Analgesic and anti-inflammatory activities of the ethanol extract of the leaves of Helianthus Annus in Wistar rats

Objective:To investigate the anti-inflammatory and analgesic effects of the ethanol extract of leaves of Helianthus annus L.(H.annus) in acclimatized Wistar rats.Methods:It was undertaken using the albumin induced paw edema model of inflammation as well as both the hotplate and tail immersion analgesic test methods.Doses of the extract tested in experimental rats were 0.5 g/kg,2 g/kg and 4 g/kg while negative and positive control rats received distilled water and indomethacin respectively.Results:It was shown that treatment with the tested doses of the extract effectively inhibited paw edema induced by egg albumin.This effect was comparable if not better than the observations made in rats treated with 10 mg/kg of indomethacin orally.Treatment with the extract was also observed to have significantly increased the mean tolerance time of rats to thermal noxious stimuli compared to control animals that had distilled water and appeared to be more effective than 10 mg/kg of indomethacin treatment.Conclusions: These observations confirmed the presence of a strong anti-inflammatory and anti-noiciceptive activity in the ethanol extract of the leaves of H.annus and therefore validated the folkloric use of the leaves of this plant in treatment of pro-inflammatory,post traumatic situations.

Antidiabetic and anti-lipemic effects of Cassia siamea leaves extract in streptozotocin induced diabetic rats

Objective:To investigate the antidiabetic and anti-lipemic effects of Cassia siamea methanolic leaves extract.Methods:The antidiabetic study was performed by measuring blood glucose level with elegance glucometer at weekly intervals i.e.0,7,14 and 21 in normal and streptozotocin induced diabetic rats.Total cholesterol,triglyceride and HDL-cholesterol were determined in normal and streptozotocin induces diabetic rats by autoanalyser.Glibenclamide was used as a reference drug at a dose of 10 mg/kg.Results:After the oral administration of extracts at doses of 250 mg/kg and 500 mg/kg for three weeks,blood glucose levels and body weights were significantly improved(P【0.01).Daily oral treatment with the extract also resulted in significantly reduction of serum cholesterol and triglycerides.HDL-cholesterol level was found to be improved to(P【0.01).Conclusions:The Cassia siamea leaf extract is useful in controlling blood glucose level as well as improving lipid metabolism and body weight in rats with induced diabetes.

Effect of Boschniakia rossica on expression of GSTP,p53 and p21^(ras)proteins in early stage of chemical hepatocarcinogenesis and its anti-inflammatory activities in rats

AIM To investigate the effect of Boschniakiarossica（BR）extract on expression of GST-P,p53 and p21rasproteins in early stage of chemicalhepatocarcinogenesis in rats and its anti-inflammatory activities.METHODS The expression of tumor marker-placental form glutathione S-transferase（GST-P）,p53 and p21rasproteins were investigated byimmunohistochemical techniques and ABCmethod.Anti-inflammatory activities of BR werestudied by xylene and croton oil-induced mouseear edema,carrageenin,histamine and hotscald-induced rat pow edema,adjuvant-inducedrat arthritis and cotton pellet-induced mousegranuloma formation methods.RESULTS The 500 mg/kg of BR-H2O extractfractionated from BR-Methanol extract hadinhibitory effect on the formation of DEN-inducedGST-P-positive foci in rat liver（GST-P stainingwas 78% positive in DEN+AAF group vs 20%positive in DEN+AAF+BR group,P【0.05）andthe expression of mutant p53 and p21rasproteinwas lower than that of hepatic preneoplasticlesions（33% and 22% positive respectively inDEN+AAF group vs negative in DEN+AAF+BRgroup）.Both CH2Cl2 and H2O extracts from BRhad anti-inflamatory effect in xylene and crotonoil-induced mouse ear edema（inhibitory rateswere 26%-29% and 35%-59%,respectively）. BR-H2O extract exhibited inhibitory effect incarrageenin,histamine and hot scald-inducedhind paw edema and adjuvant-induced arthritis inrats and cotton pellet-induced granulomaformation in mice.CONCLUSION BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in early stage of rat chemical hepato-carcinogenesis. Both CH2CI2 and H2O extracts from BR exerted anti-inflammatory effect in rats and mice.

