Hepatoprotective effect of manual acupuncture at acupoint GB34 against CCl_4-induced chronic liver damage in rats

AIM： To investigate the hepatoprotective effect of manual acupuncture at Yanglingquan （GB34） on CCl4-induced chronic liver damage in rats. METHODS： Rats were injected intraperitoneally with CCh （1 mL/kg） and treated with manual acupuncture using reinforcing manipulation techniques at left GB34 （Yanglingquan） 3 times a week for 10 wk. A nonacupoint in left gluteal area was selected as a sham point. To estimate the hepatoprotective effect of manual acupuncture at GB34, measurement of liver index, biochemical assays including serum ALT, AST, ALP and total cholesterol, histological analysis and blood cell counts were conducted. RESULTS： Manual acupuncture at GB34 reduced the liver index, serum ALT, AST, ALP and total cholesterol levels as compared with the control group and the sham acupuncture group. It also increased and normalized the populations of WBC and lymphocytes. CONCLUSION： Manual acupuncture with reinforcing manipulation techniques at left GB34 reduces liver toxicity, protects liver function and liver tissue, and normalizes immune activity in CCh-intoxicated rats.

Effect of hepatocyte growth factor on inflammatory factors associated with CCL_(4)-induced hepatocyte injury

Objective:The aim of this study is to investigate the effects of Hepatocyte Growth Factor(HGF)on the expression levels of IL-8,TNF-α,IL-4,and IL-21 in mice with liver injury induced by CCL_(4).Methods:An acute liver injury mouse model was established using CCL_(4),and hepatocytes and white blood cells were separated by gradient density centrifugation.Different concentrations of HGF were added in vitro,and the expression levels of cytokines were detected using ELISA.Results:In the in vivo injury model,the hepatocyte experiment results showed that the expression level of IL-8 was reduced in the 10 ng/mL HGF group compared to the injured hepatocyte group(P<0.05),and increased in the 50 ng/mL HGF group compared to the 10 ng/mL HGF group(P<0.05).For IL-4,the expression levels were reduced in both the 25 ng/mL HGF group(P<0.05)and the 50 ng/mL HGF group(P<0.05)compared to the injured hepatocyte group.The white blood cell experiment results showed that the expression levels of TNF-αwere reduced in both the 10ng/ml HGF group(P<0.05)and the 25 ng/mL HGF group(P<0.05)compared to the injured white blood cell group.In the in vitro injury model,hepatocyte experiment results showed that the expression levels of TNF-αwere reduced in both the 25 ng/mL HGF group(P<0.05)and the 50 ng/mL HGF group(P<0.05)compared to the normal control group.For IL-4,the expression level was reduced in the 25 ng/mL HGF group compared to the normal control group(P<0.05).The white blood cell experiment results showed that the expression level of TNF-αwas increased in the 50 ng/mL HGF group compared to the 10 ng/mL HGF group(P<0.001);for IL-21,the expression levels were reduced in the CCL_(4) model group(P<0.05),10 ng/mL HGF group(P<0.05),25 ng/mL HGF group(P<0.05),and 50 ng/mL HGF group(P<0.05)compared to the normal control group.Conclusion:when the liver of mice is acutely damaged by CCL_(4),HGF can reduce the expression levels of inflammatory cytokines IL-8,TNF-α,IL-4 in hepatocytes,and TNF-αin liver white blood cells.

Protective Effects of Flavonoids from Pteridium aquilinum on CCl_(4)-induced Acute Liver Injury in Mice

[Objectives]To explore the protective effects of flavonoids from Pteridium aquilinum(PAFL)on carbon tetracholoride(CCl_(4))-induced acute liver injury in mice and its potential mechanism.[Methods]All mice were randomly divided into four groups(n=10 in each),normal group,CCl_(4)group,CCl_(4)+PAFL groups[treated with PAFL(50 or 200 mg/kg)].Animal treatment was continued for 7 consecutive days.The blood was collected after injection of CCl_(4)for 24 h,and the liver tissue was removed from the mice and stored at-80℃.[Results]The PAFL(50 and 200 mg/kg)significantly inhibited the increase of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)levels in serum caused by CCl_(4)treatment.PAFL administration not only increased the activity of antioxidant enzymes superoxide dismutase(SOD),Glutathione(GSH)and catalase(CAT)in mice,but also reduced the level of malondialdehyde(MDA).Meanwhile,PAFL administration decreased the expression of nuclear factor-kappa B(NF-κB)and Cyclooxygenase-2(COX-2)proteins and inhibited the release of pro-inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin 6(IL-6).In addition,PAFL(200 mg/kg)treatment down-regulated extracellular regulated protein kinases(ERK)and c-Jun N-terminal kinase(JNK)protein levels in liver tissue.[Conclusions]These findings clearly indicate that the protective effects of PAFL on CCl_(4)-induced acute liver injury is related to its antioxidant and anti-inflammatory activity,which may be mediated by NF-κB and MAPKs signaling pathways.

