扶正化瘀方药物血清对原代培养纤维肝细胞胶原生成及分泌白蛋…

目的：研究扶正化瘀方对原代培养ＣＣｌ４纤维肝细胞胶原生成率及白蛋白（Ａ１ｂ）分泌功能的影响。方法：制备ＣＣｌ４肝纤维化模型，胶原酶原位灌流法分离纤维肝细胞，以＾３Ｈ－ＴｄＲ掺入，＾３Ｈ－Ｐｒｏｌｉｎｅ掺入胶原酶消化，ＥＬＩＳＡ等方法研究扶正化瘀方的血清药理学作用，结果：纤维肝细胞其增殖功能，Ａ１ｂ分泌功能的均较正常肝细胞低下，而胶原生成率则显著上升，扶正化瘀方药物血清能明显抑制纤维肝细胞增殖及其胶     

复方汉防己治疗实验性肝纤维化的机制探讨

目的：观察复方汉防己抗肝纤维化的疗效并探讨其机制。方法：用复方汉防己治疗ＣＣｌ４诱导的肝纤维化模型大鼠，观察它对血清透明质酸（ＨＡ），肝组织纤维化评分及储脂细胞的影响，ＨＡ测定采用放免法，储脂细胞观察采用免疫组化法。结果：复方汉防己治疗组肝纤维化评分明显低于生理盐水对照组，血清ＨＡ及结蛋白阳性细胞数明显减少。结论：复方汉防己有良好的抗肝纤维化作用，其机制可能是抑制储脂细胞的增殖。     

珠子肝泰胶囊对大鼠肝纤维化的作用

目的：观察珠子泰胶囊（ＺＧＴ）的抗肝纤维化作用。方法：用四氯化碳（ＣＣｌ４）诱导大鼠肝纤维化模型,用ＺＧＴ治疗,观察其对大鼠肝功能、超氧化物歧化酶（ＳＯＤ）、丙二醛（ＭＤＡ）、透明质酸（ＨＡ）、Ⅲ型前胶原（ＰＣⅢ）、层粘蛋白（ＬＮ）及肝脏组织病理学的影响;同时设正常对照组及阳性对照组。结果：ＺＧＴ能显著降低大鼠血清转氨酶、ＭＤＡ、ＨＡ、ＰＣⅢ和ＬＮ,提高肝组织中ＳＯＤ的活性;明显减轻肝脏纤维增生程度。结论：ＺＧＴ对大 实验性肝纤维化有预防作用。     

肝刺激因子对实验性免疫性肝纤维化保护作用

目的：研究肝刺激因子（ＨＳＳ）对免疫性肝纤维化的作用。方法：ＨＳＳ从初生小牛肝提取，动物模型采用人血清白蛋白（ＨＳＡ）制备，保护组同时腹腔注射８ｍｇＨＳＳ。结果：（１）ＨＳＳ可使升高的血清转氨酶（ＡＬＴ）水平、Ｎ－乙酶－β－Ｄ－氨基葡萄糖苷酶（ＮＡＧ）活力及肝组织羟脯氨酸（ＨＹＰ）含量下降。（２）光、电镜下见，ＨＳＳ可以保护肝细胞的受损，使成纤维细胞、Ｉ型及Ⅲ型胶原增生程度明显降低。（３）ＨＳＳ使贮脂细胞数目明显减少且影响其激活，并且减少了肝内ＨＳＡ免疫复合物的沉积及大量浸润的Ｔ细胞。结论：ＨＳＳ对免疫性肝纤维化有保护作用，其机制可能是通过免疫途径。     

