Chinese herb Radix Polygoni Multiflori as a therapeutic drug for liver cirrhosis in mice

Liver regeneration not only plays a functional role in directing the restoration of liver mass after resection or injury, but also may have participated in effective therapy of liver cirrhosis. Additionally, hepatocyte growth factor (HGF) appears to be a factor of great importance in liver regeneration and attenuated progression of experimental liver cirrhosis. The aim of this study is to use Radix Polygoni Multiflori (POMU) extract, a Chinese herb traditionally used for liver-protective therapy, as a reagent for the evaluation of its potential medicinal use in liver cirrhosis. We used in vitro coculture system to show that POMU could promote the expression of HGF by hepatic nonparenchymal cells, consequently the proliferation of primary liver cells and phagocytic activity of Kupffer cells using fluorescein-labeled Escherichia coli as the target, and inhibit the proliferation of stellate cells. Using dimethylnitrosamine-induced liver cirrhosis animal, POMU even at 20 mg/(kg day) dosage, was illustrated to reverse the pathogenic progression of the disease, decrease the hydroxyproline content and increases the expression of HGF messenger RNA in liver tissue. The survival rate was significantly increased in the POMU-treated animal. In conclusion, our study showed the promise of POMU in the medicinal use for the treatment of liver cirrhosis.     

杠板归抗二甲基亚硝胺诱导大鼠肝纤维化的作用及其机制研究

目的:研究杠板归对二甲基亚硝胺(DMN)诱导的大鼠肝纤维化的作用及其机制.方法:80只SD大鼠随机分为正常组、模型组、杠板归高、中、低剂量组和秋水仙碱组共6组.除正常对照组以外,其余各组大鼠均以0.5％ DMN溶液(1.6mL·kg-1)腹腔注射,每周连续3天,每天1次,第1周用2/3剂量,以后用全量.秋水仙碱组剂量为0.1 mg·kg-1,杠板归高、中、低剂量组分别为7.5,5.0,2.5 g·kg-1,各组给药1次/d,连续4周.4周末摘大鼠眼球取血,酶联免疫吸附法(ELISA)检测血清透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C);苏木精-伊红染色(HE染色)观察肝纤维化(HF)的形成;免疫组化检测转化生长因子β1(TGF-β1).结果:与模型组比较,杠板归高、中剂量组大鼠血清HA,LN,PCⅢ,Ⅳ-C水平均显著低于模型组(P ＜0.05或P＜0.01),免疫组织化学显示杠板归高、中剂量组中大鼠肝脏TGF-β1表达明显降低(P ＜0.05或P＜0.01),低剂量无统计学意义(P＞0.05).结论:杠板归对二甲基亚硝胺诱导的大鼠肝纤维化具有较好的作用,作用机制可能与杠板归通过保肝、降低TGF-β1表达,从而抑制肝纤维化的启动.     

Induction of 6-thioguanine resistance in Chinese hamster lung cells treated with dimethylnitrosamine' 2-amino-anthracene or 7,12-dimethylbenz(A)anthracene in the presence of rat liver microsomes

Chemically non-reactive carcinogens require metabolization in order to exert such biological effects as mutagenicity. The mutagenic activities of dimethylnitrosamine (DMN), 2-aminoanthracene (2-AA) and 7,12-dimethylbenz(a)anthracene (7,12-DMBA) have been studied at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus in Chinese hamster lung cells. These compounds induce 6-thioguanine (6TG) resistance in the presence of rat liver microsomes, but they do not induce 6TG resistance in the absence of rat microsomes. The frequencies of mutations induced by these compounds are dose-dependent.     

