Acute Toxicity of Jinchuan Formula Plum Wine Extract and Its Protective Effect on Mice with Liver Injury

[Objectives]To investigate the acute toxicity and hepatoprotective effect of Jinchuan formula plum wine extract on mice,determine its safety range,and evaluate its hepatoprotective effect.[Methods]The median lethal dose(LD_(50))was determined by acute toxicity test with the toxic reaction and mortality of mice as indexes.Sixty Kunming mice were randomly divided into 6 groups:normal control group,model group(ConA-induced liver injury model),Jinchuan formula plum wine high,medium and low dose groups(1.0,0.5,0.25 g/kg)and silybin group(0.1 g/kg).The levels of ALT,AST,LDH in serum and TG,VLDL in liver were measured.After HE staining,the pathological changes of liver tissue in mice were observed,and the liver protective effect of Jinchuan formula plum wine extract was analyzed and evaluated.[Results]LD_(50)was 11.18 g/kg,and the 95%confidence limit of LD_(50)was 10.31-12.05 g/kg.The high-dose group of Jinchuan formula plum wine extract could significantly reduce the serum ALT and AST activities of ConA-induced liver injury mice(P<0.05).[Conclusions]Jinchuan formula plum wine extract is relatively safe,and also has a protective effect on liver injury.

Preliminary study on the protective effect of electroacupuncture Neiguan acupoint pretreatment on rats with myocardial ischemia-reperfusion injury:role of the miR-214-3p/NCX1 axis

Background:Ischemia-reperfusion can worsen myocardial damage and increase the risk of death.Studies have revealed that ischemic preconditioning provides the best endogenous protection against myocardial ischemia-reperfusion injury(MIRI),and the principle of electroacupuncture(EA)preconditioning is comparable to that of myocardial ischemic preconditioning adaption.Our earlier research demonstrated that EA pretreatment inhibits the expression of calmodulin-dependent protein kinase IIδ(CaMKIIδ),sodium/calcium exchanger 1(NCX1),and cyclophilin D,hence providing protection against MIRI.However,the exact mechanism is still unknown.The expression of NCX1 mRNA is directly regulated by microRNA-214(miR-214).Moreover,it suppresses the levels of CaMKIIδand cyclophilin D.Whether these variables contribute to EA preconditioning to improve MIRI needs to be investigated,though.This study aimed to preliminarily determine whether EA pretreatment ameliorates MIRI by modulating the miR-214-3p/NCX1 axis.Methods:We used a rat MIRI model to investigate the effect of EA pretreatment on MIRI and the expression of miR-214-3p.In addition,adenovirus injection inhibited miR-214-3p expression in the rat MIRI model,and the influence of EA pretreatment towards MIRI was observed in the context of blocked miR-214-3p expression.Both the myocardial histological abnormalities and the alterations in the ST segment of the rat electrocardiogram were analyzed.NCX1 mRNA,cyclophilin D,and CaMKIIδexpression levels were also analyzed.Results:EA pretreatment improved MIRI.In rats with MIRI,EA administration increased miR-214-3p expression while decreasing NCX1 mRNA,cyclophilin D,and CaMKIIδproteins in cardiac tissues.The beneficial effect of EA pretreatment against MIRI was reversed,coupled with elevated levels of NCX1 mRNA,cyclophilin D,and CaMKIIδprotein expression,when an adenovirus injection disrupted the expression of miR-214-3p.Conclusions:Our findings preliminarily show that EA pretreatment inhibits the expression of NCX1 mRNA,cyclophilin D,and CaMKIIδproteins via miR-214-3p,hence exerting MIRI protection.