褐藻胶对大鼠实验性肝纤维化的防治作用

目的研究褐藻胶对大鼠实验性肝纤维化的防治作用．方法用４０％四氯化碳（ＣＣｌ４）制备大鼠肝纤维化模型，实验分组为正常对照组（ｎ＝８），ＣＣｌ４组（ｎ＝８），秋水仙硷（ＣＯＬ）组（ｎ＝６）和褐藻胶组（ｎ＝６）（２００ｍｇ／ｋｇ，ｉｇ，３次／周×４）．观察肝脏组织学和血清Ⅲ型前胶原（ＰＣⅢ）及透明质酸（ＨＡ）水平的变化．结果褐藻胶组ＰＣⅢ（１４２８μｇ／Ｌ±７６１μｇ／Ｌ）和ＨＡ水平（２６５５μｇ／Ｌ±９３１μｇ／Ｌ）显著低于ＣＣｌ４组（２９３５μｇ／Ｌ±７８３μｇ／Ｌ，５１９８μｇ／Ｌ±１１８３μｇ／Ｌ，Ｐ＜００１），而褐藻胶组与ＣＯＬ组间无显著差异．病理学观察显示，ＣＣｌ４组肝纤维化程度重于褐藻胶组和ＣＯＬ组．结论褐藻胶对实验性肝纤维化有防治作用．     

牛磺酸对四氯化碳诱导大鼠肝纤维化的抑制作用

目的研究牛磺酸的体内抗肝纤维化作用。方法用四氯化碳（ＣＣｌ４）复制大鼠肝纤维化模型;造模开始或造模６周后给予牛磺酸;实验结束后分别测定肝功能、纤维化指标（透明质酸和层粘连素）、肝羟脯氨酸及Ⅰ、Ⅲ型前胶原ｍＲＮＡ含量,并作肝病理检查。结果牛磺酸可显著减轻ＣＣｌ４肝纤维化程度,能明显降低肝羟脯氨酸和Ⅰ、Ⅲ型前胶原ｍＲＮＡ含量,降低血清透明质酸和层粘连素水平,改善肝功能,组织学检查亦显示具有抗肝纤维化作用。结论牛磺酸在体内具有抗肝纤维化的作用,可望用于肝纤维化的防治。     

虫草多糖对大鼠Ito细胞增殖及胶原基因表达的影响

目的　研究虫草多糖（ＣＰ）对大鼠Ｉｔｏ细胞增殖及Ⅰ、Ⅲ型前胶原基因ｍＲＮＡ表达的影响,探索ＣＰ抗肝纤维化的作用机制。方法分离、培养大鼠Ｉｔｏ细胞,采用~３Ｈ－胸腺嘧啶（~３Ｈ－ＴｄＲ）和~３Ｈ－脯氨酸（~３Ｈ－Ｐｒｏ）掺入法测定Ｉｔｏ细胞增殖及胶原合成;用原位杂交法检测Ｉｔｏ细胞Ⅰ、Ⅲ前胶原基因ｍＲＮＡ含量。结果　在０．１～８０μｇ／ｍ剂量范围内,ＣＰ可明显抑制Ｉｔｏ细胞~３Ｈ－ＴｄＲ和~３Ｈ－Ｐｒｏ的掺入。当ＣＰ浓度为１０μｇ／ｍｌ时,抑制率分别为２７．９％和４５．４％（Ｐ均＜０．０５）。随着浓度的增加和作用时间的延长,其抑制作用逐渐加强,１０μｇ／ｍｌ ＣＰ可显著抑制Ⅰ、Ⅲ型前胶原ｍＲＮＡ的表达,抑制率分别为３２．６０％和３７．３２％（Ｐ均＜０．０１）。结论 ＣＰ在体外可抑制Ｉｔｏ细胞的增殖和胶原合成,下调Ⅰ、Ⅲ型前胶原ｍＲＮＡ表达,并呈剂量和时间依赖性,提示ＣＰ对Ｉｔｏ细胞增殖和胶原合成的抑制可能是其体内抗肝纤维化作用的主要途径。     

血府逐瘀汤分解方抗肝纤维化作用的研究

以ＳＤ大白鼠为实验动物，用ＣＣｌ４诱导肝硬化模型，以观察血府逐瘀汤分解方的作用。提示桃红四物汤具有极好的抗肝纤维化作用，桃红四物汤Ⅰ、Ⅲ型胶原均具有明显抑制作用，且其抑制Ⅰ型胶原作用优于抑制Ⅲ型胶原。     

