| Background:Cirrhosis of the liver (hepatic sclerosis) is a clinical common chronic progressive liver disease, by one or more of the cause of "formed by the long-term or repeated diffuse liver damage. Have a wide range of pathologic histology liver cell necrosis, remnants of nodular liver cell regeneration, connective tissue hyperplasia and fiber forming, lead to hepatic lobule structure destruction and false flocculus to form, the liver deformation, hardening and for the development of liver cirrhosis. Clinical liver function damage and portal hypertension as the main performance, and how the system involved, often appear in late upper gastrointestinal bleeding, hepatic encephalopathy, and complications such as secondary infection. Main pathogenic mechanism of the present study suggests that liver cirrhosis has liver into fiber cell nucleus star cell proliferation and migration, inflammation caused by oxidative stress, and liver cell apoptosis. Rising in recent years, the incidence of liver cirrhosis, and because of its etiology pathology is complex, its pathogenesis and therapy intervention method is still unclear, has become the focus of social health problems.Carthamus tinctorius L. extract, including safflower yellow pigment and safflower glycosides, safflower glycosides by hydrochloric acid hydrolysis, glucose and red flowers; Also contains 15 alpha, beta,20-4-2 hydroxy-? pregnene-3-ketone; The other is fatty oil, safflower oil are palmitic acid, stearic acid, arachidic acid, oleic acid, linoleic acid and linolenic acid glyceride classes. Carthamus tinctorius L. extract, temperature, reduction and by the liver, anticoagulant, resisting thrombosis, increase coronary blood flow and reduce coronary artery resistance, dilate blood vessels, improve microcirculation, lowering blood pressure and blood fat, anti-inflammatory, analgesic, antioxidant, protecting liver, immune regulation, anti-tumor effect, etc. Experts often in the form of combination of traditional Chinese medicine will be red for the treatment of chronic liver disease or cirrhosis of the liver, but its inhibition mechanism of cirrhosis of the unknown. This topic with diethyl nitrosamine induced liver cirrhosis in rats model, to verify the safflower extract inhibition of cirrhosis of the liver, and the mechanism of its action will be discussed.Objective:Through the establishment of diethyl nitrosamine induced liver cirrhosis in rats model, discuss the inhibitory effect of safflower extract of cirrhosis of the liver and its mechanism of action. Providing theoretical basis for clinical application of safflower extract treatment cirrhosis of the liver.Method:1. Diethyl nitrosamine induced liver cirrhosis in rats model:30 healthy rats were randomly divided into three groups, each group of 10. The first group of normal group, intraperitoneal injection of saline once a week after oral saline (SI); The second group of diethyl nitrosamine induced rat liver cirrhosis model (control group), a weekly dose of 70. mg/2 mL/kg concentration diethyl nitrosamine (DEN), and then oral saline, deal with 8 weeks; A third group of diethyl nitrosamine induced liver cirrhosis in rats model (experimental group), with a weekly dose of 70 mg/2 mL/kg concentration diethyl nitrosamine (DEN), then the oral 40 mg/10 mL/kg (CTLE), once every day, concentration of safflower extract processing 8 weeks. Respectively in the processing of week 0, week 4 and 8 weeks under the orbital sinus in blood, and executed in 8 weeks rats in rat liver tissue, mitochondria, and other follow-up testing and comparison and analysis.2. each group of rat liver tissue, the formaldehyde fixed, paraffin embedding, frozen section biopsy, with HE staining to observe histomorphology structure change.3. spectrophotometry to detect plasma alanine aminotransferase, alanine transaminase, ALT and aspartate aminotransferase, aspartate aminotransferase, AST content.4. using ELISA method, plasma tumor necrosis factor TNF alpha, the concentration of TGF transforming growth factor beta.5. differential centrifugation separation of liver tissue cell mitochondria and cytoplasm.6. and transmission electron microscope hepatocyte mitochondrial morphology change.7. using colorimetry to detect malondialdehyde MDA and SOD of liver tissue and mitochondria.8. using spectrophotometry to detect the change of the mitochondrial membrane potential and mitochondrial ATP, Clark oxygen electrode method to detect mitochondrial respiratory function, observe mitochondrial damage.9. liver biopsy for TUNEL (TdT mediated dUTP Nick End Labeling) dyeing, observation of cell apoptosis.10. and Western Blot detecting cytochrome C in the organization, the protein content of Bax? Pro-caspase9 and caspase3.Results:1.safflower extracts to protect the liver tissue of cirrhotic model in rats(1) according to the results of tissue morphology of safflower extract processing group than the control group caused by rat liver cirrhosis of liver tissue injury and significantly reduce visible organization form relatively intact, necrotic tissue cell apoptosis, and inflammatory cells infiltrating significantly reduce, alleviate the swelling of liver cells, reduce liver tissue flbrosis, etc.(2) we found safflower extract could significantly lower in rat liver cirrhosis caused by plasma alanine aminotransferase, alanine transaminase, ALT and aspartate aminotransferase, aspartate aminotransferase, AST increases.(3) flower extract can significantly reduce liver cirrhosis in rats caused by tumor necrosis factor in plasma TNF alpha growth factors, the rise of the concentration of TGF beta.