Protective effect of Solanum Nigrum Linn green fruit ethanolic extract on alcoholic liver injury in mice

Alcoholic liver injury is a liver disease caused by excessive alcohol consumption,which can lead to chronic liver disease death.Solanum Nigrum Linn taste bitter,cold,has the effect of clearing heat and detoxification,promoting blood and detumescence.Solanum Nigrum Linn fruit contains a variety of antioxidant enzymes,can remove the body produced by aerobic metabolism harmful substances.In this paper,a model of alcohol-induced liver injury in C57BL/6 mice was established to evaluate the protective effect of Solanum Nigrum Linn green fruit(SNGF)ethanolic extract on alcohol-induced liver injury.H&E staining and oil red O(ORO)staining showed that hepatic lobules were clearly demarcated,vacuoles were significantly reduced and lipid droplets were reduced in SNGF ethanolic extract treatment group.Serum levels of TC,TG,LDH,TBA,AKP,ALT and AST were decreased in the SNGF ethanolic extract treatment group,and SNGF ethanolic extract could clear reactive oxygen species(ROS)in time.MDA content was signifi cantly decreased after SNGF ethanolic extract treatment,while superoxide dismutase(SOD)and GSH-Px contents were increased after SNGF ethanolic extract treatment.These results suggest that SNGF ethanolic extract has a protective effect on alcohol-induced liver injury.

Protective Effect of Ethanol Extract from Sweet Potato Leaves on CCL_(4)-Induced Liver Injury in Mice

[Objectives]To investigate the protective effect of ethanol extract from sweet potato leaves on liver injury induced by CCl_(4)in mice.[Methods]25 ICR mice were randomly divided into blank group,model group,high-dose extract group(200 mg/kg),low-dose extract group(100 mg/kg)and positive control group(2 mg/kg colchicine),with 5 mice in each group.All groups except the blank group were given intraperitoneal injection of 20%CCl 4 olive oil solution(2 mL/kg),and the blank group was given the same dose of olive oil solution three times a week.After 4 weeks,each administration group was given the corresponding dose of drugs(10 mL/kg),and the blank group and model group were given the corresponding amount of normal saline for 2 weeks.After the last intragastric administration,fasting was required,but water was allowed,blood was taken from eyeballs,and upper serum was taken by static centrifugation.Serum AST,ALT,CRP,IL-6 and SOD levels were detected by the kit.[Results]Compared with the blank group,the serum AST and ALT levels in the model group were significantly increased;compared with the model group,the ethanol extract of sweet potato leaves could decrease the levels of ALT,AST,CRP,IL-6 and increase the level of SOD in serum.[Conclusions]The ethanol extract of sweet potato leaves had protective effect on the mice with liver injury induced by CCl_(4),and its mechanism may be to protect the liver by lowering enzymes,inhibiting inflammation and antioxidant stress.

Protective Effect of Dimethyl-4,4'-Dimethoxy-5,6,5',6'-Dimethylene Dioxybiphenyl-2,2'-Dicarboxylate (DDB) against Carcinogen-Induced Rat Liver Nuclear DNA Damage

The protective effect of DDB against carcinogen-induced DNA damage was examined in the present investigation. Preincubation of rat liver nuclei with DDB (1 mmol.L-1) resulted in 60% inhibition of binding of 3H-benzo (a) pyrene to nuclear DNA. Unscheduled DNA synthesis (UDS) induced by aflatoxin BI (10^(-7) mol.L-1) in freshly isolated rat hepatocytes was also inhibited by DDB (10^(-6)-10^(-3)mol.L-1). Oral administration of DDB at 200 mg.kg-1 once daily for 3 d induced a significant increase of liver cytosol glutathione-S-transferase and microsomal UDPG-transferase activity in mice. These results indicate that DDB is able to directly or indirectly antagonize certain carcinogen-induced DNA damages.

Protective effects of C-phycocyanin on alcohol-induced acute liver injury in mice

Excessive alcohol consumption leads to liver disease. Extensive evidence suggests that C-phycocyanin(C-PC), a chromophore phycocyanobilin derived from Spirulina platensis, exerts protective eff ects against chemical-induced organ damage. In this study, we investigated whether C-PC could protect against ethanol-induced acute liver injury. Serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), triglyceride(TG), total cholesterol(CHOL), low-density lipoprotein(LDL), liver homogenate malondialdehyde(MDA), superoxide dismutase(SOD) content were measured, and pathological examination of liver sections were examined. C-PC showed obvious inhibitory eff ects on serum ALT, AST, TG, CHOL, LDL and MDA, and SOD content significantly increased in the liver. The structure of hepatic lobules was clear, liver sinus returned to normal, and liver cell cords were arranged in neat rows. Cloudiness, swelling, inflammatory cell infiltration and spotty necrosis of liver cells were significantly reduced. Therefore, C-PC can significantly protect against ethanol-induced acute liver injury.

