明显的保护作用,其作用机制可能通过调节PI3K/AKT信号通路实现。     

麦冬酒制前后对内皮细胞缺氧保护作用的比较研究

目的研究中药麦冬及其酒制炮制品对人脐静脉内皮细胞（EAHY926）缺氧损伤的保护作用。方法体外培养EAHY926,采用通入缺氧气体建立缺氧损伤模型。用麦冬提取液干预后,采用MTT比色法测定细胞活力、黄嘌呤氧化酶法测定细胞内超氧化物歧化酶活性、硫代巴比妥酸显色法测定丙二醛含量、硝酸还原酶法测定一氧化氮含量。结果组间比较采用单因素方差分析,两两之间用SNK检验。与模型组相比,川麦冬生品提取液组、酒制川麦冬提取液组、杭麦冬生品提取液组、酒制杭麦冬提取液组均能显著提高细胞活力（P〈0.05）,并且均显著提高缺氧后细胞超氧化物歧化酶活性（P〈0.05）及降低丙二醛、一氧化氮的含量（P〈0.05）;酒制川麦冬提取液组和酒制杭麦冬提取液组效果最佳,与空白对照组比有非常显著性差异（P〈0.01）。结论麦冬酒制后更能显著拮抗缺氧对EAHY926的损伤。     

合欢汤治疗冠心病临床研究

将５９例冠心病患者分为中药合欢汤组（３７例）和心痛定对照组（２２例）进行自身治疗前后对照和组间比较。结果：合欢汤组缓解心绞痛显效率和总有效率分别为７５．６８％与９１．８９％；治疗后血清超氧化物歧化酶（ＳＯＤ）、全血谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）活性显著高于治疗前（Ｐ＜０．０１）；且ＳＯＤ活性、ＧＳＨ－ＰＸ／ＬＰＯ比值高于对照组（Ｐ＜０．０１与Ｐ＜０．０５）、ＬＰＯ含量低于对照组（Ｐ＜０．０５）。提示合欢汤具有抗心绞痛、升高抗氧化酶活性、降低脂质过氧化作用。     

川断总皂苷对大鼠局灶性脑缺血损伤保护作用的实验研究

目的 研究川断总皂苷抗脑缺血的作用.方法 将40只大鼠随机分成为正常组、模型组、川断总皂苷低剂量组(10mg/kg)和高剂量组(50mg/kg).采用插线法阻塞大鼠大脑中动脉,制造大鼠局灶性脑缺血模型.作脑匀浆,测定脑匀浆中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)含量等指标.结果 川断皂苷低、高剂量组均能明显减少脑缺血大鼠的脑梗死面积,提高脑缺血脑中SOD、GSH-Px活性,并降低其MDA的含量.结论 川断总皂苷对大鼠局灶性脑缺血损伤有明显的保护作用.     

琥珀调控颈总动脉结扎脑缺血大鼠海马区神经元凋亡改善大鼠学习与记忆能力的效用部位与作用机制研究＊

目的 琥珀调控海马区神经元凋亡改善颈总动脉结扎脑缺血大鼠学习与记忆能力的效用部位与作用机制研究。方法 建立大鼠颈总动脉结扎脑缺血动物模型，给以模型动物琥珀系列溶剂提取部位，通过水迷宫实验评价琥珀提取部位对模型动物学习与记忆能力的影响。采用TUNEL染色法与蛋白印迹法分别评价琥珀提取部位对模型动物海马区细胞凋亡以及Caspase 3、Caspase 9、Bcl-2等凋亡相关蛋白表达的影响。采用小鼠海马永生化细胞系评价琥珀提取部位对抗糖氧剥夺神经元损伤的作用。结果 琥珀乙酸乙酯提取部位显著改善模型大鼠学习与记忆能力，抑制海马区细胞凋亡。该提取部位还可显著提高糖氧剥夺状态下HT22细胞的存活率，并下调细胞中Caspase 3、Caspase 9和Bcl-2等凋亡通路蛋白表达水平。结论 乙酸乙酯提取部位是琥珀药材改善颈总动脉结扎脑缺血大鼠的学习与记忆能力效用部位，抑制海马区细胞凋亡是其作用机制。     