Anti-Obesity, Anti-Atherosclerotic and Anti-Oxidant Effects of Pu-Erh Tea on a High Fat Diet-Induced Obese Rat Model

Pu-erh tea, a traditional Chinese beverage, has been believed to have many benefits to human health and without side effects. In this study, we systematically analyzed the main active components of Pu-erh tea and investigated its anti-obesity, anti-atherosclerotic and anti-oxidant effects using an obese rat model. Obesity was induced by feeding a high-fat diet and subsequently the experimental obese mice were fed with high-fat diet supplemented with low (2.5%), medium (5%) or high (7.5%) doses of Pu-erh tea powder for 6 weeks respectively. As result, the body weight gain of the rats was decreased by medium and high doses of Pu-erh tea treatments. Furthermore, the levels of serum total cholesterol (TC), triglyceride (TG) and atherosclerosis index (AI) were significantly lowered by Pu-erh tea compared to the control group. Conversely, high density lipoproteincholesterol (HDL-C) level of the rats was significantly elevated by Pu-erh tea treatments. In addition, Pu-erh tea treatments increased the activities of anti-oxidative enzymes such as superoxide dismutase (SOD) and glutathione peroxides (GSH-Px), whereas reduced the level of lipid peroxidation product malondialdehyde (MDA) in obese rats. Collectively, our find-ings revealed that Pu-erh tea exerts comprehensive benefits in anti-obesity, anti-atherosclerotic and anti-oxidant effects, therefore can be used as a promising functional food in obesity management.

Antifertility effect of chronically administered Tabernaemontana divaricata leaf extract on male rats

Objective:To investigate the antifertility effect of chronically administered Tabernaemontana divarkala(T.divaricata) leaf extract on male rats.Methods:The effect of 50%ethanol extract of T.divaricata leaves on reproduction was studied on male rats.The study was divided into four groups of five animals each.The first groups(1) received vehicle alone to serve as control. The second,third and fourth groups(Ⅱ,ⅡandⅣ) of animals were administered the leaf extract daily at 50 mg/kg body weight,p.o.,100 nig/kg body weight,p.o.,and 200 mg/kg body weight, p.o.,respectively,for a period of 60 days.Results:Significant decreases in the weight of testes, epididymis,seminal vesicle and ventral prostate were observed.A dose related reduction in the testicular sperm count,epididymal sperm count and motility,number of fertile male,ratio between delivered and inseminated females and numbers of pups were observed.The testis showed a clear correlation between the dose and severity of lesions of seminiferous epithelium. In general,the seminiferous tubules appear reduced in size with a frequently filled eosinophilic material.Spermatogenesis arrested at the secondary spermatocyte stage.Pachytene spermatocytes were undergoing degeneration.Disorganisation and sloughing of immature germ cell were visible. Leydinf cells were atrophied.No morphological changes were observed in Sertoli cells.Significant reduction in serum concentration of luteinizing hormone and testosterone were observed.No distinct change in serum FSH concentration was recorded.The final body weights of all groups were elevated markedly.No alterations were recorded in any hematologiocal parameters. Conclusions:It is concluded that the 50%ethanol extract of T.divaricata leaf produced dose related effect on male reproduction without altering general bodv metabolism.

Preparation and Identification of Rat Monoclonal Anti-idiotope Antibody to the Antibody against Erythrocytic Stages of Plasmodium Falciparum

A hybridoma cell line secreting monoclonal antibody to idiotope of the monoclonalantibody 94D1 specific for asexual erythrocytic stages of Plasmodium falciparum was established byfusion of SP2/0 mouse myeloma cells with spleen cells of Wistar rats immunized with monoclonalIgG of 94D1 purified from ascitic fluid of BALB/C mouse by affinity chromatography on ProteinA- Sepharose CL- 4B. Specificity of the anti - idiotope antibody (anti - Id), 41RF5, was deter-mined with indirect enzyme-linked immunosorbent assay (ELISA). The results showedthat 41RF5 reacted only with 94D1 IgG2b, not with normal mouse IgG and other seven mousemonoclonal antibodies - five specific for erythrocytic stages of P. falciparum, one for erythrocyticstages of P. inui, and one for Dengue fever virus type III, which indicates that 41RF5 does recognizespecifically the idiotope of 94D1 monoclonal antibody. In 41RF5 heterohybrid culture supernatant,anti-Id titre measured by ELISA was above 1:1 280. Up to the present, the heterohybrid cell linehas been cultured stably for 16 months.