合生元对CCl_(4)诱发小鼠急性化学性肝损伤的改善作用及其机制

目的探讨不同组合的合生元对CCl_(4)所致急性化学性肝损伤小鼠的肝脏保护作用,及其机制是否与肠—肝轴有关。方法将72只SPF级ICR雄性小鼠随机分12组,每组6只。将含量为5%、15%和20%的麦麸膳食纤维饲料分别记为A1、A2、A3,灌胃剂量为10、20、30 mL/kg的益生菌酸奶分别记为B1、B2、B3,对A1B1组、A1B2组、A1B3组、A2B1组、A2B2组、A2B3组、A3B1组、A3B2组、A3B3组分别进行不同组成合生元干预,阳性对照组、模型组和空白对照组给予普通饲料,阳性对照组同时灌胃复方鳖甲软肝片溶液;30 d后,除空白对照组外的其余各组小鼠灌胃给予1%的CCl_(4)油溶液5 mL/kg,空白对照组灌胃给予等体积玉米油溶液。24 h后,全部小鼠摘眼球取血并分离血清,采用ELISA法检测血清超氧化物歧化酶(SOD)、丙二醛(MDA)水平;取肝脏、结肠、回肠,计算肝脏指数和结肠指数,同时对肝脏和回肠进行HE染色后观察病理改变;收集A1B1组、阳性对照组、模型组和空白对照组小鼠粪便,采用16S rDNA高通量测序法检测肠道菌群情况(Chao、Ace、Shannon、Simpson指数及门水平、属水平上的肠道菌群丰度)。结果模型组肝脏指数及血清MDA水平均高于空白对照组(P均<0.05)。A1B1、A1B2、A2B1和A3B1组小鼠肝脏指数均低于模型组,A1B1、A2B1组小鼠肝脏指数均低于阳性对照组(P均<0.05)。各组小鼠结肠指数比较差异无统计学意义(P>0.05)。A2B1、A2B3组小鼠血清SOD水平均低于模型组、阳性对照组,A1B1、A1B2、A2B3、A3B1和A3B2组小鼠血清MDA水平均低于模型组(P均<0.05);除A1B2组外,其余合生元组小鼠血清MDA水平均高于阳性对照组(P均<0.05)。肝脏病理观察结果:空白对照组小鼠肝组织细胞排列紧密且形态正常,细胞结构清晰且完整,未见炎症细胞浸润和坏死细胞;模型组小鼠肝索、肝小叶结构严重紊乱,细胞肿胀、细胞内存在大量未代谢的CCl_(4);阳性对照组肝细胞形态正常,仅有少量未代谢的CCl_(4);与模型组比较,A1B1组肝小叶结构完整、仅有少量炎症细胞浸润,A2B2组、A3B3组肝小叶严重受损、有大量炎症细胞浸润,A3B1组和A3B3组可见大量未代谢的CCl_(4)。回肠病理观察结果:空白对照组小鼠回肠绒毛较长,绒毛结构完整,肠内皮结构较厚;模型组小鼠回肠结构破坏,肠腔内可见大量黑色物质;阳性对照组绒毛结构较为完整,肠腔内有少量黑色物质;与空白对照组相比,其他合生元组均存在绒毛结构破坏,以A1B1组小鼠绒毛结构完整度最高。模型组小鼠Chao、Ace、Shannon指数均低于空白对照组及A1B1组,Simpson指数高于空白对照组及A1B1组;阳性对照组Shannon指数高于模型组,Simpson指数低于模型组(P均<0.05)。门水平上:与空白对照组比较,模型组小鼠拟杆菌门和变形菌门相对丰度升高,而厚壁菌门相对丰度降低;与模型组比较,A1B1组小鼠厚壁菌门和变形菌门相对丰度均升高,拟杆菌门相对丰度均降低;与阳性对照组比较,A1B1组疣微菌门相对丰度降低(P均<0.05)。属水平上:模型组有10种菌属的相对丰度低于空白对照组,A1B1组7种菌属的相对丰度高于模型组(P均<0.05)。结论合生元对CCl_(4)所致急性化学性肝损伤小鼠具有一定的肝脏保护作用,以5%麦麸膳食纤维联合10 mL/kg益生菌酸奶的合生元组合的肝脏保护效果更好,其机制可能与调节肠—肝轴有关。