甘草蜜炙前后对小鼠免疫调节的影响

目的 研究甘草炮制前后对免疫低下小鼠免疫调节作用的影响。方法 小鼠腹腔注射环磷酰胺制备免疫低下模型,给予生甘草、清炒甘草、蜜炙甘草的提取液,检测免疫功能学评价指标,包括免疫器官指数、半数溶血值、溶血空斑数、脾淋巴细胞增值率、自然杀伤（NK）细胞杀伤率、耳肿胀度、碳廓清指数、腹腔巨噬细胞吞噬率及吞噬指数。结果 与对照组小鼠比较,模型组小鼠各检测指标明显降低（P<0.05,P<0.01）;与模型组小鼠比较,各给药组（生甘草组、清炒甘草组、蜜炙甘草组）小鼠各检测指标均能增强。其中,仅蜜炙甘草组能明显增强各检测指标（P<0.05,P<0.01）;与生甘草组小鼠相比,清炒甘草组能明显增强血清溶血素和抗体生成细胞、脾淋巴细胞增殖率和NK细胞活性（P<0.05,P<0.01）,但对胸腺指数、脾脏指数、腹腔巨噬细胞吞噬率和吞噬指数、DTH、碳廓清功能无明显影响,蜜炙甘草组能明显增强除胸腺指数的各项评价指标（P<0.05,P<0.01）,但对胸腺指数无明显影响。结论 甘草蜜炙前后对小鼠非特异性免疫、体液免疫及细胞免疫功能均有促进作用,且蜜炙后作用更强,为蜜炙甘草机制研究提供参考。     

Polygalae radix extract protects cultured rat granule cells against damage induced by NMDA

Polygalae Radix (PR) from Polygala tenuifolia (Polygalaceae) is traditionally used in China and Korea, as this herb has a sedative, anti-inflammatory and antibacterial agent. To extend our understanding of the pharmacological actions of PR in the CNS on the basis of its CNS inhibitory effect, the present study examined whether PR has the neuroprotective action against N-methyl-D-aspartate (NMDA)-induced cell death in primarily cultured rat cerebellar granule neurons. PR, over a concentration range of 0.05 to 5 microg/ml, inhibited NMDA (1 mM)-induced neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. PR (0.5 microg/ml) inhibited glutamate release into medium induced by NMDA (1 mM), which was measured by HPLC. Pre-treatment of PR (0.5 microg/ml) inhibited NMDA (1 mM)-induced elevation of intracellular Ca2+ concentration ([Ca2+]i), which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that PR prevents NMDA-induced neuronal cell damage in vitro.     

近十年葛根防治骨质疏松症的研究进展

葛根是一种药食同源植物,含有大量异黄酮。实验研究表明葛根及相关提取物在体内外实验中有增加骨量、改善骨代谢的功能。临床研究发现葛根能改善骨质疏松症病人的骨代谢生化指标,并改善患者的临床症状,一些防治骨质疏松症的中药复方也开始加入葛根,说明葛根治疗骨质疏松症已得到认可并应用于临床。本文对近十年来葛根防治骨质疏松症的研究进展作一综述,并对未来做出展望。     

Protection of brain cells against AMPA-induced damage by Asiasari Radix extracts

We determined whether Asiasari Radix (AR) extracts have protective actions in brain cells. Methanol extracts of Asiasari Radix (fraction 1) have significant inhibitory effects on the AMPA-induced rat cortical depolarization in the grease gap assay. In differentiated PC12 cells, it almost completely protected against AMPA-induced cell death. In addition, it had some protective actions in C6 glial cells death induced by AMPA. The methanol extracts (fraction 1) of AR were subsequently fractionated into chloroform-(fraction 2), chloroform/methanol-(3:1) (fraction 3), methanol-soluble (fraction 4) and methanol-insoluble, water-soluble fractions (fraction 5). Among these, fraction 4 had the strongest inhibitory effects against AMPA-induced cell death in the PC 12 cells and also dramatically inhibited AMPA-induced depolarization of rat brain cortex in the grease gap assay. Interestingly, fraction 4 blocked the Zn-induced oxidative damages in C6 glial cells.     