Aliskiren Augments the Activities of Anti-Oxidant Enzymes in Liver Homogenates of DOCA Salt-Induced Hypertensive Rats

Hypertension is a serious problem that is recently thought to be associated with damaging effects on target organs partially via oxidative stress. On the other hand, there is accumulating literature describing some sort of therapeutic interaction between antioxidant enzymes in vital organs and hypertension. Therefore, the aim of this study is to investigate the possible effect of a direct renin inhibitor, aliskiren, used in treatment of hypertension via renin-angiotensin-aldosterone system (RAAS), on selected anti-oxidant enzymes in hepatic homogenates in DOCA salt-induced hypertesnive albino rats. Thirty male wister albino rats were assigned randomly into 3 groups (n = 10/ group). Group 1 received no treatement and serves as control. Group 2 received 0.5% carboxymethylcellulose sodium ip as a solvent of aliskiren, as a direct renin inhibitor (DRI). Group 3 received aliskiren 100 mg/kg/day ip for 4 weeks through gastric tube. Systolic blood pressure (SBP) was measured every week and its mean was recorded at the end of the study. Superoxide dismutase (SOD) enzyme in RBCs lysates, activities of catalase (CAT) and glutathione peroxidase enzymes and thiobarbituric acid reactive substance (TBARS), as a marker of lipid peroxidation, in hepatic homogenates were measured at the end of the study. DRI produced a marked reduction in mean SBP of hypertensive rats. It also significantly (p < 0.05) increased the activities of measured anti-oxidant enzymes while it significantly (p < 0.05) reduced TBARS in liver homogenates. These results indicated that renin possesses an oxidative effect in the liver in hypertensive rats. Aliskiren, in addition to its powerful anti-hypertensive effect, it could induce a great anti-oxidant effect in liver homogenates of DOCA salt-hypertensive rats.

Safety Assessment and Potential Anti-Inflammatory Effect of Ethanolic Extract of <i>Syzygium aromaticum</i>in Albino Rats

The present study was undertaken to investigate the effect of the ethanolic extract of Syzygium aromaticum to albino rats. Forty eight Albino rats were employed to test the safety and the anti-inflammatory effect of the extract. Safety of the extract was examined on experimental animal’s model at three dose levels of the extract orally in daily doses for three weeks. Effects of S. aromaticum on rats revealed no significant effect on biochemical or haematological parameters. The anti-inflammatory effect of the extract was tested in four equal groups;groups 1 and 2 were treated with 250 and 500 mg/kg of the extract, respectively, group 3 was treated with indomethacine and group 4 was the untreated control. Carrageenan was used as an acute form inducer of inflammation. Indomethacine, the non-steroidal anti-inflammatory drugs (NSAIDs), was used as a reference compound. Oedema size was monitored at the 1st, 2nd, 4th, 6th and after 24 hours. The ethanolic extract of S. aromaticum showed significant (P 0.001) decreased in the oedema size at efficacy rates of 79.41%, 82.39% and 63.92% for the dose, 500 mg/kg body weight at the 2nd, 4th and 6th hour respectively higher than that produced by indomethacine.

Phytochemical Screening, Toxicity, Analgesic and Anti-Pyretic Studies of Aqueous Leaf Extract of <i>Plectranthus barbatus</i>[Andrews. Engl.] in Rats