肝细胞生长因子对CCl_(4)致肝细胞损伤相关炎性因子的影响

目的:探究肝细胞生长因子(hepatocyte growth factor,HGF)对CCl_(4)所致小鼠肝脏损伤IL⁃8、TNF⁃α、IL⁃4、IL⁃21表达水平的影响。方法:建立CCl_(4)急性肝损伤小鼠模型,通过梯度密度离心分离肝实质细胞及白细胞,体外加入不同浓度HGF,ELISA检测细胞因子表达水平。结果:体内损伤模型中,肝实质细胞实验结果显示,10 ng/mL HGF组与损伤肝细胞组相比IL⁃8表达水平降低(P<0.05),50 ng/mL HGF组与10 ng/mL HGF组相比IL⁃8表达水平升高(P<0.05);与损伤肝细胞组相比,25 ng/mL HGF组(P<0.05),50 ng/mL HGF组(P<0.05)IL⁃4表达水平均降低。白细胞实验结果显示,与损伤白细胞组相比,10 ng/mL HGF组(P<0.05)、25 ng/mL HGF组(P<0.05)TNF⁃α表达水平均降低。体外损伤模型中,肝实质细胞实验结果显示,与正常对照组相比,25 ng/mL HGF组(P<0.05)、50 ng/mL HGF组(P<0.05)TNF⁃α表达水平均降低;25 ng/mL HGF组与正常对照组相比IL⁃4表达水平降低(P<0.05)。白细胞实验结果显示,50 ng/mL HGF组与10 ng/mL HGF组相比TNF⁃α表达水平升高(P<0.001);与正常对照组相比CCl_(4)模型组(P<0.05)、10 ng/mL HGF组(P<0.05)、25 ng/mL HGF组(P<0.05)、50 ng/mL HGF组(P<0.05)IL⁃21表达水平均降低。结论:CCl_(4)致小鼠肝脏急性受损时,HGF可降低肝实质细胞分泌炎性因子IL⁃8,TNF⁃α,IL⁃4以及肝脏白细胞TNF⁃α的表达水平。

基于TGF-β1/Smads信号通路探讨2-十二烷基-6-甲氧基-2,5-二烯-1,4-环己二酮抗肝纤维化的作用机制

目的探讨2-十二烷基-6-甲氧基-2,5-二烯-1,4-环己二酮(2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione,DMDD)对CCl_(4)致大鼠肝纤维化的影响及潜在作用机制。方法采用50%CCl_(4)复制肝纤维化模型,检测血液中门冬氨酸转移酶(aspartate transferase,AST)、丙氨酸转移酶(alanine transferase,ALT)、白蛋白/球蛋白(albumin/globulin,A/G)、总蛋白(total protein,TP)、总胆红素(total bilirubin,T-BIL)、大鼠透明质酸(hyaluronic acid,HA)、层黏连蛋白(laminin,LN)、Ⅲ型胶原(collagen typeⅢ,ColⅢ)和Ⅳ型胶原(collagen typeⅣ,ColⅣ)。HE和Masson染色观察肝脏组织病变和纤维形成情况。免疫组织化学法检测肝脏组织中α平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)、Ⅰ型胶原(collagen typeⅠ,ColI)、转化生长因子β1(transformed growth factor-β1,TGF-β1)、Smad2、Smad7蛋白表达水平。RT-PCR法检测肝脏组织中α-SMA、ColI、TGF-β1、Smad7mRNA水平。Western blot法检测肝脏组织中TGF-β1、Smad4、Smad7蛋白表达水平。结果与正常对照组相比,模型组血清AST、ALT、TP、T-BIL、HA、LN、ColⅢ、ColⅣ水平明显升高,A/G水平明显降低,肝脏HE和Masson染色显示有大量纤维形成,说明造模成功。与模型组相比,DMDD给药组大鼠AST、ALT、TP、T-BIL、HA、LN、ColⅢ、ColⅣ含量和α-SMA、ColI、TGF-β1、Smad2、Smad4mRNA及蛋白水平显著下调,A/G、Smad7水平上调。肝脏HE和Masson染色结果显示纤维增生明显减少。结论DMDD对CCl_(4)诱导的肝纤维化具有保护作用,其机制可能与TGF-β1/Smads信号通路有关。