Late Protective Effects of the Anticalmodulin Drug Fluphenazine on Carbon Tetrachloride-induced Liver Necrosis

Fluphenazine (FP) treatment (50mg/kg bw, ip in saline) 30 min before or 6 or 10 h after CCl4 administration (1 ml/kg ip in olive oil) significantly prevented the liver necrosis produced by the hepatotoxin at 24 h. FP had enhancing effects on the covalent binding of CCl4 reactive metabolites to cellular constituents and on CCl4 induced lipid peroaldation.FP lowered bOdy temperature of the CCl4-poisoned animals during the 24 h observation period. The obtained results are compatible but do not prove the hypothesis that calmodulin (CaM) had participation in late occurring events preceding necrosis. FP lowering action on body temperature, however, might also play a role in the effects of this drug on the onset of CCl4 induced liver necrosis. FP levels in liver tissue as determined by gas chromatography-mass spectrometry evidenced the presence of the drug in amounts suffi cient to inhibit CaM and that suggests that not all preventive effects of FP are due to its indirect actions on the central nervous system via decreased body temperature

Protective effect of fu-qi granule on carbon tetrachloride-induced liver fibrosis in rats

AIM： To investigate the effcacy of fu-qi granule （FQG） on carbon tetrachloride （CCl4） induced liver fbrosis in rats and the underlying mechanisms. METHODS： Sixty rats were randomly divided into six groups： normal control group, CCl4 induced liver fbrosis group, AnluoHuaxianWan group and three treatment groups of FQG. Treatment of rats with intraperitoneal injection of carbon tetrachloride solution at 0.3 mL per 100 g body weigh twice a week for 8 wk. The normal control group the rats were given the media （olive oil） at the same time. In the frst 2 wk, rats were raised with feedstuff （80% corn meal, 20% lard, 0.5% cholesterol）. Serum samples were collected for alanine transaminase, aspartate aminotransferase, alkaline phosphatase, albumin, total protein assay and typical histopathological changes was observed in Hematoxylin-eosin staining sections. Smooth muscle alpha actin （α-SMA） was analyzed with immunohistochemistry. Mammalian target of rapamycin （mTOR） and hypoxia-inducible factor-1 （HIF-1α） expressions were detected by Western blot-ting. Tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） and matrix metalloproteinases-9 （MMP-9） were measured with semi-quantitative reverse transcriptase-polymerase chain reaction.RESULTS： FQG significantly reduced the serum levels of alanine transaminase, aspartate aminotransferase, alkaline phosphatase and increased the serum contents of albumin, total protein in rats with liver fibrosis. Moreover, FQG promoted extracellular matrix degradation by increasing MMP-9 and inhibiting TIMP-1 and α-SMA. mTOR and HIF-1α expression in liver significantly decreased in the rats treated with FQG. CONCLUSION： The results indicated that FQG signi-fcantly reverse fbrosis induced by CCl4, which should be developed as a new and promising preparation for the prevention of liver fbrosis.

Protective Effect of Vitamin E on Liver Damage Induced by 2-Chloro-1, 3-butadiene

The present study was performed to determine the influence of lipid peroxidation and perturbance of Ca2+ homeostasis on liver damage induced by 2-chloro-1, 3-butadiene (CBD) and the protective effects of vitamin E in Wistar rats. Animals were given intraperitoneally different doses (8,40 or 200 mg·kg-1 daily) of CBD for 21 days, and the following dose-dependent events were observed: liver damage, significant increase in liver lipid peroxides, and decreases in activities of erythrocytic glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). The pretreatment of rats with vitamin E (po 150 mg·kg-1) before administering CBD (iP 60 mg·kg-1 ) daily for 21 days prevented the following CBD-induced changes, the increase in serum cholylglycine (CG), hepatic LP, hepatic mitochondrion LP, hepatic oxidized glutathione (GSSG) (while the significant increase of reduced glutathione (GSH) was not affected) and the decrease in activities of erythrocytic SOD and hepatic mitochondrial calcium sequestration. These results suggest that lipid peroxidation and perturbance of Ca2+ homeostasis appear to contribute to the hepatotoxicity of CBD, and vitamin E might prevent the liver damage induced by CBD. The decrease in activities of GSH-Px and SOD in erythrocytes might be used as biomarkers for adverse effects of CBD on defense system against lipid peroxidation.