冬虫夏草菌丝对二甲基亚硝胺诱导的大鼠肝纤维化的作用

目的:探讨冬虫夏草菌丝对二甲基亚硝胺诱导的大鼠肝纤维化的作用。方法:将SD大鼠分为正常对照组、模型组、冬虫夏草菌丝治疗组。采用二甲基亚硝胺诱导的大鼠肝纤维化模型。每组大鼠给予相应的药物。4周后,大鼠肝脏用胶原染色观察胶原纤维沉积的变化,盐酸水解法检测肝组织羟脯氨酸含量,Envi-sion两步法测定肝组织Ⅳ型胶原含量和金属蛋白酶组织抑制因子2(tissueinhibitorofmetalloproteinase-2,TIMP-2)含量;酶图法检测肝组织基质金属蛋白酶2(matrixmetalloproteinases-2,MMP-2)活性,Western印迹法检测肝组织Ⅰ型胶原。结果:模型组大鼠肝组织羟脯氨酸、Ⅳ型胶原和TIMP-2含量,I型胶原蛋白表达,较正常对照组均增加,而MMP-2含量降低;冬虫夏草菌丝治疗组大鼠肝组织羟脯氨酸、Ⅳ型胶原和TIMP-2含量,I型胶原蛋白表达较模型组均有不同程度的下降,而MMP-2含量升高。结论:冬虫夏草菌丝有显著的抗肝纤维化作用,其作用机制与促进胶原降解有关。     

Hepatic sinusoidal cell destruction in the development of intravascular coagulation in acute liver failure of rats

Rats received a dose of dimethylnitrosamine (DMN) or carbon tetrachloride (CCl4). In the liver of rats given DMN, apoptosis of fat-storing cells occurred at 7.5 h, and sinusoidal endothelial cell degeneration followed, with parenchymal cell necrosis after 9 h. Fibrin thrombi appeared in the sinusoids as well as in these necrotic areas after 12 h. In contrast, in the liver of rats given CCl4, parenchymal cell degeneration was seen after 6 h and necrosis with fibrin thrombi developed after 9 h. Fat-storing cells and endothelial cells were almost intact, and fibrin thrombi were not present in the sinusoids. SGPT values increased with decreased plasma levels of fibrinogen and antithrombin III and prolonged prothrombin time after 3 and 6 h, in the CCl4 and DMN models, respectively. An extensive reduction in plasma factor VIIIC levels and peripheral platelets was seen after 18 and 24 h, respectively, only in the DMN model. These results suggest that endothelial cells destruction can cause fibrin formation in the hepatic sinusoids in acute liver injury. Fat-storing cell injury may contribute to the destruction.     

罗格列酮对大鼠肝细胞色素P-4502E1活性的影响

目的 :探讨罗格列酮对细胞色素P 4 5 02E1活性的影响。方法 :在正常大鼠肝细胞微粒体中加入浓度为 0 ,0 .10 ,0 .2 5 ,0 .5 0 ,1.0 0mmol·L- 1罗格列酮 ,以N 二甲基亚硝胺为底物 ,测定细胞色素P 4 5 0 2E1活性。另外用 0 ,0 .2 ,2 ,2 0mg·kg- 1剂量的罗格列酮分别对用乙醇诱导和未用乙醇诱导的大鼠灌胃 ,然后取大鼠肝细胞微粒体 ,以N 二甲基亚硝胺为底物 ,测定细胞色素P 4 5 0 2E1活性。结果 :在体外试验中 ,加入不同浓度罗格列酮的大鼠肝微粒体中细胞色素P 4 5 0 2E1活性差异无显著意义。在体内试验中 ,给予不同剂量罗格列酮的大鼠的肝细胞色素P 4 5 0 2E1活性差异也无显著意义 ;结论 :罗格列酮对大鼠的肝细胞色素P 4 5 0 2E1活性无明显影响     

大蒜阻断亚硝胺的化学合成

大蒜提取液在pH3.15—3.35、37℃的条件下,能明显阻断二甲基亚硝胺、二乙基亚硝胺和二丁基亚硝胺的化学合成。其阻断机理在于大蒜中的有效成分能消除溶液中的NO_2~-。     

