Effect of compound rhodiola sachalinensis A Bor on CCI4-induced liver fibrosis in rats and its probable molecular mechanisms

AIM: To explore the anti-fibrotic effect of a traditional Chinese medicine, compound rhodiola sachalinensis A Bor on CCl4-induced liver fibrosis in rats and its probable molecular mechanisms. METHODS: Ninety healthy male SD rats were randomly divided into three groups: normal group (n=-10), treatment group of compound rhodiola sachalinensis A Bor (n=-40) and CCl4-induced model group (n=40). The liver fibrosis was induced by CCl4 subcutaneous injection. Treatment group was administered with compound rhodiola sachalinensis A Bor (0.5 g/kg) once a day at the same time. Then the activities of several serum fibrosis-associated enzymes: alanine aminotransferase (ALT), aspartate aminotransferase (AST), N-acetyl-beta-D-glucosaminidase (β-NAG) and the levels ofserum procollagen Ⅲ (PCⅢ), collagen Ⅳ (CⅣ), hyaluronic acid （HA） were assayed. The histooathol(mical chanaes were observed with HE, VG and Masson stain. The expression of TGF-β1 mRNA,αl (I) mRNA and Na+/Ca2+ exchanger (NCX ) mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) in situ.RESULTS: Compound rhodiola sachalinensis A Bor significantly reduced serum activities of ALT, AST, β-NAG and decreased the levels of PCⅢ, CⅣ, HA, improved the liver histopathological changes, inhibited the expression of TGF-β1 mRNA, α(1) mRNA and Na+/Ca2+ exchanger mRNA in rats. CONCLUSION: Compound rhodiola sachalinensis A Bor can intervene in CCI4-induced liver fibrosis in rats, in which potential mechanisms may be decreasing the production of TGF-β1, reducing the production of collagen, preventing the activation of hepatic stellate cell (HSC) and inhibiting theexpression of TGF-β1 mRNA, αl(I) mRNA and Na+/Ca2+ exchanger mRNA.

依那普利对CCI_4急性肝损伤大鼠抗氧化功能的影响

目的:研究依那普利对四氯化碳(CCl4)所致急性肝损伤大鼠肝损伤和抗氧化功能的作用. 方法:将50只♂SD大鼠随机分为5组(每组10只):药物干预组(10mg/kg,5mg/kg和2.5mg/kg)、模型组和正常对照组,药物干预组和模型组均给予皮下注射CCl4(用等体积的橄榄油稀释),以制备急性肝损伤的大鼠模型,正常对照组用等体积的生理盐水注射,用高(10 mg/kg)、中(5 mg/kg)、低(2.5 mg/kg)剂量依那普利分别对SD大鼠灌胃给药.全自动生化分析仪测定大鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、碱性磷酸酶(ALP)、胆汁酸(TBA)的活性;用比色分析法测定超氧化歧化酶(T-SOD)、黄嘌呤氧化酶(XOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性及丙二醛(MDA)的含量. 结果:依那普利显著降低因CCl4所致急性肝损伤大鼠血清ALT(正常对照组685±63 nkat/L<10 mg干预组1 241±168 nkat/L<5 mg干预组1 705±83 nkat/L<2.5 mg干预组2 302±174 nkat/L<模型对照组3 531±776 nkat/L),AST (正常对照组1 240±158nkat/L<10 mg干预组2 430±386 nkat/L<5 mg干预组2 788±522 nkat/L<2.5 mg干预组3 151±917 nkat/L<模型对照组3 372±138 nkat/L), ALP(10mg干预组2 567±159nkat/L<正常对照组2 659±248 nkatL<5 mg干预组3 212±198 nkat/L<2.5 mg干预组3 231±261 nkat/L<模型对照组3 609±346 nkat/L)和TBA(正常对照组8.48±0.49 μmol/L<10 mg干预组16.35±5.43μmol/L<5 mg干预组16.92±2.68 μmol/L<2.5 mg干预组17.53±3.59μmol/L<模型对照组24.16±9.27μmol/L) 的升高.对急性肝损伤大鼠血清GSH-PX(模型对照组50±54 nkat/L<2.5 mg干预组149±111 nkat/L<10 mg干预组169±141 nkat/L<5 mg干预组1 70±91 nkat/L<正常对照组295±194 nkat/L)的活性有明显的升高作用及降低T SOD(正常对照组6006±639 μkat/L<10mg干预组7 135±1 560μkat/L<2.5 mg干预组7 538±938 μkat/L<5 mg干预组7 589±780μkat/L<模型对照组8 579±861μkat/L) 和XOD(正常对照组571±28 nkat/L<10 mg干预组724±18nkat/L<5mg干预组821±28nkat/L<2.5mg干预组868±58 nkat/L<模型对照组1 042±188nkat/L)的含量.以上均与模型对照组比较aP<0.05,bP<0.01. 结论:依那普利的保肝机制和抗氧化作用与其对抗自由基脂质过氧化密切相关.