Protective effect of hydrogen-rich saline on liver ischemia-reperfusion injury in mice

Objective To explore protective effect of hydrogen - rich saline on liver ischemia reperfusion ( IR) in mice and possible mechanisms. Methods Twenty - four C57BL /6 mice were randomly divided into 3 groups: sham - operated group,control group ( mice were injec-

Effects of natural cerebrolysin on protective proteins and pro-apoptotic molecules in mesenchymal stem cells following beta-amyloid peptide1-40-induced endoplasmic reticulum stress

BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mechanisms of traditional Chinese medicine against ERS in AD are poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. NC was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yinxingye （Ginkgo Leaf） in a proportion of 1 ： 2： 2. Following conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.

The Extraction of Eucommia Ulmoides Oliv Polysaccharides and Its Protective Effect on Liver of Clophasphamidecy Injured Mice

Objective:To study the influenceof eucommia polysaccharide on the mice' liver damaged by clophasphamidecy (CY).Methods:Injecting CY build mice liver damage model,eucommia polysaccharide given different doses, measured blood serum ALT,AST and the liver's SOD,MDA. Results:After the injection CY,blood serum ALT,AST and the MDA of liver rise and the SOD of liver reduce comparedwith the blank group. The eucommia polysaccharide can improve these index.Conclusion:The Eucommia polysaccharide may protect the mice' liver damaged by CY.

Protective Effect of Mongolian Medicine Youning Bawei Powder on Liver Injury Induced by CCl4 in Mice

[Objectives]To study the protective effect of Mongolian medicine Youning Bawei Powder on liver injury induced by carbon tetrachloride(CCl_(4))in mice.[Methods]The experimental mice were divided into 5 groups:normal group,model group,positive control group,low-dose group and high-dose group of Mongolian medicine Youning Bawei Powder.The model group was induced by CCl_(4),the positive control group was treated with liver-protecting tablet,and the Mongolian medicine group was treated with Youning Bawei Powder for one month.The liver tissue injury of mice in each group was observed by HE staining,and the levels of serum ALT,AST,SOD and MDA were detected.[Results]Mongolian medicine Youning Bawei Powder significantly improved the pathological liver injury induced by CCl_(4),significantly decreased the content of ALT,AST and MDA in serum of mice with CCl_(4) liver injury,and significantly increased the activity of serum SOD.[Conclusions]Youning Bawei Powder,a Mongolian medicine,has a protective effect on liver injury induced by CCl_(4) in mice.

LXR及其靶基因COX-2和CETP在肥胖OSAHS幼鼠肝组织中的保护作用

目的阐明肝X受体(liver X receptor,LXR)及其靶基因环氧化酶-2(cyclooxygenase-2,COX-2)、胆固醇酯转移蛋白(cholesteryl ester transfer protein,CETP)的高表达是肥胖幼鼠阻塞性睡眠呼吸暂停综合征(obstructive sleep apnea-hypopnea syndrome,OSAHS)发病过程中的保护性因素,为肥胖儿童OSAHS的发病机制提供基础研究资料。方法24只3~4周龄雄性Wistar幼鼠分为正常对照组(control组)、单纯肥胖组(obesity组)、单纯OSAHS组(OSAHS组)、肥胖+OSAHS组(obesity+OSAHS组)。HE染色观察幼鼠肝组织病理变化;蛋白免疫印迹法(Western blotting)检测幼鼠肝组织中LXRα、COX-2、CETP的表达水平;运用免疫组化方法检测幼鼠肝组织中LXRα、COX-2、CETP的表达水平及分布情况。结果单纯肥胖组和肥胖+OSAHS组幼鼠体质量、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)含量与正常对照组相比均明显增加(P<0.05),单纯OSAHS组和肥胖+OSAHS组幼鼠血氧饱和度与正常对照组相比均明显降低(P<0.05)。单纯肥胖组、单纯OSAHS组及肥胖+OSAHS组肝组织与正常对照组肝组织相比均有明显损伤,肥胖+OSAHS组肝组织损伤较单纯肥胖组、单纯OSAHS组肝组织损伤程度明显升高。单纯OSAHS组和单纯肥胖组幼鼠肝组织中LXRα、COX-2、CETP表达水平较正常对照组均明显升高(P<0.05)。肥胖+OSAHS组幼鼠肝组织中LXRα、COX-2、CETP表达水平较其余各组均明显升高(P<0.05)。结论LXR及其靶基因COX-2、CETP在肥胖OSAHS幼鼠肝脏中高表达,是发病过程中的可能保护性因素。