电针联合乌司他丁对CCl_(4)诱导大鼠急性肝损伤的保护作用及对TLR4/NF-κB信号通路的影响

目的探讨电针联合乌司他丁对四氯化碳(CCl_(4))诱导大鼠急性肝损伤的保护作用及对Toll样受体(TLR)4/核因子(NF)-κB信号通路的影响。方法60只大鼠中随机选取12只作为正常组,采用CCl_(4)处理建立急性肝损伤模型,选取造模成功的大鼠随机分为模型组、电针组、乌司他丁组、联合组,每组各12只。检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、白细胞介素(IL)-6、IL-12、肿瘤坏死因子(TNF)-α水平,计算肝指数;检测肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)水平;采用酶联免疫吸附试验(ELISA)检测IL-6、IL-12、TNF-α水平;采用Western印迹检测B细胞淋巴瘤-2相关蛋白(Bax)、B细胞淋巴瘤(Bcl)-2、TLR4、p-p65、p65蛋白表达。结果与正常组比较,模型组肝指数显著降低,血清ALT、AST、IL-6、IL-12、TNF-α、MDA水平、Bax、TLR4、p-p65显著升高,SOD水平、Bcl-2显著降低(均P<0.05);与模型组比较,电针组、乌司他丁组、联合组大鼠肝指数显著升高,血清ALT、AST、IL-6、IL-12、TNF-α、MDA水平、Bax、TLR4、p-p65显著降低,SOD水平、Bcl-2显著升高(均P<0.05),且联合组作用效果显著优于电针组、乌司他丁组(P<0.05)。结论电针联合乌司他丁联合治疗可降低炎症反应、氧化应激及抑制凋亡从而对CCl_(4)诱导大鼠急性肝损伤发挥保护作用,其机制可能与TLR4/NF-κB通路被抑制有关。

Antioxidant activity and hepatoprotective effect of 10 medicinal herbs on CCl4-induced liver injury in mice

BACKGROUND Many natural products confer health benefits against diverse diseases through their antioxidant activities.Carbon tetrachloride(CCl4)is often used in animal experiments to study the effects of substances on liver injury and the related mechanisms of action,among which oxidative stress is a major pathogenic factor.AIM To compare antioxidant and hepatoprotective activities of ten herbs and identify and quantify phytochemicals for the one with strongest hepatoprotection.METHODS The antioxidant activity of ten medicinal herbs was determined by both ferricreducing antioxidant power and Trolox equivalent antioxidant capacity assays.The total phenolic and flavonoid contents were determined by Folin–Ciocalteu method and aluminum chloride colorimetry,respectively.Their effects on CCl4-induced oxidative liver injury were evaluated and compared in a mouse model by administrating each water extract(0.15 g/mL,10 mL/kg)once per day for seven consecutive days and a dose of CCl4 solution in olive oil(8%,v/v,10 mL/kg).The herb with the strongest hepatoprotective performance was analyzed for the detailed bioactive components by using high-performance liquid chromatography-electrospray ionization source-ion trap tandem mass spectrometry.RESULTS The results revealed that all tested herbs attenuated CCl4-induced oxidative liver injury;each resulted in significant decreases in levels of serum alanine transaminase,aspartate transaminase,alkaline phosphatase,and triacylglycerols.In addition,most herbs restored hepatic superoxide dismutase and catalase activities,glutathione levels,and reduced malondialdehyde levels.Sanguisorba officinalis(S.officinalis)L.,Coptis chinensis Franch.,and Pueraria lobata(Willd.)Ohwi root were the three most effective herbs,and S.officinalis L.exhibited the strongest hepatoprotective effect.Nine active components were identified in S.officinalis L.Gallic acid and(+)-catechin were quantified(7.86±0.45 mg/g and 8.19±0.57 mg/g dried weight,respectively).Furthermore,the tested herbs displayed a range of in vitro antioxidant activities proportional to their phenolic content;the strongest activities were also found for S.officinalis L.CONCLUSION This study is of value to assist the selection of more effective natural products for direct consumption and the development of nutraceuticals or therapeutics to manage oxidative stress-related diseases.