番茄红素对CCl_4致大鼠慢性肝损伤的保护作用

目的探讨番茄红素对慢性肝损伤的保护作用。方法将40只SD大鼠随机分为正常对照组、单纯肝损伤组及番茄红素低、中、高剂量组,采用CCl4制备慢性肝损伤大鼠模型,检测大鼠血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)活性及肝组织匀浆中超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和丙二醛(MDA)活性,同时对肝脏进行病理学评估。结果番茄红素能明显降低慢性肝损伤模型大鼠血清中AST、ALT活性(P<0.01),降低肝组织匀浆中MDA活性(P<0.01),升高GSH和SOD活性(P<0.01),降低肝脏病理损伤程度。结论番茄红素对CCl4所致的大鼠慢性肝损伤具有一定的保护作用。

脂糖舒对CCl_4所致大鼠肝损伤的保护作用

目的探讨脂糖舒对实验性损伤的保护作用。方法建立大鼠CCL4 肝损伤模型 ,将大鼠随机分为正常组、模型组、治疗组。治疗组于建模后灌胃施药。检测丙氨酸转氨酶 (ALT) ,观察肝脏病理学变化。结果脂糖舒可以抑制CCl4 导致的大鼠血清ALT升高 ,使肝细胞变性、坏死得到明显的改善和恢复。

中药保肝康抗CCl_4慢性肝损伤的实验研究

目的：研究中药保肝康抗慢性肝损伤的作用及机理。方法：采用ＣＣｌ４ 复制慢性肝损伤动物模型，设甘草甜素为对照药物。结果：保肝康具有改善肝功的作用，并可以改善肝组织病理；在对慢性肝损伤减轻脂肪变性、阻止肝纤维化方面均优于甘草甜素。

丹参酮Ⅱ_A对小鼠肝损伤的保护作用

目的:探讨丹参酮ⅡA(tanshinone Ⅱ A,TSN)对肝脏损伤的保护作用。方法:CCl_4和D-半乳糖胺小鼠肝脏损伤模型,以测定血清谷丙氨酸转换酶(ALT)、谷草氨基酸转换酶(AST)和肝匀浆丙二醛(MDA)含量为指标观察丹参酮ⅡA对肝脏损伤的保护作用。结果:丹参酮ⅡA与阳性对照药联苯双酯相仿,均能明显降低急性肝损伤小鼠的血清ALT、AST和肝匀浆MDA含量。结论:丹参酮ⅡA具有明显的保肝作用。

水蓑衣提取物对CCl_4诱导的小鼠急性肝损伤的保护作用

目的:探讨水蓑衣提取物对肝脏的保护作用方法:40只雄性昆明种小鼠,随机分为对照组、CCl4损伤对照组、低剂量水蓑衣(45g/kg)组、高剂量水蓑衣(90g/kg)组和联苯双酯(150mg/kg)组.除正常对照组外,其余动物均采用ipCCl4复制急性肝损伤小鼠模型,CCl4注射后16h处死动物,测定各组小鼠血清ALT、AST的活性以及检查肝组织的病理改变,观察不同剂量水蓑衣对肝脏的保护作用.结果:与正常对照组相比,CCl4损伤对照组动物血清ALT和AST活性显著升高fP<0.01).肝脏出现轻度、中度和重度坏死及变性,低剂量水蓑衣组、高剂量水蓑衣组血清ALT和AST明显低于CCl4损伤对照组(ALT:1485±755,1211±528vs3179±106;AST:2045±293,2052±386vs2583±116,P<0.01),与联苯双酯组相比无显著差异.水蓑衣能明显减轻CCl4引起的肝脏结构损害,高剂量水蓑衣的作用尤为突出,其作用优于联苯双酯.结论:水蓑衣呈剂量依赖性地减轻CCl4引起的肝损害,其作用优于联苯双酯.