(4) Cirrhosis of the liver MDA level significantly increased in rat liver tissue, and the content of SOD decreased obviously, that liver cirrhosis increased oxidative stress in rat liver tissue; Safflower extract treatment can obviously reduce the levels of MDA, and increase the content of SOD. That safflower extract can effectively inhibit rat liver cirrhosis of liver tissue caused by oxidative stress.2. safflower extract effects on liver cirrhosis rat liver mitochondria.(1) to liver mitochondria were observed by using transmission electron microscopy (sem), we found that the blank group rats liver mitochondria neatly, form normal; In the control group rats swollen mitochondria and irregularly, boundary is not clear, and the size is inconsistent; And experimental mitochondria swelling, structure is not clear, sizes, and so on and so forth have improved significantly.(2) liver cirrhosis in rats of mitochondrial membrane potential energy greatly reduced compared with the blank group, treatment group of mitochondrial membrane potential and safflower extract is higher than the control group. This shows Carthamus tinctorius L. extract processing can help maintain their mitochondria membrane potential energy, maintain mitochondrial osmotic balance and protect the mitochondria structure from injury.3. safflower extract effects on liver cirrhosis rat liver mitochondrial function.(1) The level of MDA in liver cirrhosis rat hepatocyte mitochondria increased significantly, and the content of SOD decreased obviously, of cirrhosis of the liver mitochondria oxidative stress in rat liver tissue cells significantly enhanced; Safflower extract treatment can obviously reduce the levels of MDA, and increase the content of SOD. That safflower extract can effectively inhibit rat liver cirrhosis of liver tissue cell mitochondria oxidative stress.(2) in the rat can increase due to cirrhosis of the liver cells in the mitochondrial ATP consumption, increasing the extent of the oxidation reaction of mitochondria, and the consumption of safflower extract group of rats in the mitochondrial ATP significantly lower than the control group. Is consistent with the, safflower extract group p/o ratio of mitochondria of rats significantly higher than the control group.4. safflower extract apoptosis of the liver tissue of liver cirrhosis in rats(1) TUNEL staining results showed that apoptosis cells less blank group, and compared with blank group, control group obviously increase the rat liver tissue apoptosis, and safflower extract processing of the experimental group compared with control group apoptosis decreased obviously.(2) in blank group, the expression of Bax mainly in the cytoplasm, and the expression of Bax in control group significantly enhanced in the mitochondria, expressed in the cytoplasm decreased significantly, the experimental group was reduced compared with the control group in the expression of Bax change. That liver cirrhosis in rats liver damage, Bax in cells from the mass to the mitochondria transfer, while the safflower extract effectively block the transfer of Bax. The expression of cytochrome C and Bax show the opposite trend.(3) the control group obviously higher than that of blank group, and after processing, safflower extract Pro-caspase9 and caspase3 expression in liver cells was significantly decreased.Conclusion:This paper USES DEN induced liver cirrhosis in rats model and safflower extract in the model, and the rat liver tissue injury, inflammatory reaction, oxidative stress, mitochondrial function, cell apoptosis, such as testing and analysis, get the following conclusion:1. safflower extract could significantly reduce the liver cirrhosis in rats induced by DEN cause liver damage.2. safflower extract can effectively inhibit liver cirrhosis in rats caused by oxidative stress and inflammatory reaction.3. safflower extract could obviously reduce the liver cells of rat liver cirrhosis induced mitochondrial oxidative stress.4. safflower extract can maintain hepatocyte mitochondrial ATP levels, breath control and phosphorus oxygen ratios to protect the function of mitochondria.5. safflower extract can inhibit rat liver cirrhosis caused by liver cell apoptosis.6. safflower extract affects cytochrome C in liver cells and the expression of apoptosis proteins Bax? Pro-caspase9 and caspase3.Taken together, our study shows that the Carthamus tinctorius L. extract can inhibit rat liver cirrhosis induced cells mitochondrial respiratory function is impaired, the REDOX reaction of normal makes mitochondria, enhance its antioxidant ability, have the effect of reducing oxidative stress, and thus reduce the mitochondrial oxidative damage, in addition, safflower extract can maintain stability and maintain osmotic pressure in the mitochondria of mitochondrial membrane potential energy in a relatively normal levels, inhibit Bax by cytoplasm into mitochondrial cytochrome C and into the cytoplasm, thus inhibiting the expression of apoptosis factor Pro-caspase9 and caspase3, inhibiting cell apoptosis, oxidative stress, the liver tissue in order to reduce increase the antioxidant ability of organization, and can be gathered through the inhibition of inflammatory cells in the liver tissue and inflammatory cytokines released, significantly reduce the degree of inflammation tissue and thus effectively protect liver cirrhosis in rats caused by liver damage. For application of safflower extract in adjuvant treatment cirrhosis of the liver of rats caused by damage to provide theoretical support. |
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