小分子多肽LK-5对CCl4致急性肝损伤小鼠的保护作用研究

【目的】研究小分子多肽LK-5(氨基酸序列为LHMFK)对CCl4致急性肝损伤模型小鼠的保护作用,探讨其作用机制。【方法】72只小鼠随机分为正常组、模型组、联苯双酯组(150 mg/kg)及LK-5低、中、高剂量组(30、60、120 mg/kg),每组12只。连续给药10 d, 1次/d,每次10 mL/kg。末次给药2 h后,各组(除正常组外)腹腔注射0.1%CCl4花生油溶液,建立CCl4致小鼠急性肝损伤模型,16 h后,按照试剂盒方法,检测小鼠血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、总胆汁酸(TBA)和碱性磷酸酶(AKP)水平,检测肝脏超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、肿瘤坏死因子(TNF-α)、白细胞介素-6(IL-6)、IL-10、血管紧张素1-7(Ang 1-7)和血管紧张素Ⅱ(AngⅡ)水平。制作肝脏病理组织切片,苏木精-伊红染色(HE)后观察肝脏组织病理学变化。【结果】与正常组比较,模型组小鼠肝细胞有明显的脂肪变性,炎细胞分散分布于肝小叶血管周围和肝实质内,血清ALT、AST、AKP活性和TBA水平均显著上升,肝组织SOD、GSH-Px活性显著下降,MDA水平显著上升(P<0.05),说明成功建立急性肝损伤模型,TNF-α、IL-6、AngⅡ水平显著升高(P<0.05)。与模型组比较,LK-5中、高剂量组能够显著降低血清ALT、AST活性(P<0.05),各剂量组均可降低血清TBA水平和AKP活性(P<0.05),LK-5高剂量组能够显著提高小鼠肝组织SOD、GSH-Px活性(P<0.05),LK-5中、高剂量组能够显著降低肝组织MDA水平(P<0.05),LK-5高剂量组TNF-α水平显著降低(P<0.05),各剂量组能够显著降低肝组织IL-6、AngⅡ水平(P<0.05)。病理切片结果显示,LK-5各剂量组炎细胞均有不同程度减少,LK-5高剂量组肝小叶血管周围和肝实质内无炎细胞浸润。【结论】LK-5各剂量组对CCl4致急性肝损伤小鼠具有保护作用,其中60、120 mg/kg LK-5效果更为明显,其作用机制可能与抗氧化、抗炎及对肾素-血管紧张素系统(RAS)的调控有关。