蛹虫草多糖对四氯化碳诱导大鼠原代肝细胞损伤的保护作用研究

蛹虫草是一种药用真菌。从人工培养的蛹虫草中提取蛹虫草多糖(CMPS),采用Ⅳ型胶原酶灌流法分离大鼠肝细胞进行原代培养,研究蛹虫草多糖对CCl4诱导大鼠原代肝细胞损伤的保护作用。结果表明:蛹虫草的子实体和基质都含有丰富的多糖,其中基质中的多糖含量最高,约为子实体的2～4倍;CMPS可明显降低由CCl4诱导的损伤肝细胞培养上清液中谷草转氨酶(AST)、谷丙转氨酶(ALT)的活性水平和丙二醛(MDA)的含量水平,显著提高由CCl4诱导的损伤肝细胞培养上清液中谷胱甘肽过氧化物酶(GSH-Px)活性和诱导损伤肝细胞的存活率。试验结果提示CMPS对大鼠原代培养肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关。

桔梗皂苷-D对大鼠肝纤维化的干预作用及部分机制研究

目的:研究桔梗皂苷-D(PD)对大鼠肝纤维化的干预作用及作用机制。方法:将大鼠随机分为5组,除对照组外,其余各组每周2次皮下注射四氯化碳(CCL4)溶液诱导肝纤维化模型,次日PD低剂量组、PD中剂量组、PD高剂量组分别给予PD 10、25、50 mg·kg-1·d-1灌胃持续给药治疗,对照组和模型组以等量0.9%氯化钠溶液代替。给药12周后,处死大鼠采集样本,检测各组大鼠血清ALT、AST、TNF-α、血清肝纤维指标(HA、LN、PIIIP、C-Ⅳ)水平;肝组织转化生长因子-β1(TGF-β1)、骨形成蛋白-7(BMP-7)、α-平滑肌肌动蛋白(α-SMA)mRNA的表达量;HE和天狼猩红染色观察肝组织病理学变化。结果:与模型组比较:PD各组能显著降低大鼠血清ALT、TNF-α、HA及肝组织TGF-β1mRNA水平,显著升高肝组织BMP-7mRNA水平(P<0.05);PD中剂量组和高剂量组能显著降低大鼠血清AST、LN及肝组织α-SMA mRNA水平(P<0.05);PD高剂量组能显著降低大鼠血清PIIIP、C-Ⅳ水平(P<0.05);肝组织纤维化程度明显改善。结论:PD具有抗肝纤维作用,并可能是通过降低TGF-β1和调高BMP-7的表达、抑制肝星状细胞的细胞增殖和活化,以达到抗纤维化的作用。

β-catenin在四氯化碳诱导的小鼠肝纤维化过程中的定位、表达及意义

目的探讨肝纤维化发生过程中β-连环蛋白(β-catenin)定位、表达及意义。方法健康雄性昆明小鼠(n=45)随机分为对照组(n=15)与实验组(n=30)。实验组小鼠皮下注射50%四氯化碳(CCl4)-粟米油混合液(6ml/kg),2次∕周,对照组皮下注射同等剂量的粟米油。各组分别于造模1周、4周和8周后取小鼠肝脏,常规制作石蜡切片,Masson染色及Desmin免疫组织化学法观察比较不同组别小鼠肝纤维化病理变化,RT-PCR及免疫组织化学方法检测不同时间点各组小鼠肝组织内β-catenin的表达及定位。结果 Masson染色结果显示,对照组肝汇管区结缔组织内及血管壁有少量细小纤维,实验组小鼠肝脏汇管区和中央静脉及其周围胶原纤维增多;随损伤时间延长纤维增生愈加明显。Desmin免疫组织化学结果显示,各组别均有阳性表达,但损伤组各时间点desmin阳性表达细胞数明显高于对照组(P<0.05或P<0.001)。RT-PCR结果显示,CCl4损伤1周后肝内β-cateninmRNA水平与对照组相比无明显差别(P>0.05),损伤4周及8周β-catenin mRNA水平则明显下降,与对照组相比差异均有显著性(P<0.01)。免疫组织化学结果显示,对照组β-catenin弱表达于肝细胞膜及胆管上皮细胞膜和胞质,而损伤组β-catenin阳性反应主要定位于肝内增生的细胞团及新生胆管上皮细胞质,各组间积分吸光度值差别有显著性(P<0.01)。结论 CCl4诱导的肝纤维化过程中β-catenin mRNA表达与蛋白表达不同步,阳性表达细胞主要为新生的细胞团及胆管上皮细胞。