Plectranthus barbatus is a popular tropical perennial plant with a wide variety of traditional medicinal uses in tropical Africa, Hindu, Ayurvedic and traditional medicines of Brazil and China. The whole plant and the leaves have many folkloric uses for diverse ailments including pain, heart disease, convulsions, coughs and colds, asthma, bronchitis and tonsillitis among others. This study investigated the phytochemical components, acute toxicity, analgesic and anti-pyretic activities of the aqueous leaf extract of Plectranthus barbatus locally known as Ekizeera in Uganda. The plant leaves were authenticated, collected and decoction was done according to local method. Phytochemical screening was conducted using methods outlined by Trease and Evans and Harborne to determine the components of the extract. Acute toxicity tests were conducted in rats using modified Lorke’s method to determine the safety of the plant material. Analgesic studies were carried out using both a mechanical method (thermally induced pain by tail-flick) and a chemical method (formalin induced pain) in rats by administering extracts orally at 100, 200 and 400 mg/kg of body weight. The method of Al-Ghamdi, modified for local laboratory setting by Adzu was adopted and used for anti-pyretic test. Decoction yielded 9.9% extract. Phytochemical screening confirmed presence of saponins, tannins, alkaloids, terpenoids and essential oils. Acute toxicity tests revealed no deaths in rats after oral treatment with up to 10,000 mg/kg of extract. Tail-flick test was non-significant (p > 0.05) while formalin-induced pain test demonstrated significant activity (p -tests. Anti-pyretic activity was non-significant (p > 0.05) with student t-test. These results suggest that the aqueous leaf extract of Plectranthus barbatus contains specific phytochemicals, has a potent dose dependent analgesic activity, no anti-pyretic activity and can be regarded as a safe medicinal plant to use traditionally, which might further be developed for conventional medical practice.

The expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells in asthmatic rats and the effect of anti-CD40L McAb on Th1 and Th2 cytokines

Objective： To investigate the expression of CD40 and CD40 ligand （CD4OL） on the surface of peripheral blood mononuclear cells（PBMCs） in asthmatic rats and the effect of anti-CD40L McAb on cytokines of PBMCs. Methods： Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs in asthmatic rats. After the PBMCs was treated with anti-CD40L McAb, ELISA was used to detect the IL-4 and IFN-γ levels of culture supernatants. Results： Compared with the normal control group, the expression of CD40 and CD40L of PBMCs in asthmatic rats increased （P 〈 0.05）. Compared with the untreated group, the level of IL-γ and the ratio of IL-4/IFN-γ decreased after the PBMCs was treated with anti-CD40L McAb（P 〈 0.05）. Conclusion： The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats was up-regulated. Anti-CD40L McAb can rectify the imbalance of Th1 and Th2 cytokines.

乳痛软坚片抗炎镇痛及调节免疫药效作用的实验研究

目的:探讨乳痛软坚片抗炎、镇痛及调节免疫的药效作用。方法:通过小鼠扭体实验、小鼠热板法实验,采用醋酸扭体及热板致痛模型,以扭体次数、痛域值为检测指标,观察乳痛软坚片的镇痛作用;通过大鼠肉芽肿实验、大鼠足肿胀实验,采用棉球植入诱导大鼠肉芽肿增生、角叉菜胶致炎模型,以肉芽重量、足跖肿胀度为检测指标,观察乳痛软坚片抗炎作用;通过小鼠血清溶血素实验、小鼠碳粒廓清实验,采用腹腔注射5%氯化钠溶液鸡红细胞混悬液、小鼠尾静脉注射20%印度墨汁(0.1 mL/10 g)制造小鼠免疫抑制模型,以血清溶血素、廓清指数(K)、吞噬指数(α)为检测指标,观察乳痛软坚片的调节免疫作用。结果:扭体实验中,乳痛软坚片高剂量组小鼠扭体潜伏期长于模型组,扭体次数少于模型组(P<0.01或P<0.05);乳痛软坚片中剂量组小鼠扭体次数少于模型组(P<0.05)。热板法实验中,乳痛软坚片组高、中剂量组小鼠给药后1、2、3 h痛域值高于模型组(P<0.01或P<0.05)。大鼠肉芽肿实验中,乳痛软坚片组高、中、低剂量组大鼠肉芽质量均低于模型组(P<0.01或P<0.05)。大鼠足肿胀实验中,乳痛软坚片高剂量组大鼠在致炎后1、3、4 h足跖肿胀度低于模型组(P<0.05),乳痛软坚片中剂量组大鼠在致炎后3、4 h足跖肿胀度低于模型组(P<0.05)。小鼠血清溶血素实验中,乳痛软坚片高剂量组小鼠溶血素高于模型组(P<0.01)。小鼠碳粒廓清实验中,乳痛软坚片高、中、低剂量组小鼠廓清指数、吞噬指数均高于模型组,差异均有统计学意义(P<0.01或P<0.05)。结论:乳痛软坚片具有良好的抗炎、镇痛及调节免疫的药理作用。