The new antioxidant 1-benzoyl-6-hydroxy-2,2,4-trimethyl-1,2- dihydroquinoline has a protective effect against carbon tetrachloride-induced hepatic injury in rats

Liver diseases with the central pathogenetic mechanism of oxidative stress are one of the main causes of mortality worldwide.Therefore,dihydroquinoline derivatives,which are precursors of hepatoprotectors and have antioxidant activity,are of interest.We have previously found that some compounds in this class have the ability to normalize redox homeostasis under experimental conditions.Here,we initially analyzed the hepatoprotective potential of the dihydroquinoline derivative 1-benzoyl-6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline(BHDQ)for carbon tetrachloride(CCl4)-induced liver injury in rats.Results suggested that BHDQ normalized the alanine aminotransferase,aspartate aminotransferase,and gamma-glutamyl transpeptidase in serum.We also observed an improvement in liver tissue morphology related to BHDQ.Animals with CCl4-induced liver injuries treated with BHDQ had less oxidative stress compared to animals with CCl4-induced liver injury.BHDQ promoted activation changes in superoxide dismutase,catalase,glutathione peroxidase,glutathione reductase,and glutathione transferase on control values in animals with CCl4-induced liver injury.BHDQ also activated gene transcription in Sod1 and Gpx1 via nuclear factor erythroid 2-related factor 2 and forkhead box protein O1 factors.Therefore,the compound of concern has a hepatoprotective effect by inhibiting the development of necrotic processes in the liver tissue,through antioxidation.

应用Micro CT观察洛伐他汀和秋水仙碱对CCl_4致肝损伤后小鼠骨代谢的影响

目的本实验应用Micro CT研究洛伐他汀和秋水仙碱对CCl4致肝损伤后小鼠骨代谢的影响,并比较二者的效果。方法体积分数40%的CCl4花生油皮下注射,建立小鼠肝损伤模型,同时,洛伐他汀按2.6 mg·kg-1,秋水仙碱按0.065 mg·kg-1每天1次灌胃给药,连续1个月,于实验结束后观察肝损伤血清学指标以及肝匀浆相关的抗氧化指标,并用Micro CT测定小鼠胫骨骨组织结构参数。结果单用CCl4小鼠肝指数明显增加,AST和ALT活性明显升高,肝匀浆中SOD活性和GSH-Px水平明显降低,MDA水平明显升高,且胫骨上段骨组织结构参数BVF、TMD值明显降低,SMI值明显升高,出现明显的骨质疏松症状。而洛伐他汀、秋水仙碱用药后对肝损伤没有明显影响,但洛伐他汀对肝损伤导致的骨丢失有明显的保护作用,秋水仙碱组则没有明显的保护作用。结论 CCl4致肝损伤后的小鼠出现骨质疏松症状,洛伐他汀用药后对CCl4致肝损伤的骨丢失有明显的预防作用。

鲨肝刺激物质类似物对CCl_4致小鼠急性肝损伤的保护作用

目的:探讨重组鲨肝刺激物质类似物(r-sHSA)对CCl4致小鼠急性肝损伤的保护作用及其作用机制.方法:利用腹腔注射CCl4制备小鼠急性肝损伤动物模型,以血清转氨酶活性以及组织病理变化为指标判断r-sHSA对化学性肝损伤的保护作用,并通过定量RT-PCR方法测定肝组织中相关细胞因子表达量的变化.结果:8～200μg/kg r-sHSA均可显著降低损伤小鼠血清转氨酶的活性,明显减轻肝细胞肿胀,减少中性粒细胞浸润,具有显著的肝保护作用,其机制可能与r-sHSA诱导TNF-a和肝细胞生长因子(HGF)表达,并下调TGF-β1的表达有关.结论:r-sHSA对CCl4致小鼠急性肝损伤具有明显的保护作用,其机制可能与细胞因子的表达调控有关.