牛磺酸抑制实验性肝纤维化大鼠细胞外基质沉积

目的研究牛磺酸抗肝纤维化作用。大法用四氯化碳（ＣＣＩ４）制备肝纤维化模型;免疫组化检测肝Ⅰ、Ⅲ、Ⅵ型胶原和透明质酸、层粘连素沉积;Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交检测肝Ⅰ、Ⅲ型前胶原、组织金属蛋白酶抑制因子－１（ＴＩＭＰ—１）ｍＲＮＡ含量。结果ＣＣＩ４肝纤维化大鼠肝脏Ⅰ、Ⅲ、Ⅵ型胶原、透明质酸和层粘连素染色明显增多;Ⅰ、Ⅲ型前胶原、ＴＩＭＰ－１ｍＲＮＡ显著增加。牛磺酸处理的大鼠,上述各指标的阳性染色明显减少,并可明显抑制Ⅰ、Ⅲ型前胶原基因表达,而对ＴＩＭＰ—１ｍＲＮＡ却无明显影响。结论牛磺酸对ＣＣＩ４诱导的大鼠肝纤维化具有保护作用,可明显抑制纤维生成与沉积。提示它对防治肝纤维化有一定的临床意义。     

大黄Zhe虫丸抗慢性肝损伤及肝纤维化作用的实验研究

本文以ＣＣｌ４等因素所致大鼠肝损伤、肝纤维化为模型，以血清ＡＬＴ活民生、Ａｌｂ含量、肝Ｈｙｐ含量、肝组织ＨＥ和胶原染色光镜观察等为观察内容，分预防和治疗实验观察大黄Ｚｈｅ虫丸的防治作用，并设西药秋水仙碱组对照。结果表明：大黄Ｚｈｅ虫丸有较显著的抗慢性肝损伤作用及一定的抗肝纤维化作用。在抗肝损伤，保护肝功能产秋水仙碱为优；但在抗肝纤维化方面，高不及秋水仙碱作用强。     

血清羟脯氨酸和透明质酸对当归补血汤治疗兔血吸虫病肝纤维化的效果评价

 目的探讨血清羟脯氨酸(HYP)、透明质酸(HA)在当归补血汤治疗血吸虫病肝纤维化中的疗效考核作用。方法用日本血吸虫尾蚴感染日本大耳白兔，建立血吸虫病肝纤维化模型，随机将感染兔分成2个实验组(A、B组)和对照组(C组)，分别于服药前及服药10周后检测HYP及HA含量，同时解剖兔模做肝组织病理学检查。结果实验组服药10周后血清HYP、HA含量较服药前明显降低(P<0.01)，对照组则无明显变化(P>0.05)；组织病理学肝纤维化程度与血清HYP、HA含量呈正相关关系。结论当归补血汤对兔血吸虫病肝纤维化有治疗作用；血清HYP、HA是肝纤维化疗效考核较好的无创性检测指标。      

Melatonin ameliorates experimental hepatic fibrosis induced by carbon tetrachloride in rats