三甲益肝冲剂抗大鼠肝纤维化的实验研究

目的　观察三甲益肝冲剂 (SJYGG)对大鼠肝纤维化的防治作用。方法　 30只SD大鼠随机分为SJYGG预防组 (A组 )、模型组 (B组 )和SJYGG治疗组 (C组 )。各组均以二甲基亚硝胺 (DMN)腹腔注射 (每周连续 3d ,每天 1次 )共 4周 ,诱导肝纤维化模型。A组造模同时给予SJYGG(每日 6 .8g·kg-1)灌胃 ,共 8周。C和B两组造模 4周后 ,分别给予SJYGG(每日 6 .8g·kg-1)和生理盐水灌胃 ,共 4周。 8周后留取肝标本 ,行HE染色和Masson染色。结果　 (1)A组 :肝索排列基本整齐 ,有少量出血 ,汇管区稍扩大 ,未见胶原纤维增生。B组 :肝组织大片出血坏死 ,有假小叶形成。C组 :肝索排列紊乱 ,有出血坏死 ,汇管区扩大 ,有纤维间隔伸向肝小叶。 (2 )Masson染色胶原定量分析 :B组明显高于A、C两组 (P <0 .0 1) ,A组低于C组 (P <0 .0 5 )。结论　在DMN诱导的肝纤维化模型中 ,SJYGG具有预防和治疗肝纤维化的作用 ,且预防效果优于治疗。     

Suppression of macrophage infiltration inhibits activation of hepatic stellate cells and liver fibrogenesis in rats

BACKGROUND & AIMS: Monocytes/macrophages infiltrate into injured livers. We tried to clarify their roles in inflammation and subsequent fibrogenesis by inhibiting their infiltration with a mutated form (7ND; 7 amino acids at the N-terminal were deleted) of monocyte chemoattractant protein 1, which may function as a dominant-negative mutant. METHODS: Rats were injected via the tail vein with an adenovirus expressing either human 7ND (Ad7ND), a truncated type II transforming growth factor beta receptor (AdTbeta-TR), which works as a dominant-negative receptor, bacterial beta-galactosidase (AdLacZ), or saline. Seven days later, the rats were treated with dimethylnitrosamine for 1-21 days. RESULTS: Within 24 hours after a single dimethylnitrosamine injection, macrophages were observed in livers. With a 3-day dimethylnitrosamine treatment, activated hepatic stellate cells were detectable in livers in AdLacZ-, AdTbeta-TR-, and saline-injected rats. In contrast, in the Ad7ND-treated rats, infiltration of macrophages was markedly reduced, and activated hepatic stellate cells were not detectable. After a 3-week dimethylnitrosamine treatment, fibrogenesis was almost completely inhibited, and activated hepatic stellate cells were hardly seen in livers in both Ad7ND- and AdTbeta-TR-treated rats. CONCLUSIONS: Our results show that blockade of macrophage infiltration inhibits activation of hepatic stellate cells and leads to suppression of liver fibrogenesis. The presence of activated hepatic stellate cells in the initial phase after injury and its absence at a later phase in the AdTbeta-TR-treated livers indicate that transforming growth factor beta is not an activating factor for hepatic stellate cells, and this suggests that transforming growth factor beta is required for the survival of activated hepatic stellate cells. Our study suggests that infiltrated macrophages may themselves produce an activating factor for hepatic stellate cells.     

Decreased incidence of renal tumors in DMN treated Balb/C mice after orchiectomy

Dimethylnitrosamine (DMN) induced a significant incidence (45.5%) of kidney epithelial tumors in adult male Balb/C mice. The proportion of mice with renal tumors orchiectomized after DMN injection was reduced to 25.6%; whereas the proportion of tumor bearing animals at other sites, such as lung, liver, and lymphatic tissue, increased compared to intact DMN treated animals and controls. This experiment confirms other studies in rats and mice showing that androgens play a role in initiation and progression of nitrosamine induced renal cancer, paralleling the finding of increased male susceptibility in human renal cancer.