人羊膜间充质干细胞移植对CCl_4诱导的小鼠损伤肝HGF、SIRT-1、α-SMA及P^(27kip1)表达的影响

人羊膜间充质干细胞(h AMSCs)具有自我增殖和多向分化潜能,有望为干细胞移植性治疗提供新来源,是病变组织器官损伤修复的理想种子细胞.但目前关于h AMSCs对肝损伤的修复机制仍不十分清楚.本研究采用胰蛋白酶-胶原酶消化法从羊膜组织中分离、纯化了间充质干细胞.免疫荧光检测表面标记波形丝蛋白(vimentin)和阶段特异表达抗原4(SSEA-4)均呈阳性.h AMSCs表达CD29、CD49d、CD73表面抗原,但不表达骨髓间充质表面抗原CD34、CD45和人类白细胞抗原DR位点(HLA-DR).实时定量PCR和Western印迹检测揭示,h AMSCs移植后可提高受损肝组织中肝细胞生长因子(HGF)和沉默信息调节因子1(SIRT-1)的表达,抑制α-平滑肌肌动蛋白(α-SMA)和周期性蛋白依赖性激酶抑制因子(P27kip1)的表达.因为上述蛋白质分子涉及肝细胞增殖、再生、凋亡调节,抑或肝纤维化过程,因此h AMSCs移植后所引起的上述分子表达变化可改善四氯化碳(CCL4)诱导的肝损伤,抑制肝细胞凋亡,促进肝细胞有丝分裂,对肝损伤有一定的修复作用.该研究为进一步探索调控肝再生、损伤修复信号通路(机制)及预防肝纤维化提供了新启示.

CCl_(4)和HCl气体氯化铀氧化物

分别以CCl_(4)和HCl气体作为氯化试剂,进行了铀氧化物(主要为U_(3)O_(8))的氯化机理和各影响因素研究。以CCl_(4)为氯化试剂对U_(3)O_(8)粉末进行氯化,通过热重分析研究了氯化反应过程的机理及动力学行为,氯化产物主要为UCl_(4)。同时研究了CCl_(4)对不同种类和形态铀氧化物的氯化,UO_(2)芯块由于结构致密很难进行氯化,UO_(2)粉末和UO_(3)粉末很容易被CCl_(4)氯化,产物分别为UCl_(4)和UCl_(6)。以HCl气体为氯化试剂对LiCl-KCl熔盐中的U_(3)O_(8)粉末进行氯化,研究了反应温度、氯化时间、HCl气体流速、U_(3)O_(8)粉末投料量以及铀氧化物种类和形态的影响。结果表明,提高反应温度、延长反应时间、提高HCl气体流速,有利于氯化率的提高。推荐HCl气体氯化U_(3)O_(8)粉末的工艺参数为:氯化反应温度为500℃、HCl气体流速为0.6 L/min。

Hepatoprotective effect of Premna corymbosa(Burm.f.) Rottl.& Willd.leaves extract on CCl_4 induced hepatic damage in Wistar albino rats

Objective:To investigate the hepetoprotective activity of Premna corymbosa leaves against carbon tetrachloride(CCl_4) induced hepatic damage.Methods:Hepatotoxicity was induced in wistar rats of both sexes by intraperitoneal injection of CCl_4,1 mL/kg body weight for every 72 h.The ethanolic extract of Premna corymbosa leaves were administrated at doses of 200 & 400 mL/kg body weight, p.o.,daily for 14 days.The hepatotoxicity and its prevention was assessed by serum markers like serum alkaline phosphatase(SALP),serum triglycerides(STG),serum total protein(STP), serum cholesterol(SC),and liver wet weight and histopathological studies of the liver.Results:In treatment with the ethanolic extract,the toxic effect of CCl_4 was controlled significantly(P<0.01) by restoration of the levels of biochemical parameters as compared to normal and standard drug silymarin treated groups.The liver weight was reduced by the ethanolic extract treated groups. The histopathology of the liver sections evidenced the hepatoprotective activity.Conclusion:The ethanolic extract of the leaves of Premna corymbosa possess significant acute hepatoprotective activity.Premna corymbosa can be recommended for the liver disorders.

