Armored grass dimethylnitrosamine-induced inhibition of rat liver fibrosis

1、 Armored grass dimethylnitrosamine-induced inhibition of rat liver fibrosis Of: Zhang Ji Da, Wen Chengping, Fan Yongsheng Abstract Objective To observe the inhibition of ethanol extract of grass armored dimethylnitrosamine-induced liver fibrosis effect and its mechanism. Methods dimethylnitrosamine-

2、induced liver fibrosis in rats, with high, medium and low dose (12.6,4.2,1.4 g / kg) alcohol extract of grass armored intervention treatment, and were compared with colchicine, liver function test, serum hyaluronic acid (HA), layer mucin (LN and procollagen amino terminal peptide (PC ), liver hydrox

3、yproline (Hyp) content and liver histopathological changes. Results armored grass high, medium and low doses of serum alanine aminotransferase (ALT) and high-dose group armor grass serum aspartate aminotransferase (AST) levels compared with the model group was statistically significant (P <0.05),

4、 armored grass of liver tissue Hyp dose group were significantly lower than model group (P <0.05). Drafting of the doses of armored serum HA, LN, PC were significantly lower than the model group, the difference was statistically significant (P <0.05). armored grass high, medium and low dose gr

5、oup significantly reduced liver fibrosis, and model group, the difference was statistically significant (P <0.05). Conclusion armored grass on dimethylnitrosamine-induced liver fibrosis in rats significantly inhibited, and its mechanism may protect the liver cells and inhibition of collagen synth

6、esis related. Keywords: armored grass, hepatic fibrosis, dimethylnitrosamine Abstract: ObjectiveThe aim of this study was to observe the inhibitive effects of the extract of Cassia mimosoides Linn. On dimethylnitrosamine-induced hepatic fibrosis in rats and to explore the action mechanism. MethodsIn

7、tervention treatment with the extract of Cassia mimosoides Linn. Was explored in the rats with dimethylnitrosamine-induced hepatic fibrosis at the high, intermediate, and low doses of 12.6 g / kg, 4.2 g / kg, and 1.4 g / kg respectively, compared with the group treated with colchicine. Hepatic funct

8、ion, levels of serum hyaluronic acid (HA), serum laminin (LN), serum aminoterminal propeptide of type III procollagen (PIIINP), hepatic hydroxyproline (Hyp) in rats were assayed and histopathological alterations in rats were examined. ResultsThe results indicated that, compared with those in the mod

9、el group , alterations in the levels of serum alanine aminotransferase (ALT) in the groups treated with extract of Cassia mimosoides at low, intermediate, and high doses, and serum aspartate aminotransferase (AST) level in the group at the high dose were statistically significant (P <0.05); hepat

10、ic hydroxyproline (Hyp) levels in the rats treated with extract of Cassia mimosoides at various doses were significantly lower than those in the model group (P <0.05). Levels of serum hyaluronic acid (HA), serum laminin (LN ), and serum aminoterminal propeptide of type III procollagen (PIIINP) in

11、 the groups treated with extract of Cassia mimosoides were significantly lower than those in the model group, and the differences were statistically significant (P <0.05). Hepatic fibrosis of rats in the groups treated with extract of Cassia mimosoides at high, intermediate, and low doses allevia

12、ted significantly, and the differences compared with the pathological changes in the model group were statistically significant. ConclusionIt was concluded that, intragastric administration with extract of Cassia mimosoides showed significant inhibitive effects on the dimethylnitrosamine-induced hep

13、atic fibrosis in rats, and the inhibitive mechanisms might be related with the protective effects on hepatic cells and the inhibition on collagen fiber synthesis. Keywords: Cassia mimosoides Linn. (Leguminosae); Hepatic fibrosis; Dimethylnitrosamine Armored grass Cassia mimosoides Linn. As legume (L

14、eguminosae Cassia species, sweet, bitter, cool, the liver, kidney and lung, with Liver eyesight, and Water consumption carbuncle Tonglin and detoxification functions, can be used heat treatment of jaundice, dizziness, edema and carbuncle swollen boils, etc. 1. literatures armored grass contains emod

15、in, luteolin -7- grape glycoside, emodin, rhein 2,3, 1 ,8-dihydroxy-6-methoxy-2-methyl-anthraquinone 4. armored Meizhou in Guangdong Folk grass is mainly used for chronic hepatitis B and liver fibrosis, of armor for the grass on experimental liver fibrosis inhibition, we SD rats as experimental subj