AIM： To investigate the protective effects of melatonin on carbon tetrachloride （CCl4）-induced hepatic fibrosis in experimental rats.
METHODS： All rats were randomly divided into normal control group, model control group treated with CCl4 for 12 wk, CCl4 ＋ NAC group treated with CCl4 ＋ NAC （100 mg/kg, i.p.） for 12 wk, CCl4 ＋ MEL-1 group treated with CCl4 ＋ melatonin （2.5 mg/kg） for 12 wk, CCl4 ＋ MEL-2 group treated with CCl4 ＋ melatonin （5.0 mg/kg） for 12 wk, and CCl4 ＋ MEL-3 group treated with CCl4 ＋ melatonin （10 mg/kg）. Rats in the treatment groups were injected subcutaneously with sterile CCl4 （3 mL/kg, body weight） in a ratio of 2：3 with olive oil twice a week. Rats in normal control group received hypodermic injection of olive oil at the same dose and frequency as those in treatment groups. At the end of experiment, rats in each group were anesthetized and sacrificed. Hematoxylin and eosin （HE） staining and Van Gieson staining were used to examine changes in liver pathology. Serum activities of alanine aminotransferase （ALT）, aspartate aminotransferase （AST） and protein concentration weremeasured with routine laboratory methods using an autoanalyzer. Hydroxyproline （HYP） content in liver and malondialdehyde （MDA） and glutathione peroxidase （GPx） levels in liver homogenates were assayed by spectrophotometry. Serum hyaluronic acid （HA）, laminin （LN）, and procollagen Ⅲ N-terminal peptide （PⅢNP） were determined by radioimmunoassay.
RESULTS： Pathologic grading showed that the fibrogenesis was much less severe in CCl4 ＋ MEL3 group than in model control group （u = 2.172, P 〈 0.05）, indicating that melatonin （10 mg/kg） can significantly ameliorate CCl4-induced hepatic fibrotic changes. The serum levels of ALT and AST were markedly lower in CCl4 ＋ MEL treatment groups （5, 10 mg/kg） than in model control group （ALT： 286.23 ± 121.91 U/L vs 201.15 ± 101.16 U/L and 178.67 ± 103.14 U/L, P = 0.028, P = 0.007; AST： 431.00 ± 166.35 U/L vs 321.23 ± 162.48 U/L and 292.42 ± 126.23 U/L, P = 0.043, P = 0.013）. Similarly, the serum laminin （LN） and hyaluronic acid （HA） levels and hydroxyproline （HYP） contents in liver were significantly lower in CCl4 ＋ MEL-3 group （10 mg/kg） than in model control group （LN： 45.89 ± 11.71 μg/L vs 55.26 ± 12.30 μg/L, P = 0.012; HA： 135.71±76.03 μg/L vs 201.10 ± 68.46 μg/L, P = 0.020; HYP： 0.42 ± 0.08 mg/g tissue vs 0.51 ± 0.07 mg/g tissue, P = 0.012）. Moreover, treatment with melatonin （5, 10 mg/kg） significantly reduced the MDA content and increased the GPx activity in liver homogenates compared with model control group （MDA： 7.89 ± 1.49 noml/mg prot vs 6.29 ±1.42 noml/mg prot and 6.25 ±2.27 noml/mg prot, respectively, P = 0.015, P = 0.015; GPx： 49.13 ±8.72 U/mg prot vs 57.38 ±7.65 U/mg prot and 61.39 ±13.15 U/mg prot, respectively, P = 0.035, P = 0.003）.
CONCLUSION： Melatonin can ameliorate CCl4 -induced hepatic fibrosis in rats. The protective effect of melatonin on hepatic fibrosis may be related to its antioxidant activities,     

肝泰胶囊抗肝纤维化作用的实验研究

目的 探讨肝泰胶囊对CCl4诱导的肝纤维化大鼠的作用及其抗肝纤维化的可能机制。方法将40只Wistar大鼠随机分为正常对照组、肝纤维化模型组、肝泰组及肝复乐组。采用光镜、电镜观察肝组织病理学改变，VonGieson（VG）染色及胶原面积半定量分析反映各组肝纤维化程度。采用放射免疫法测定血清层粘连蛋白（LN）、Ⅲ型前胶原（PC-Ⅲ）、Ⅳ型前胶原（Ⅳ-C）、透明质酸（HA）的含量，同时用高效液相色谱-电化学（HPLC—ECD）法检测肝组织中去甲肾上腺素（Norepinephrine，NE）的水平。结果肝泰组较模型组比：①肝组织肝纤维化程度明显改善；②肝组织胶原纤维面积明显减少；③血清透明质酸和层粘连蛋白显著降低；④肝功能损伤明显减轻；⑤组织中NE的含量明显高于模型组。结论肝泰对慢性肝损伤有一定的保护作用。     

体外四氯化碳损伤肝细胞对肝星状细胞活化的影响及其丹酚酸A的干预效果

目的 体外观察肝细胞四氯化碳过氧化损伤后肝星状细胞（ＨＳＣ）活化的影响,并观察丹参水溶性成分丹酚酸Ａ的干预效果,以研究脂质过氧化损伤与肝纤维化的关系和丹酚酸Ａ的作用机理。方法 肝细胞分离后分组,分别添加不同浓度的丹酚酸Ａ以及对照药维生素Ｅ,同时以四氯化碳熏蒸２４ｈ并测定各组细胞上清液ＡＬＴ、ＡＳＴ活性后,换液培养２４ｈ并测定ＭＤＡ含量,再以此上清液作为“肝细胞条件培养液”作用于ＨＳＣ４８ｈ,观察Ｈ     