Hedgehog通路介导上皮-间质转化促进CCl_(4)诱导的大鼠肝纤维化进展

[目的]观察四氯化碳(carbon tetrachloride,CCl4)诱导大鼠肝纤维化发生发展过程中,Hedgehog信号通路及上皮-间质转化(epithelial-mesenchymal transition,EMT)关键分子的变化情况,探讨其可能的作用机制。[方法]将64只雄性无特殊病原体(specific pathogen free,SPF)级SD大鼠随机分为正常组和模型组,每组32只。模型组大鼠皮下注射橄榄油稀释的40%CCl43mL·kg-1,正常组用等容量的橄榄油代替CCl4,2次/周,首次加倍,共6周,建立大鼠肝纤维化模型,于2、7、21、42d采用全自动生化仪检测血清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)、总胆红素(total bilirubin,TBIL)水平,以碱水法检测肝组织羟脯氨酸(hydroxyproline,Hyp)含量,苏木精-伊红(hematoxylin-eosin,HE)染色和Masson染色观察肝组织炎症及纤维化程度,实时荧光定量聚合酶链式反应(quantitative Real-time polymerase chain reaction,qRT-PCR)和Western blot检测肝组织Hedgehog信号通路及EMT关键分子上皮-钙粘素(epithelial-cadherin,E-cadherin)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、平滑受体(smoothened,Smo)及神经胶质瘤相关癌基因同源蛋白-1(glioma-associated oncogene homolog-1,Gli-1)mRNA和蛋白表达。[结果]模型组大鼠血清ALT、AST、ALP、TBIL水平和肝组织Hyp含量随时间延长逐渐增高,21、42d均显著高于正常组同期(P<0.01)。肝组织炎症和纤维化程度也随时间延长而逐渐加重,均较正常组同期显著增高(P<0.05,P<0.01)。模型组肝脏E-cadherin mRNA和蛋白表达随时间延长而逐渐下降,α-SMA、Smo、Gli-1的mRNA和蛋白表达则逐渐上升(P<0.05,P<0.01),且21、42d与正常组同期差异均有统计学意义(P<0.05,P<0.01)。[结论]CCl4诱导的大鼠肝纤维化发生过程中存在EMT和Hedgehog信号通路异常激活,Hedgehog信号通路可能通过介导EMT,从而促进肝纤维化进展。

双金花茶对CCl_(4)致小鼠急性肝损伤的保护作用

为了研究双金花茶对CCl_(4)致小鼠急性肝损伤的保护作用,将50只KM小鼠随机分为5组,双金花茶低、高剂量组(2.59、10.35 g/kg)每天灌胃1次,连续14 d,空白对照组、溶媒对照组和模型组给予等体积蒸馏水灌胃。末次给药后,模型组和双金花茶组小鼠给予25%CCl_(4)皮下注射,24 h后处死小鼠,检测小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的活性,肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)的含量;计算肝、脾指数;HE染色观察肝组织病理学变化。与模型组比较,双金花茶高剂量组血清ALT和AST活性显著降低(P<0.01);肝组织MDA含量显著降低(P<0.01),SOD和GSH-Px活性显著升高(P<0.05,P<0.01);肝、脾指数明显降低(P<0.05,P<0.01);肝组织病理性损伤明显减轻。双金花茶对CCl_(4)致小鼠急性肝损伤有一定的保护作用,其机制可能为抑制细胞膜脂质过氧化和清除活性氧自由基,从而增强其抗氧化的作用。

蒲公英总黄酮对CCl_(4)致大鼠肝损伤的保护作用

目的研究蒲公英总黄酮对CCl_(4)致大鼠肝损伤的保护作用。方法144只大鼠随机分为空白组,模型组,联苯双酯组(3.75 mg/kg),蒲公英总黄酮低、中、高剂量组(50、100、200 mg/kg),每组24只,分别灌胃给予相应药物(10 mL/kg),连续7 d。末次给药后,除空白组外,其余各组大鼠腹腔注射10%CCl_(4)橄榄油溶液10 mL/kg进行造模,于3、6、12、24 h各取6只,检测血清ALT、AST、ALP、TBIL、Arg-1、α-GST、PNP、GLDH水平,以及肝组织匀浆中GSH、MDA水平及SOD活性,RT-qPCR法检测血清及肝组织中miR-122 mRNA表达,HE染色观察肝组织病理变化。结果与空白组比较,模型组大鼠血清ALT、AST、ALP、TBIL、Arg-1、α-GST、PNP、GLDH水平及肝组织MDA水平升高(P<0.05,P<0.01),血清及肝组织miR-122 mRNA表达升高(P<0.05,P<0.01),肝组织SOD活性及GSH水平降低(P<0.05,P<0.01);与模型组比较,蒲公英总黄酮能不同程度地降低大鼠血清ALT、AST、ALP、TBIL、Arg-1、α-GST、PNP、GLDH水平及肝组织MDA水平(P<0.05,P<0.01),下调miR-122 mRNA表达(P<0.05,P<0.01),升高肝组织SOD活性及GSH水平(P<0.05,P<0.01),改善大鼠肝组织病理学损伤。结论蒲公英总黄酮对CCl_(4)致大鼠肝损伤早期具有一定的预防保护作用。