基于TLR4/NF-κB和Nrf2/p62通路研究三臣散对幼鼠肺部炎症的抗炎及抗氧化作用机制

目的:基于Toll样受体4/核因子κB(TLR4/NF-κB)和核因子E2相关因子2/泛素结合蛋白p62(Nrf2/p62)通路研究三臣散(SCP)对脂多糖(LPS)诱导的幼鼠肺部炎症的抗炎及抗氧化作用机制。方法:将76只雄性Wistar幼鼠随机分为空白组(n=12)和造模组(n=64)。造模组幼鼠通过LPS经口气管滴注诱导肺部炎症,将造模后幼鼠随机分为三臣散高剂量组(HSCP组)、三臣散低剂量组(LSCP组)、地塞米松组(DEX组)和模型组(Model组),每组16只。HSCP组大鼠给予0.10 g/mL SCP灌胃,LSCP组给予0.05 g/mL SCP灌胃,DEX组给予0.025 g/mL地塞米松灌胃,Model组和空白组给予1 mL/100 g羧甲基纤维素钠(CMC-Na)溶液灌胃,2次/d。分别于第2、4天末次给药后24 h取材(n=8)。取材后先行肺部病理切片检测造模是否成功,各组取12只(第2、4天各6只)造模成功幼鼠进行后续指标检测。观察幼鼠的一般情况、体质量,检测凝血四项、血常规,HE染色观察肺组织病理情况,酶联免疫吸附试验(ELISA)检测下丘脑发热中枢介质和支气管肺泡灌洗液(BALF)中炎症因子的含量,检测肺组织超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,免疫印迹法检测肺组织中TLR4、IκBα、p65和Nrf2、p62蛋白表达水平。结果:与空白组比较,模型组幼鼠肺部在第2天出现大量炎症细胞浸润,肺泡结构破坏,肺泡壁出现透明膜,肺泡壁增厚、融合,支气管有片状渗出物;第4天肺泡壁增厚加重,肺泡数量减少,肺间质纤维化。与模型组比较,HSCP组、LSCP组、DEX组幼鼠肺部整体损伤程度更低,其中HSCP组最低。第2天,模型组幼鼠中性粒细胞数、前列腺素E(PGE)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、TLR4表达量、MDA含量均高于空白组(P<0.05),环磷酸腺苷(cAMP)、转化生长因子-β(TGF-β)均低于空白组(P<0.05);第4天,模型组幼鼠中性粒细胞数、PGE、IL-6、白介素-4(IL-4)、白介素-10(IL-10)、TNF-α、MDA含量均高于空白组(P<0.05),cAMP、TGF-β、IκBα、SOD水平均低于空白组(P<0.05)。第2天,HSCP组幼鼠cAMP、TGF-β高于模型组、LSCP组和DEX组(P<0.01或P<0.05),PGE、IL-6、TNF-α、IL-4、MDA含量低于模型组、LSCP组和DEX组(P<0.01或P<0.05);HSCP组幼鼠IL-10高于模型组(P<0.01),TLR4表达量高于LSCP组、DEX组(P<0.05);DEX组幼鼠TLR4表达量低于模型组(P<0.05)。第4天,HSCP组、LSCP组和DEX组幼鼠cAMP、IL-10、TGF-β高于模型组(P<0.05或P<0.05),中性粒细胞、PGE、IL-6、TNF-α、IL-4、MDA含量低于模型组(P<0.05或P<0.01)。各组幼鼠第4天TLR4、IκBα蛋白相对表达量比较,差异均无统计学意义(P>0.05)。结论:SCP可以减轻LPS诱导的幼鼠轻型肺部炎症,其作用机制可能是通过调控TLR4/NF-κB通路下游的炎症因子来抑制炎症反应,以及激活Nrf2/p62通路调控与氧化应激相关的因子来抑制氧化应激反应。