16、ects, on dimethylnitrosamine-induced liver fibrosis model intervention to observe the effect of drug and its mechanism. 1 Materials and equipment 1.1 Animals SPF SD male rats, 60 rats weighing 160 180g, by the Guangdong Medical Experimental Animal Center, Permit No.: 2007A003, in Guangzhou Universit

17、y of Traditional Chinese Medicine Laboratory of Experimental Animal Center of SPF-level farming, animal testing facilities prove: 0022793 . rat chow by the Guangdong Medical Experimental Animal Center. Grass on the whole armor of 1.2 drugs purchased in the grass 2007-10 Jiaoling County, Meizhou, Gua

18、ngdong Province, with 85% ethanol extract 3 times, mixed with raw medicinal herbs concentrated into 1.26 g / ml, with distilled water when paired with the required concentration. Colchicine: Xishuangbanna Xishuangbanna Pharmaceutical Co., Ltd., Spec: 0.5 mg / tablets, batch number: 20071108, when us

19、ed with saline solution. 1.3 Reagents dimethylnitrosamine (DMN): purchased from Tianjin Chemical Reagent Research Institute, batch number: 060523, dubbed before use 0.5% saline solution with spare, aspartate aminotransferase (AST, alanine aminotransferase (ALT, triglyceride (TG, total cholesterol (C

20、HOL kit, the North Central Health Biotech Co., Ltd. production control, hyaluronic acid (HA), laminin (LN and N-terminal procollagen type peptide (PC ) kit, from the Beijing Institute of Biotechnology in North production, batch number: 080220, total protein (TP and albumin (AlB) test reagents from t

21、he distribution. 1.4 AU5421 automatic biochemical detector instrument (Olympus), FJ-2008PS Automatic immune counter (Xian Nuclear Instrument Factory), KDC02042 freezing centrifuge (HKUST Innovation Co., Ltd. in the good offices), TU -1001 UV-visible spectrophotometer (Shanghai Instrument Co., Ltd. c

22、losely), OlympusCK40 a F200 inverted microscope (Japan Olympusoptical company. 2 Methods 2.1 Animal grouping and drug modeling, 60 SD male rats were randomly divided into 6 groups of 10 each, respectively, the normal group, model group, armored grass extract high alcohol dose group, middle dose grou

23、p and low dose group ( 12.6,4.2,1.4 g / kg), colchicine treatment group (0.2 mg / kg). In addition to the normal group, the other animals in each group were prepared for liver fibrosis 5 to 0.5% DMN solution (1: 199 diluted with normal saline) according to the weight 10 mg / kg intraperitoneal injec

24、tion once a week consecutive 3 d, 1 times / d, 1 week each with 2 / 3 doses, after the whole amount with a total of 4 weeks, with an equal volume of normal of saline. in the preparation of models at the same time, armored grass high, medium and low-dose group and the colchicine treatment group were

25、gavage, 1 / d, 4 weeks, the normal group and model group herein are the physiological equivalent saline gavage. the first 4 weeks animals were killed eyeball blood serum was separated, to be seized, the left lobe of the liver tissue, part of the inspection to be made into homogenate, some were fixed

26、 with 10% formaldehyde, censorship. 2.2 liver function and liver fibrosis detected by automatic biochemical analyzer serum liver function, serum radioimmunoassay liver fibrosis. 2.3 Determination of liver tissue Hyp liver tissue were detected by alkaline hydrolysis Hyp, according to kit instructions

27、. 2.4 10% Histopathological observation of liver tissue formalin fixed, paraffin embedded sections, routine HE staining, light microscope photographs. By severe liver fibrosis were divided into five (S0 4) 6: S0 : no fibrosis, Sl: portal area, around the portal area fibrosis, limited sinus weeks or

28、lobular fibrosis, lobular structure does not affect the integrity of, S2: portal area around the fibrosis formation of fibrous septa, but a large lobular structure Department retained, S3: a large number of fibrous septa, and the destruction of hepatic lobules separated, lobular structural disorder

29、induced, but no liver cirrhosis, S4: early cirrhosis, extensive destruction of liver parenchyma, diffuse fibrosis, liver cell mass was separated was different levels of regeneration and the formation of false lobules. 2.5 Statistical Methods SPSS11.5 package for statistical analysis, measurement dat

30、a were analyzed using multiple One way ANOVA, multiple comparisons using LSD. Ridit level test using grouped data. Links to free paper download 3 Results 3.1 all the experimental rats after death, no rats died the normal group, model group, the colchicine group and the alcohol extract low dose group