麋鹿角调控慢性压力应激小鼠神经营养因子和压力应激因子抗抑郁作用研究＊

目的 考察麋鹿角提取物调控神经营养因子和压力因子抗抑郁的作用机制。方法 制备麋鹿角提取物，采用小鼠慢性应激压力（chronic unpredictable mild stress,CUMS）模型，通过糖水偏嗜实验、悬尾实验和强迫游泳实验评价麋鹿角提取物的抗抑郁功效。采用ELISA法测定受试动物海马区中神经生长因子（Nerve growth factor,NGF）、脑源性神经营养因子（brain-derived neurotrophic factor,BDNF）的含量。采用ELISA测定受试动物血清中相应压力因子：促肾上腺皮质激素释放激素（corticotropinreleasing factor,CRF）、促肾上腺皮质激素（adreno-cortico-tropic-hormone,ACTH）、皮质醇的含量。采用大鼠星形胶质瘤细胞（C6）评价麋鹿角提取物调控神经营养因子系统的作用，并采用小鼠海马永生化细胞系（HT22）评价其对抗压力因子损伤的作用。结果 麋鹿角提取物能显著上调模型小鼠糖水偏嗜率，缩短小鼠悬尾和强迫游泳不动时间，以麋鹿角水提物的效用最强，该提取物能通过Erk信号通路，显著上调C6细胞中的NGF和BDNF的表达。此外，在皮质醇（Cortisol）损伤HT22的体外模型中，麋鹿角水提物具有显著的细胞保护作用，效果优于醇提物。结论 麋鹿角对神经营养因子的调控和压力因子的调控是其抗抑郁的重要机制。     

银翘散及其拆方对流感病毒感染自然杀伤细胞活性的影响及转录组的比较分析

目的研究银翘散及其拆方(君药、臣药、佐药、使药)对流感病毒感染自然杀伤细胞活性及转录组的影响。方法以小鼠流感病毒PR8(A/Puerto Rico/8/1934,H1N1型)感染ICR小鼠制备肺炎模型,分为对照组、模型组和给药组,每组10只;银翘散、君药(金银花、连翘)、臣药(薄荷、牛蒡子、淡豆豉、荆芥穗)、佐药(桔梗、芦苇根、淡竹叶)和使药(甘草)、阳性药物利巴韦林分别ig模型小鼠5 d;第5天检测小鼠体质量、肺质量,计算肺质量抑制率;固定肺脏,观察病理学变化(HE染色)。以PR8病毒感染C57BL/6J小鼠制备肺炎模型,分为对照组、模型组和给药组,每组3只,上述药物分别ig5d,第5天摘取肺脏,制备白细胞悬液,采用流式细胞术检测自然杀伤(naturekiller,NK)细胞表面激活性受体Ly-49D和Ly-49H的含量,以及细胞毒性受体NKp46的含量。为了进一步探究药物与病毒感染NK细胞之间的互作关系,建立人流感病毒HK8(A/HongKong/8/68,H3N2型)感染人自然杀伤细胞系NK-92MI模型,比较银翘散及臣药对病毒感染NK细胞转录组作用的异同。结果银翘散及其拆方对PR8病毒感染引起的小鼠肺炎均有一定的抑制作用,抑制作用由强到弱为臣药>银翘散>使药>君药>佐药,其中臣药和银翘散组具有统计学差异(P<0.05)。银翘散及其拆方均能增加肺脏NK细胞激活性受体Ly-49D和Ly-49H的含量,其促进作用由强到弱为臣药>使药>银翘散>君药>佐药,其中银翘散、君药、臣药和使药组均具有统计学差异(P<0.05、0.01)。结合上述2个指标,臣药表现出与银翘散相似的药效特征,且略优于银翘散。对于PR8病毒感染小鼠肺脏NK细胞毒性受体NKp46的含量,臣药也表现出良好的诱导表达作用。HE染色显示,经臣药治疗的PR8病毒感染小鼠,其肺脏结构完好,肺泡和支气管腔内未出现炎症细胞和红细胞浸润。NK细胞转录组分析显示,臣药对于HK8病毒感染的NK细胞的作用位点,不仅囊括了银翘散的作用位点,而且比银翘散更为广泛。结论对于流感病毒感染小鼠,银翘散的拆方臣药表现出与银翘散全方相似的药效,且略优于银翘散,这可能与臣药对NK细胞活性的调节作用比银翘散更广泛有关。