小儿风热清合剂抗炎解热镇痛药理作用研究

目的:探索小儿风热清合剂的抗炎解热镇痛药理作用。方法:采用二甲苯致小鼠耳肿胀模型,测定小鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)的含量,评价小儿风热清合剂的抗炎药理作用;采用干酵母致大鼠发热模型评价小儿风热清合剂的解热药理作用;采用冰醋酸致痛扭体法和热板法评价小儿风热清合剂的镇痛药理作用。结果:阳性对照组和小儿风热清低、中、高剂量组二甲苯致小鼠耳廓肿胀度低于模型组,差异均有统计学意义(P<0.01或P<0.05);小儿风热清中、高剂量组小鼠血清TNF-α含量均低于模型组,差异均有统计学意义(P<0.01);小儿风热清高剂量组小鼠血清IL-6含量低于模型组(P<0.05)。小儿风热清高剂量组小鼠脾脏指数低于模型组(P<0.05)。小儿风热清低、中、高剂量组大鼠体温低于模型组,且呈现剂量依赖性,差异均有统计学意义(P<0.01或P<0.05)。小儿风热清中、高剂量组小鼠扭体反应次数低于模型组(P<0.05);小儿风热清合剂可提高热板实验小鼠的痛阈值,随着小儿风热清合剂剂量增加,小鼠痛阈值呈正相关提高,差异有统计学意义(P<0.05)。且其镇痛效果在60 min内达到峰值。结论:小儿风热清合剂具有良好的抗炎解热镇痛药理作用。

生脉饮加味对运动性疲劳模型大鼠心功能指标的影响

为了探究生脉饮加味对运动性疲劳模型大鼠心功能指标的影响,本试验将50只雄性Wistar健康大鼠随机分为5个组,每组10只。空白组灌服生理盐水1 mL/(kg·bw·d),正常饲养。模型组灌服生理盐水1 mL/(kg·bw·d),低、中和高剂量组按照0.25、0.5和1 g/(kg·bw·d)灌服同等体积的生脉饮加味,进行7周速度递增跑台运动,结束后1 d进行力竭运动,记录力竭时间,心脏采血并分离血清,测定心肌酶谱、血气和抗氧化指标。结果显示,中剂量组大鼠力竭时间明显长于模型组(P<0.05)。心肌酶谱检测结果显示,与空白组相比,模型组α-羟丁酸脱氢酶(α-HBDH)、肌酸激酶(CK)、乳酸脱氢酶(LDH)和谷草转氨酶(AST)的酶活性均显著提高(P<0.05),而各给药组无显著差异(P>0.05)。血气检测结果显示,与空白组相比,模型组pH显著降低(P<0.05),且二氧化碳分压(PCO_(2))、碳酸氢根(HCO_(3)^(-))、阴离子间隙(AnGap)、二氧化碳总量(tCO_(2))和钾离子(K^(+))含量均显著升高(P<0.05),模型组和各给药组的氯离子(Cl^(-))和钠离子(Na^(+))含量显著升高(P<0.05)。抗氧化指标检测结果显示,与空白组相比,模型组超氧化物歧化酶(SOD)活性和葡萄糖(GLU)含量显著降低(P<0.05),乳酸(LD)含量显著升高(P<0.05),各给药组的SOD活性均显著降低(P<0.05),低剂量组和中剂量组GLU含量显著降低(P<0.05),各给药组的丙二醛(MDA)和LD含量差异不显著(P>0.05)。结果表明,服用生脉饮加味可改善运动性疲劳大鼠的运动性能、延长运动时间,可降低心肌酶谱中酶的活性,改善酸碱失衡和氧化应激造成的运动性疲劳,进而发挥抗疲劳和保护心脏功能。本试验结果为缓解运动疲劳相关药物的研制提供了参考依据。