Galectin-3在CCl_4致肝损伤及对GRP78调控的作用

目的:探讨Galectin-3对CCl_4致急性肝损伤时GRP78的调控作用.方法:选择ICR系的♂Galectin-3基因敲除型[Gal-3(-/-)]和其野生型[Gal(+/+)1小鼠,一次灌胃给予CCl_4,观察给药后10,24,48和72 h的肝组织病理改变和检测其血清谷草转氨酶(AST)和谷丙转氨酶(ALT)活性,并检测肝细胞不同细胞组分中的GRP78蛋白表达.结果:与Gal(+/+)型鼠比较,CCl_4对Gal-3(-/-)型鼠的肝组织病理损伤出现时间早、损伤程度重.Gal-3(-/-)型鼠在CCl_4灌胃后10和24 h的血清ALT活性(1 860±191 U/L vs 1356±177 U/L,t=6.12,P<0.01;2789±236 U/L vs 2468±221 U/L,f=3.14,P<0.01)及CCl_4灌胃后10 h的血清AST活性(946±89 vs 623±73 U/L,t=8.87,P<0.01)与Gal(+/+)型鼠比较显著升高.对照组中Gal-3(+/+)型鼠的微粒体组分(Mic)中的GRP78蛋白表达显著高于Gal-3(-/-)型鼠(140.9±21.1 vs 76.1±9.5,t=8.86,P<0.01).在CCl_4 ig后24 h,Gal-3(+/+)型鼠的肝细胞线粒体组分(Mt)和Mic中的GRP78蛋白表达明显升高,并显著高于Gal-3(-/-)型鼠(Mt:127.0±18.8 vs 49.1±6.3,P<0.01;Mic:166.5±23.4 vs 87.7±11.6,P<0.01).结论:Galectin-3蛋白在CCl_4致急性肝损伤中可能具有一定的保护作用.上调Mic和Mt中的GRP78蛋白表达可能是Galectin-3在CCl_4对肝细胞损伤中发挥保护作用的一个途径.

丹参滴丸联合苦参素治疗大鼠肝纤维化的实验研究

目的观察丹参滴丸联合苦参素对大鼠肝纤维化的保护作用。方法 60只大鼠随机分成3组,每组20只。对照组不制作模型,模型组和实验组采用CCI4四复合因素法诱导肝纤维化大鼠模型。各组每天给药1次,对照组和模型组给予蒸馏水灌胃;实验组给予丹参滴丸联合苦参素治疗,3组连续给药7周。分别于治疗前、治疗后检测各组大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、超氧化物歧化酶(SOD)活性及血清总蛋白(TP)、白蛋白(ALB)、丙二醛(MDA)透明质酸(HA)、层粘蛋白(CN)、Ⅲ型前胶原(PCⅢ)和Ⅳ型胶原(CⅣ)含量水平。结果丹参滴丸联合苦参素能明显抑制肝纤维化大鼠血清中ALT、AST、SOD活性的升高,增加TP和ALB含量,降低MDA、HA、CN、PCⅢ和CⅣ含量。结论丹参滴丸联合苦参素有明显的抗肝纤维化作用。

Protective effects of 5,4'-dihydroxy-3',5'- dimethoxy-7-O-β-D -glucopyranosyloxy-flavone on experimental hepatic injury

AIM: To investigate the pharmacological effects of rice flavone (5,4'-dihydroxy-3',5'-dimethoxy-7-0-β-D-glucopyranosyloxy-flavone, RF) separated from panicle-differentiating to flowing rice on rat experimental hepatic injury. METHODS: Models of rat acute hepatic injury induced by carbon tetrachloride (CCl4) administration, rat hepatic fibrosis induced by thioacetamide, injury of primary cultured rat hepatocytes induced by CCl4, respectively, were established. After treated with RF, content of serum alanine transaminase (ALT), aspartate aminotransferase (AST) and albumin (Alb), hyaluronic acid (HA), the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and hydroxyproline (Hyp) were measured and liver tissue was observed pathologically by hematoxylin-eosin (HE) staining. Effects of RF on pathological changes, function index, enzyme of scavenging free radicals and blood rheology were evaluated. RESULTS: In model of rat acute hepatic injury induced by CCI4, RF can significantly decrease the contents of serum ALT, AST, increase the content of Alb, improve the dropsy and fat denaturalization of hepatocytes. In model of rat hepatic fibrosis induced by thioacetamide, RF can inhibit the increase of HA, Hyp and whole blood viscosity, and improve the activities of GSH-Px and SOD, and inauricular microcirculation. CONCLUSION: RF has apparent protective effects on hepatic injury by increasing activity of GSH-Px and SOD, scavenging free radicals produced by CCI4, reducing blood viscosity, and improving microcirculation and blood supply.