枸杞多糖对肝内胆汁淤积模型大鼠肝脏预防保护作用的研究

目的 探讨枸杞多糖(LBP)对肝内胆汁淤积模型大鼠肝脏损伤的预防保护作用。方法 SPF级雄性SD幼龄大鼠,设阴性对照组、模型组、LBP 50、150、450 mg/kg剂量组,以及阳性对照组,每组各10只。构建大鼠肝内胆汁淤积模型,模型组、LBP各剂量组及阳性对照组,予60mg/kg α-萘异硫氰酸酯,每隔1天给药一次,第2周开始改为每隔3天给药一次,持续6周。第2周起,各剂量组每天予不同剂量的LBP,阳性对照组予熊去氧胆酸,模型组大鼠予蒸馏水。6周后采集血液及肝组织样本,测定血清生化指标、凝血功能指标及白细胞分类计数指标,检测肝组织中的炎症因子含量以及Toll样受体4(TLR4)、核因子κB(NF-κB)、核转录因子(pNF-κB)及髓分化因子88(MyD88)蛋白的表达量,观察大鼠肝脏的病变情况。结果 与阴性对照比较,模型组大鼠血清γ-谷氨酰转移酶(γ-GGT)、碱性磷酸酶(ALP)、总胆汁酸(TBA)、总血红素(TBiL)、胆固醇(CHOL)、丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)水平,血液白细胞(WBC)、淋巴细胞(LYMP)、中性粒细胞(NEUT)、单核细胞(MONO)及嗜酸性粒细胞(EO)计数,纤维蛋白原(FIB)含量均升高(均P <0.05),肝组织中炎症因子肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)、白介素6及巨噬细胞炎性蛋白2(MIP-2)含量增加,TLR4、pNF-κB、MyD88蛋白表达水平上调(均P <0.05)。肝脏可见多量炎症细胞浸润、胆管增生、纤维组织增生等病理性改变,组织形态学观察评分上升(均P <0.05);与模型组比较,LBP450mg/kg组大鼠血清γ-GGT、TBA、TBiL、ALT水平,血液WBC、LYMP、NEUT计数,FIB含量均下降(均P<0.05),肝组织中炎症因子TNF-α、IL-1β、MIP-2含量降低,TLR4、pNF-κB、MyD88蛋白的表达水平下调(均P<0.05),肝脏组织形态学观察评分下降(均P <0.05),肝脏损伤减轻,LBP 150 mg/kg组大鼠血液FIB含量均下降,TLR4蛋白的表达水平下调(均P<0.05)。结论 LBP对肝内胆汁淤积模型大鼠肝脏损伤有缓解和保护功效,其作用机制可能与下调TLR4/MyD88/NF-κB信号通路相关蛋白的表达,抑制过度炎症反应有关。

陈皮复合固体饮料制备工艺及其对急性酒精性肝损伤小鼠的保护作用

[目的]以药食同源的陈皮、山楂、葛根为主要原料,开发一种具有解酒功能的固体饮料,并探究其对急性酒精性肝损伤小鼠的保护作用。[方法]以浸膏得率和总黄酮含量为标准,采用正交试验选取最佳提取工艺;以感官评价、成型率、溶化性的综合评分为指标,采用单因素试验优化成型工艺。采用52%酒精灌胃的方法建立急性酒精性肝损伤小鼠模型,检测陈皮复合固体饮料对模型小鼠肝脏指数,血清中谷草转氨酶(AST)、谷丙转氨酶(ALT),肝组织中总胆固醇(TC)、甘油三酯(TG)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)及炎症因子白细胞介素-1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)的影响,苏木素—伊红(HE)染色分析各组肝脏病理变化。[结果]陈皮复合固体饮料最佳制备工艺:3味药材浸泡30 min后提取2次,第一次料液比1∶12(g/mL)、提取时间2.0 h,第二次料液比1∶10(g/mL)、提取时间1.5 h;经浓缩、干燥、粉碎后得到干膏粉;干膏粉与麦芽糊精1∶1混合,加入1.0%的罗汉果甜苷,并以90%乙醇作为润湿剂进行湿法制粒。动物试验表明:与模型组相比,陈皮复合固体饮料组的AST、ALT、TG、TC、MDA、IL-1β、IL-6、TNF-α水平降低(P<0.01或P<0.05),GSH、SOD含量增加(P<0.01或P<0.05),且肝组织病理切片显示给药组小鼠肝损伤程度均有显著改善。[结论]陈皮复合固体饮料色泽均匀、溶解性好、甜度适中、口感细腻,对急性酒精性肝损伤具有良好的保护作用。