地榆皂苷Ⅰ对大鼠膝骨关节炎的作用机制研究

目的研究地榆皂苷Ⅰ对大鼠膝骨关节炎的作用机制。方法选择SD雄性乳鼠8只,提取乳鼠软骨细胞,采用CCK-8实验检测细胞增殖率以确定最佳药物浓度。将软骨细胞随机分为对照组、炎症组和治疗组,对照组在完全培养基中进行培养,炎症组在含有10μg/mL白细胞介素-1β(IL-1β)的培养基中培养,治疗组在含有10μg/mL IL-1β和最佳浓度地榆皂苷Ⅰ的培养基中培养。采用Calcein-AM/PI染色观察各组软骨细胞生长情况。采用实时荧光定量聚合酶链式反应(RT-qPCR)检测各组软骨细胞中白细胞介素-6(IL-6)、基质金属蛋白酶13(MMP13)mRNA相对表达量。取8周龄左右的SD雄性大鼠24只随机分为对照组、炎症组和治疗组,每组8只,对炎症组和治疗组建立骨关节炎模型,建模完成4周后,炎症组和对照组的关节腔内注射等体积生理盐水,治疗组的关节腔内注射100μL最佳浓度的地榆皂苷Ⅰ溶液,1次/周,连续给药4周。采用HE染色和番红O-固绿染色观察各组膝关节组织结构。采用免疫组化染色观察各组膝关节组织中IL-6、MMP13蛋白表达情况。结果CCK-8实验结果显示,各实验浓度均未表现明显细胞毒性,当地榆皂苷Ⅰ的浓度为20 mg/L时,软骨细胞增殖率最高。Calcein-AM/PI染色结果显示,炎症组和治疗组细胞增殖率低于对照组,治疗组细胞增殖率高于炎症组,差异有统计学意义(P<0.05)。RT-qPCR检测结果显示,炎症组和治疗组IL-6、MMP13 mRNA相对表达量高于对照组,治疗组IL-6、MMP13 mRNA相对表达量低于炎症组,差异有统计学意义(P<0.05)。HE染色和番红O-固绿染色结果显示,对照组的膝关节组织结构比炎症组和治疗组完整,与炎症组相比,治疗组膝关节组织损伤有所减轻。免疫组化染色结果显示,炎症组IL-6、MMP13蛋白表达水平最高,治疗组IL-6、MMP13蛋白表达水平高于对照组。结论地榆皂苷Ⅰ可通过维持软骨细胞活力、降低炎症因子的释放以及减少软骨细胞外基质(ECM)的降解发挥抗炎作用,对膝骨关节炎有一定的药用潜力。

炒白芍-炙甘草配伍前后化学成分与抗炎作用相关性研究

目的明确炒白芍-炙甘草配伍对化学成分的影响及其与抗炎作用的相关性。方法采用HPLCDAD建立同时测定炒白芍-炙甘草中芍药苷、芍药内酯苷、苯甲酰芍药苷、氧化芍药苷、甘草酸、甘草苷多成分含量的分析方法;建立佐剂性关节炎大鼠模型,给药组分别灌胃炒白芍、炙甘草、炒白芍-炙甘草药液2周,ELISA检测大鼠血清炎症因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、前列腺素E_(2)(PGE_(2))含量;采用析因设计、双变量相关分析、多元线性回归方法研究配伍对化学成分的影响及其与抗炎药效的相关性。结果建立HPLC-DAD分析方法,炒白芍-炙甘草配伍后氧化芍药苷、芍药内酯苷、苯甲酰芍药苷含量增加(P<0.05);与模型组比较,各给药组大鼠血清IL-1β、TNF-α、PGE_(2)含量降低(P<0.01),且配伍组血清IL-1β、TNF-α、PGE_(2)含量低于炒白芍组、炙甘草组(P<0.05,P<0.01);析因设计结果显示炒白芍-炙甘草配伍存在交互效应,双变量相关分析结果显示氧化芍药苷、芍药内酯苷、芍药苷、甘草苷、苯甲酰芍药苷、甘草酸含量分别与IL-1β、TNF-α、PGE_(2)含量呈负相关,多元线性回归结果显示甘草苷、甘草酸与IL-1β、TNF-α、PGE_(2)含量呈负相关。结论本研究建立的炒白芍-炙甘草多成分含量分析方法简便快捷、专属性强、准确可靠,可用于其质量评价与控制。炒白芍-炙甘草配伍可抑制炎症因子IL-1β、TNF-α、PGE_(2)表达,且化学成分含量与抗炎作用具有相关性。