虫草菌丝抗肝纤维化的实验研究

目的:观察人工虫草菌丝对CCl_4中毒性肝纤维化的预防作用。方法:复制大鼠CCl_4中毒性肝纤维化模型,并以秋水仙碱作为阳性治疗对照,采用光镜、电镜观察肝脏组织学改变,测定血清ALT、Alb、G、HA、LN、肝组织Hyp含量。结果:虫草菌丝组肝细胞损害、肝脏脂肪变性、炎性细胞浸润的程度均较模型组轻,但再生肝细胞较多,有假小叶形成;其对肝功能、肝纤维化指标也有明显改善,但秋水仙碱的抗肝纤维化作用优于虫草菌丝。结论:虫草菌丝对肝脏CCl_4损伤具有一定保护作用,有较强促进肝细胞再生修复作用,同时在损伤肝组织中成纤维细胞及胶原纤维也明显增生,其与活血化瘀药有机伍用,可能是防治肝纤维化较理想的组合。     

绯红南五味子对大鼠四氯化碳肝损伤的保护作用

目的 :探讨中药绯红南五味子对肝损伤的保护作用。方法 :采用四氯化碳加饲养高胆固醇制作大鼠肝损伤模型 ,同时给大鼠服用绯红南五味子水煎剂 ,连续服用 6周后 ,将动物处死 ,用分光光度法测定肝组织内丙二醛 (MDA)、超氧化物歧化酶 (SOD)、羟脯氨酸 (Hyp)的含量变化。结果 :与模型组比较 ,绯红南五味子 5 g/ kg能明显降低肝损伤时肝组织中 MDA和 Hyp的含量 (P <0 .0 5 ) ,提高 SOD活性。结论 :绯红南五味子能降低肝组织脂质过氧化 ,提高氧自由基清除能力 ,对肝损伤具有一定的保护作用。     

Therapeutic effects and molecular mechanisms of anti-fibrosis herbs and selenium on rats with hepatic fibrosis

AIM:To study the therapeutic effects of anti-fibrosis herbs and selenium on hepatic fibrosis induced by carbon tetrachloride (CCl4) in rats and the underlining molecular mechanisms.METHODS:Fifty-three Wistar rats were randomly divided into:normal control group, model control group, colchicine group, anti-fibrosis herbs group (AF group) and anti-fibrosis herbs plus selenium group (AS group).The last four groups were administered with CCl4 at the beginning of experiment to induce hepatic fibrosis. Then colchicine, anti-fibrosis herbs and selenium were used to treat them. The normal control group and the model control group were given normal saline at the same time. At the end of the 6^th week, rats in each group were sacrificed. Blood and tissue specimens were taken. Serum indicators (ALT, AST, HA, LN) were determined and histopathological changes were graded. Lymphocyte CD4 and CD8 were examined by flow cytometry. Expression of TGF-β1 and NF-κB was detected by immunohistochemistry and expression of TGF-β1 mRNA was detected by semiquantified RT-PCR.RESULTS: Histological grading showed much a smaller degree of hepatic fibrogenesis in AS group and AF group than that in colchicine group and model control group.The serum content of ALT, AST, HA and LN in AF group and AS group were significantly lower than that in colchicine group (ALT:65.8&#177;26.5, 67.3&#177;18.4 and 96.2&#177;20.9 in AF, AS and colchicine groups respectively; AST: 150.8&#177;34.0, 154.6&#177;27.3 and 215.8&#177;24.6 respectively; HA: 228&#177;83, 216&#177;58 and 416&#177;135 respectively; LN:85.9&#177;15.0, 80.6&#177;18.6 and 106.3&#177;14.2 respectively) (P&lt;0.05). The level of CD4 and CD4/CD8 ratio in AF group and AS group was significantly higher that those in cochicine group (CD4:50.8&#177;3.8, 52.6&#177;3.4 and 40.2&#177;2.1 in AF, AS and colchicine groups respectively;CD4/CD8 ratio:1.45,1.46 and 1.26, respectively (P&lt;0.05).The expression level of NF-κB and TGF-β1 in the liver tissues of AF and AS treatment groups was markedly decreased compared with that in cochicine group, and TGF-β1 mRNA was also markedly decreased (1.07&#177;0.31 and 0.98&#177;0.14 vs2.34&#177;0.43, P&lt;0.05).CONCLUSION: Anti-fibrosis herbs and selenium have beneficial effects on hepatic fibrosis in rats by enhancing immunity and inhibiting NF-κB and TGF-β1 expressions.     