31、 died 2 in each group, alcohol extraction, high-dose group died one. 3.2 Armored grass on liver function in rats with hepatic fibrosis affect the results in Table 1. Table 1 armored Grass on liver fibrosis serum ALT and AST of (slightly Table 1 shows that the modeling and After 4 weeks, model group

32、and other treatment group serum ALT, AST activity was significantly increased compared with normal control group and the difference was statistically significant (P <0.01 . Compared with model group, the dose of alcohol extract armored grass rats decreased serum ALT, with the model group was sign

33、ificantly different (P <0.05 or P <0.01, and only the high dose of alcohol extraction of serum AST and model group difference was statistically significant (P <0.05. With the colchicine group, armored grass alcohol extract of high dose of serum AST and colchicine group difference was statis

34、tically significant (P <0.05, suggest armor grass alcohol extract high dose dimethylnitrosamine AST increased significantly due to the reduced role, and is superior to colchicine group. As can be seen from Table 2, the modeling and After 4 weeks, each group serum TP, ALB and GLO significantly dec

35、reased, and the difference compared with the normal group was statistically significant (P <0.01, suggest liver fibrosis mode is ideal. Table 2 armored Grass on liver fibrosis serum TP, ALB and GLO effects (abbreviated Armored grass alcohol extraction and high dose of serum TP and ALB levels with

36、 colchicine group equivalent and compared with model group difference was statistically significant (P <0.05, while giving tips on armor modeling grass, grass alcohol armor extraction and high dose of dimethylnitrosamine-induced rat liver has a protective effect. 3.3 Grass on liver fibrosis in ra

37、ts armored CHOL, TG and liver Hyp content results in Table 3. Table 3 armored Grass on liver fibrosis in rats CHOL, TG and Hyp of (slightly As can be seen from Table 3, modeling and After 4 weeks, each group of CHOL and Hyp in serum was significantly increased. Alcohol extract of high dose of serum

38、CHOL in the normal group the difference was statistically significant ( P <0.05, the other group and no significant difference between the normal group. The serum TG were lower than in the normal group (P <0.01 or P <0.05. Use of dimethylnitrosamine after the model, the liver tissue of rats

39、 in each group Hyp increased significantly compared with the normal difference was statistically significant (P <0.01, while giving the grass treatment armor, armored grass alcohol extract of the large dose group Hyp mouse liver tissue was significantly lower than model group and model group diff

40、erence was statistically significant (P <0.05, prompt dose of ethanol extract of the liver fibrosis in rats have a certain effect, and its role and colchicine quite. 3.4 Grass on liver fibrosis in rats armored HA, LN and PC of results in Table 4. As can be seen from Table 4, the modeling and Afte

41、r 4 weeks, the serum HA, LN and PC increased significantly, compared with the normal group there was significant difference (P <0.01 or P <0.05. Table 4 Armored grass on serum hepatic fibrosis HA, LN and PC of (slightly Dose of ethanol extract of the grass armored rats serum HA, LN and PC decr

42、eased significantly, compared with model group difference was statistically significant (P <0.05, shows the dose of alcohol extracted HA, LN significantly elevated inhibition, and with colchicine quite. 3.5 Histopathological observation of liver tissue HE staining showed: normal rat liver cells a

43、rranged in neat bundles, was single, arranged radially around the central vein, no liver cell degeneration, necrosis and fibrosis changes, no expansion of sinusoidal hemorrhage model group of large Rat hepatic cord disorder, vacuolar degeneration of liver cells, showing large areas of hemorrhagic ne

44、crosis of liver cells in portal areas a large number of connective tissue proliferation, with marked hyperplasia of collagen fibers, a large fibrous septa, intralobular extension to form a pseudo lobular Most structural damage to rat hepatic lobule, a small amount of inflammatory cell infiltration,

45、armored grass alcohol extract high, medium and low-dose group compared with the model of liver cell degeneration and necrosis were significantly reduced degree of proliferation of connective tissue within the liver tissue was significantly reduced. Statistics The results showed that the grass high a

46、rmor, medium and low dose group significantly reduced liver fibrosis, compared with model group, the difference was statistically significant (P <0.05). The results in Table 5. Table 5 armored grass on rat liver fibrosis (slightly 4 Discussion Dimethylnitrosamine-induced by intraperitoneal inject

47、ion 3 d consecutive week, a total of 3 to 4 weeks is chemical poisoning induced liver fibrosis in the classical modeling approach, with model liver tissue was fibrous septa, and in exposed to stop characteristics not easily dissipated in the study of liver fibrosis is widely used 7. The results indicated that DMN-induced liver fibrosis in rats with chronic liver damage and significant liver fibrosis. the use of armored