Anti-TNF-αMcAb对创伤失血性休克大鼠肾脏保护作用的实验研究

目的 :研究肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和抗肿瘤坏死因子-α单克隆抗体(anti-TNF-αmonoclonal antibody,anti-TNF-αMc Ab)在创伤失血性休克大鼠肾脏损害中的作用。方法:24只成年雄性SD大鼠随机均分为Ⅰ组(对照组)、Ⅱ组(休克用乳酸林格氏液治疗组)和Ⅲ组(休克用anti-TNF-αMc Ab治疗组)。Ⅱ组、Ⅲ组用电刀在大鼠腹正中切开约5 cm长的切口,股动脉放血,制作创伤失血性休克动物模型;Ⅱ组用乳酸林格氏液治疗,Ⅲ组用含anti-TNF-αMc Ab(3 mg/kg)的乳酸林格氏液治疗,而Ⅰ组在相同条件下不进行创伤和失血。测定所有SD大鼠血清肌酐(creatinine,Cr)、尿素氮(urea nitrogen,UN)、TNF-α、肾组织中丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)水平。在光镜、电镜下观察所有大鼠肾组织的病理学改变情况。结果 :Ⅱ、Ⅲ组大鼠血清Cr、UN、TNF-α、肾组织中MDA含量较Ⅰ组升高(P<0.05),SOD含量减低(P<0.05);Ⅲ组Cr(133.10±17.39)μmol/L、UN(23.78±5.49)mmol/L、TNF-α(106.85±14.65)pg/m L、肾组织中MDA(13.26±1.46)nmol/mgprot水平比Ⅱ组血清Cr、UN、TNF-α、肾组织中MDA水平降低(P<0.05),Ⅲ组SOD(79.26±11.21)U/mgprot水平比Ⅱ组增高(P<0.05)。Ⅲ组大鼠肾组织病理损伤比Ⅱ组轻。Ⅰ组无明显病理改变。结论:TNF-α可能是创伤失血性休克大鼠肾损害的重要介导因子,使用anti-TNF-αMc Ab治疗可以减轻创伤失血性休克时大鼠肾损害,为临床治疗提供新的策略。

灯盏细辛总咖啡酰奎宁酸调控PI3K/Akt/FoxO3a通路延缓衰老的作用机制研究

研究灯盏细辛总咖啡酰奎宁酸(caffeoylquinic acid from Erigeron breviscapus,EBCQA)的抗衰老作用,并探讨其潜在作用机制。以线虫为模型,观察EBCQA对其寿命、氧化抗性、热抗性、运动能力、DAF-16入核、以及超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)活性和丙二醛(malondialdehyde,MDA)含量的影响。腹腔注射D-半乳糖构建衰老大鼠模型,检测EBCQA对大鼠学习记忆能力、胸腺和脾脏系数、肝脏中磷脂酰肌醇3-激酶(phosphoinositide 3-kinase,PI3K)、蛋白激酶B(Akt/protein kinase B,Akt)、磷酸化Akt(phosphp-Akt,p-Akt)、叉头框蛋白O3a(forkhead box class O3a,FOXO3a)、磷酸化FOXO3a(phosphp-FOXO3a,p-FOXO3a)表达,以及肝脏和血清中SOD、GSH-Px、CAT活性和MDA含量的影响。结果显示,EBCQA能够延长野生型线虫寿命,并提升其氧化抗性、热抗性和运动能力,但对线虫PI3K、Akt、FoxO3a同源体突变株寿命没有显著影响;EBCQA能促进DAF-16入核,通过DAF-16提升线虫的SOD、GSH-Px、CAT活性,降低MDA水平。在D-半乳糖诱导衰老大鼠中,EBCQA给药显著改善大鼠的学习记忆功能,提升胸腺和脾脏系数,降低PI3K、Akt、p-Akt、p-FoxO3a表达和MDA含量,增强SOD、GSH-Px、CAT活性。以上结果表明,EBCQA具有延缓衰老的作用,其机制可能与抑制PI3K和Akt激活、降低FOXO3a磷酸化、促进FOXO3a入核和转录活性、提升抗氧化酶活性和抑制氧化应激相关。

巴马火麻油对大鼠抗氧化功能的影响

为研究巴马火麻油对大鼠抗氧化功能的影响,依据《关于印发抗氧化功能评价方法等9个保健功能评价方法的通知》(国食药监保化[2012]107号)附件1抗氧化功能评价方法,采用巴马火麻油对成年大鼠进行实验,设溶媒对照组、空白对照组及火麻油高、中、低3个剂量组,连续灌胃30 d。结果表明,受试样品组肝组织匀浆和血清中的丙二醛低于溶媒对照组,差异显著(P<0.05),肝组织匀浆中的超氧化物歧化酶活性极显著高于溶媒对照组(P<0.01);受试样品组肝组织匀浆和血清中的还原型谷胱甘肽含量高于溶媒对照组,差异极显著(P<0.01)。这表明在本实验条件下,动物实验的脂质过氧化产物、抗氧化酶、抗氧化物质3项指标阳性,表明火麻油具有抗氧化功能。