Inhibitory effects of saikosaponin-d on CCl4-induced hepatic fibrogenesis in rats

AIM： To investigate the suppressive effect of saikosaponin-d （SSd） on hepatic fibrosis in rats induced by CCh injections in combination with alcohol and high fat, low protein feeding and its relationship with the expression of nuclear factor-κB （NF-κB）, tumor necrosis factor-alpha （TNF-α） and interleukins-6 （IL-6）. METHODS： Hepatic fibrosis models were induced by subcutaneous injection of CCh at a dosage of 3 mL/kg in rats. At the same time, rats in treatment groups were injected intraperitoneally with SSd at different doses （1.0, 1.5 and 2.0 mg/kg） once daily for 6 wk in combination with CCh, while the control group received olive oil instead of CCh. At the end of the experiment, rats were anesthetized and killed （except for 8 rats which died during the experiment; 2 from the model group, 3 in high-dose group, 1 in medium-dose group and 2 in lowdose group）. Hernatoxylin and eosin （HE） staining and Van Gieson staining were used to examine the changes in liver pathology. The levels of alanine aminotransferase （ALT）, triglyeride （TG）, albumin （ALB）, globulin （GLB）, hyaluronic acid （HA） and larninin （LN） in serum and the content of hydroxyproline （HYP） in liver were measured by biochemical examinations and radioimmuneoassay, respectively. In addition, the expression of TNF-α and IL-6 in liver homogenate was evaluated by enzymelinked immunosorbent assay （ELISA） and the levels of NF-κBp65 and I-κBa in liver tissue were analyzed by Western blotting. RESULTS： Both histological examination and Van Gieson staining demonstrated that SSd could attenuate the area and extent of necrosis and reduce the scores of liver fibrosis. Similarly, the levels of ALT, TG, GLB, HA, and LN in serum, and the contents of HYP, TNF-α and IL-6 in liver were all significantly increased in model group in comparison with those in control group. Whereas, the treatment with SScl markedly reduced all the above parameters compared with the model group, especially in the medium gr     

血吸虫病肝纤维化程度与血清透明质酸和肿瘤坏死因子水平的关系

本文报道血吸虫病肝纤维化兔与病人（晚期）的血清透明质酸（HA）和肿瘤坏死因子－α（TNF）与肝纤维化程度的关系。感染日本血吸虫新西兰兔随机分为两组，其中一组经杀虫治疗，均在感染后28周解剖，两组间肝纤维伦程度有明显差异（P＜0．001）。血清TNF水平与肝组织学检查肝纤维化程度，肝胶原含量及血清肝纤维化指标HA等相关。晚期病人亦获类似的结果。提示血清TNF水平可作为肝纤维化一个较好的指标。     

维生素E对肝纤维化大鼠肝脏病变及血型胶原表达的影响

为了解补充维生素E(VE)对肝纤维化的影响,以四氯化碳(CCl_4)诱导大鼠肝纤维化模型,对VE(100mg/kg,2次/周,连续10～20周)抗肝纤维化的作用进行了研究。于治疗后10周、20周取肝组织作病理检查及Ⅲ型胶原免疫组化染色,同时设溶剂对照组及盐水对照组。结果表明,经VE治疗后,肝纤维化大鼠肝脏胶原纤维及网状纤维由粗大变纤细并有缩短及断裂;免疫组化染色发现肝纤维化大鼠肝内Ⅲ型胶原主要位于肝细胞浆,呈弥漫性分布,VE治疗组含量较对照组减少,经真彩色图像系统分析与对照组差异显著(P<0.01)。提示VE治疗有助于实验性肝纤维化的恢复。