Modifi cation of sleep architecture in an animal model of experimental cirrhosis

AIM： To analyze the polygraphic sleep patterns during cirrhosis progression in a rat model by repeated CCh administration. METHODS： Male Wistar rats received three weekly injections of CCl4 for 11 wk, and were analyzed before and during the induction of cirrhosis. Rats were im- planted with electrodes to record their sleep patterns. Polygraph recordings were made weekly over 11 wk for 8 h, during the light period. After a basal recording, rats received three weekly injections of CCl4. Histological confirmation of cirrhosis was performed after 11 wk. RESULTS： The results showed a progressive decrease in total wake time that reached statistical significance from the second week of treatment. In addition, there was an increase in total time of slow wave sleep （SWS）Ⅱ and rapid eye movement sleep （REM sleep） in most of the 11 wk. SWS I showed no significant variations. During the final weeks, a significant increase in REM sleep frequency was also observed. Histological analyses of the livers showed unequivocal signs of cirrhosis. CONCLUSION： These data suggest that hepatic failure produced by CCh administration is capable of modifying the sleep pattern even after only a few doses.

肝乐宁粉针剂对实验性肝损伤的保护作用

研究肝乐宁粉针剂对实验性肝损伤的保护作用。方法：采用四氯化碳致小鼠和大鼠肝脏损伤，测定生化指标及观察病理组织学改变。结果：肝乐宁粉针剂能显著降低四氯化碳引起的急性肝损伤小鼠和慢性肝损伤大鼠血清丙氨酸氨基转换酶和天门冬氨酸氨基转换酶增高；亦能明显降低大鼠血清唾液酸和肝羟脯氨酸含量，升高血清总蛋白和白蛋白水平；病理组织学检查肝乐宁粉针剂明显减轻肝细胞的变性和坏死，并抑制慢性肝损伤胶原纤维形成。结论：肝乐宁粉针剂具有肝细胞保护及抗肝纤维化作用。

健肝灵胶囊对小鼠肝损伤模型的保护作用

目的探讨健肝灵胶囊对小鼠免疫性肝炎及急性肝损伤保护作用。方法小鼠静脉注射卡介苗5×106个菌/鼠,11天后再静脉注射脂多糖7.5μg/鼠,造成免疫性肝炎模型;另分别用四氯化碳和硫代乙酰胺造成小鼠急性中毒模型;观察小鼠血清谷丙转氨酶、谷草转氨酶、超氧化物歧化酶、丙二醛和一氧化氮的变化,评价健肝灵胶囊对免疫性肝炎及四氯化碳和硫代乙酰胺致急性肝损伤小鼠的保护作用。结果健肝灵胶囊能明显降低免疫性肝炎及急性肝损伤模型小鼠血清谷丙转氨酶、谷草转氨酶(P<0.01),超氧化物歧化酶的活性升高,丙二醛和一氧化氮含量明显降低。结论健肝灵胶囊对小鼠免疫性肝炎及急性肝损伤有保护作用。

番茄红素对CCl_4诱导的正常肝细胞株HL-7702损伤的影响

目的体外观察番茄红素对人正常肝细胞株HL-7702细胞活力及凋亡的影响,探讨其对肝细胞的保护作用。方法体外培养人正常肝细胞株HL-7702,建立CCl4肝损伤模型,终浓度为2、5、10μmol·L-1的番茄红素干预处理,MTT法测定细胞活性、Horchest33342染色及TUNEL检测观察细胞凋亡。结果番茄红素在2～10μmol·L-1浓度范围内的对肝细胞无毒性,可抑制CCl4所致肝细胞活性下降,减少CCl4所致的肝细胞凋亡,一定剂量范围内与番茄红素浓度相关。结论番茄红素对正常肝细胞HL-7702具有保护作用。

AN INTERMOLECULAR ^(13)C{~1H} NUCLEAR OVERHAUSER EFFECT FOR THE CARBON OF CCl_4 IN POLYMER SOLUTIONS

An intermolecular ^(13)C{~1H} NOE of CCl_4 in the solutions of polystyrene and polybutadiene and their copolymers was observed. The results show that the defined polymer-CCl_4 interaction variable has a linear relation with the polymer composition and the difference of solubility parameters and exponentially depends on the reciprocal of temperature.

绿豆癀对四氯化碳所致小鼠急性肝损伤的保护作用

目的:研究绿豆癀对四氯化碳(CCl_4)所致小鼠急性肝损伤的保护作用。方法:用CCl_4制作小鼠急性肝损伤模型,绿豆癀和绿豆粉干预,测定血清AST及ALT含量,并作肝组织病理学检测。结果:绿豆癀和绿豆粉能降低小鼠血清AST和ALT的含量(P<0.01或P<0.05);减轻肝细胞变性、坏死程度,缓解肝组织的病理改变,绿豆癀比绿豆粉效果好(P<0.05)。结论:绿豆癀对CCl_4所致小鼠急性肝损伤有保护作用。