水飞蓟宾-磷脂酰胆碱复合物对四氯化碳致小鼠急性肝损伤的保护作用

目的:研究水飞蓟宾-磷脂酰胆碱复合物(SPC)对四氯化碳(CCl4)所致的小鼠急性肝损伤的预防和治疗作用。方法:50只小鼠随机分成5组,每组10只:正常对照组、模型组、预防组、治疗组和预防+治疗组,采用CCl4造成小鼠急性肝损伤模型,预防组于注射CCl4前多次给小鼠灌服SPC,治疗组于注射CCl4后给SPC,预防+治疗组于注射CCl4前后均给SPC,24h后采血,分离血清,检测血清ALT和AST活性,并取肝组织观察比较肝脏病理组织学改变。结果:SPC明显降低小鼠血清ALT和AST活性,治疗、预防及预防+治疗组与模型组比较差异均有极显著性(P<0.01),预防组与治疗组比较差异有显著性(P<0.05),预防+治疗组与治疗组比较差异有极显著性(P<0.01),且均能改善肝脏病理组织学变化。结论:SPC对CCl4造成的急性肝损伤有明显的预防和治疗作用。

灵五颗粒对D-半乳糖胺盐酸盐致小鼠肝损伤的保护作用

目的观察灵五颗粒对D-半乳糖胺盐酸盐致小鼠急性肝损伤的保护作用。方法将180只小鼠随机分为正常组、D-半乳糖胺盐酸盐模型组、灵五颗粒低、中、高(2.5、5.0、10g/kg)剂量组和阳性药联苯双酯(150mg/kg)组。除正常组、D-半乳糖胺盐酸盐模型组给予与灵五颗粒高剂量同等剂量的生理氯化钠溶液外,给药各组均分别给药7d,于末次给药1h后,以D-半乳糖胺盐酸盐(800mg/kg)给予各组小鼠(正常组除外)腹腔注射中毒,造成急性肝损伤,在中毒后12h各给药组再给药1次,于中毒24h杀鼠取血和肝脏,分别检测各组动物肝功能等指标,并取肝脏作病理组织学检查。结果 D-半乳糖胺盐酸盐模型组小鼠肝功能明显异常,谷丙转氨酶、谷草转氨酶显著升高,肝组织变性、坏死严重;灵五颗粒中、高剂量组小鼠血清中谷丙转氨酶的水平均非常显著低于模型组,并呈良好的剂量依赖性。灵五颗粒高剂量组小鼠血清中谷草转氨酶的水平显著低于模型组,此外,还可显著降低损伤小鼠丙二醛的含量,显著升高其超氧化物歧化酶活力。结论灵五颗粒对D-半乳糖胺盐酸盐致小鼠急性肝损伤有明显的保护作用。

可溶性mB7-H4对Con A诱导小鼠肝损伤的保护作用

目的:探讨可溶性mB7-H4蛋白对肝脏的免疫性损伤的保护作用,阐明其作用机制。方法:采用异丙基硫代半乳糖苷(IPTG)诱导pET/mB7-H4表达,通过纯化、复性以及去除内毒素,获得具有生物活性的可溶性mB7-H4蛋白。小鼠随机分为对照(生理盐水)组、Con A组和mB7-H4蛋白100、200及400μg组,除对照组外其余各组小鼠通过尾静脉注射Con A建立肝损伤模型(mB7-H4蛋白组小鼠在注射Con A前2h和注射后8h分2次给予mB7-H4蛋白),并于注射Con A后24和48h,摘取各组小鼠眼球取血及引颈处死取肝脏,分离血清,检测天冬氨酸氨基转移酶(ALT)、丙氨酸氨基转移酶(AST)、白细胞介素4(IL-4)和γ干扰素(IFN-γ)水平,并检测肝脏组织病理学变化。结果:与对照组比较,其他各组小鼠血清ALT、AST、IL-4和IFN-γ水平较均明显升高(P<0.05);与Con A组比较,mB7-H4蛋白组小鼠血清ALT、AST、IL-4和IFN-γ水平均明显降低(P<0.05或P<0.01)。HE染色,mB7-H4蛋白组小鼠肝脏损伤均较Con A组有不同程度的减轻。结论:可溶性mB7-H4蛋白对Con A诱导的肝脏免疫性损伤具有保护作用,可能是通过抑制IL-4和IFN-γ的产生和(或)分泌而发挥作用。