Protective effect of Sida cordata leaf extract against CCl_4 induced acute liver toxicity in rats

Objective:To investigate the hepatoprotective potential of Sida cordata（Malvaceae）（S.cordata） in experimental rats to validate its traditional claim.Methods:Wister albino rats were divided into 6 groups:Croup 1 served as control;CroupⅡserved as hepatotoxic（CCl4 treated） group; CroupⅢ,Ⅳand V served as（100,200 and 400 mg/kg b.w.） S.cordata leaf extract（SCLE） treated groups;CroupⅥserved as positive control（Silymarin） treated group.Liver marker enzymes serum glutamate oxyloacetic transaminase,serum glutamic pyruvic transaminase,pancreatic enzymatic antioxidants superoxide dismutase（SOD）,lipid peroxidation,catalase（CAT）,reduced glutathione（CSH） were measured and compared along with histopathological studies.Results: Obtained results show that the treatment with SCI.E significantly（P【0.05-【0.001） and dosedependently reduced CCl4 induced elevated serum level of hepatic enzymes.Furthermore, SCLE significantly（up to P【0.001） reduced the lipid peroxidation in the liver tissue and restored activities of defence antioxidant enzymes CSH,SOD and CAT towards normal levels,which was confirmed by the histopathological studies.Conclusions:The results of this study strongly indicate the protective effect of SCLE against CCl4 induced acute liver toxicity in rats and thereby scientifically support its traditional use.

肿瘤坏死因子相关蛋白6缺失减缓CCl_(4)诱发的小鼠肝脏纤维化进程

目的探讨补体Clq/肿瘤坏死因子相关蛋白6(CTRP6)在肝病发生发展中的作用及其可能机制。方法四氯化碳灌胃法构建小鼠肝纤维化模型,比较野生型(CTRP6^(+/+))和敲除型(CTRP6^(-/-))小鼠肝损伤后的病理和生化指标差异;RT-qPCR检测纤维化标志基因(Acta2、Col1a1、Mmp2、TGF-β和TNF-α)和细胞增殖标志基因(Cyclin D、Cyclin E和CDK6)的表达水平。结果与CTRP6^(+/+)小鼠相比,CTRP6^(-/-)小鼠肝细胞表现为轻度的坏死、疏松和肿胀等病变,且肝内超氧化物歧化酶(SOD)活力升高(P<0.01)、丙二醛(MDA)含量降低(P<0.01);CTRP6^(-/-)小鼠肝脏纤维化和细胞增殖标志基因表达显著低于CTRP6^(+/+)小鼠。结论CTRP6基因缺失减缓了CCl_(4)诱导的小鼠肝纤维化进程,作用机制可能是敲除CTRP6影响了肝细胞的增殖。

CuCl_(2)-N-甲基咪唑催化合成2,4,4,4-四氯丁腈

以N-甲基咪唑(N-Mim)与CuCl_(2)·2H_(2)O合成了一系列N-甲基咪唑-CuCl_(2)配合物CuL_(n)Cl_(2)[L为N-甲基咪唑(N-Mim),n=1、2、4]。采用XRD、FTIR及TG对其结构进行了表征。比较了CuL_(n)Cl_(2)催化CCl_(4)与丙烯腈(AN)的原子转移自由基加成(ATRA)反应合成2,4,4,4-四氯丁腈(TBN)的催化性能。考察了反应温度、反应时间、催化剂用量及溶剂的协同催化效应对TBN收率的影响.结果表明,CuL_(2)Cl_(2)的空间配位构建的Cu活性中心呈现最优的催化性能,以乙腈(MeCN)为溶剂,n(CuL_(2)Cl_(2)):n(AN)=1:100,100℃下,反应9 h,TBN的收率可达91.6%,且催化剂可简便回收使用4次而活性无明显降低。基于相关的实验结果,提出了CuL_(2)Cl_(2)催化CCl_(4)和AN的ATRA反应中Cu(Ⅱ)/Cu(Ⅰ)的氧化-还原催化循环机理。