ABSTRACT
ObjectiveTo explore the inhibitory effect of water extract of Broussonetiae Fructus on hepatocellular carcinoma （HCC） induced by diethyl nitrosamine （DEN） in mice based on homologous phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B （PTEN/PI3K/Akt） signaling pathway. MethodThe primary HCC mouse model was constructed by intraperitoneal injection of DEN solution， and the HCC mice were randomly divided into model group， sorafenib group （0.01 g·kg-1·d-1）， low-dose Broussonetiae Fructus water extract group （0.9 g·kg-1·d-1）， medium-dose Broussonetiae Fructus water extract group （1.8 g·kg-1·d-1）， and high-dose Broussonetiae Fructus water extract group （3.6 g·kg-1·d-1）， with 10 mice in each group. Another 10 C57BL/6 mice were selected as a control group and intraperitoneally injected with an equal volume of normal saline. Mice were treated with different concentrations of Broussonetiae Fructus water extract when liver cancer-like white nodules appeared. sorafenib group was treated with sorafenib. The control group and model group were intraperitoneally injected with normal saline. The activities of alkaline phosphatase （ALP）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， and γ-glutamyl transferase （γ-GT） in the serum of mice were detected by the biochemical analyzer. The expression levels of alpha-fetoprotein （AFP） and carcinoembryonic antigen （CEA） were detected by enzyme-linked immunosorbent assay （ELISA）. The degree of hepatocyte canceration and hepatocyte injury were observed by Hematoxylin-eosin （HE） and Masson staining. The proliferation of HCC cells was observed by immunohistochemical staining. The apoptosis of HCC cells in mice was observed by erminal-deoxynucleotidyl transferase mediated nick end labelling （TUNEL） staining. The expression levels of PTEN， PI3K， Akt， and p-Akt proteins related to the PTEN/PI3K/Akt signaling pathway were detected by Western blot. ResultCompared with the control group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the model group were significantly increased （P<0.01）. Carcinogenesis and inflammatory cell infiltration were obvious in liver tissue of mice， and a large number of blue collagen fiber hyperplasia was found. The number of Ki67 positive cells was significantly increased （P<0.01）， and the expression level of PTEN protein was significantly decreased， while PI3K and p-Akt protein expression was increased （P<0.01）. Compared with the model group， the activities of ALP， AST， ALT， and γ-GT， as well as the expression levels of AFP and CEA in the medium-dose and high-dose Broussonetiae Fructus water extract groups were significantly decreased （P<0.05， P<0.01）. The degree of carcinogenesis and inflammatory cell infiltration in liver tissue were reduced， and the collagen fiber hyperplasia was significantly reduced. The number of Ki67 positive cells was significantly decreased， and the number of TUNEL positive apoptotic cells was significantly increased （P<0.05， P<0.01）. PTEN protein expression was increased， while p-Akt protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionThe water extract of Broussonetiae Fructus has a significant inhibitory effect on DEN-induced primary HCC in mice， and its mechanism may be related to the regulation of key protein expressions in the PTEN/PI3K/Akt signaling pathway.
ABSTRACT
Objective To investigate the protective effect of safflower extract on hepatocyte mitochondria in hepatic fibrosis rats.Methods Rat model of hepatic fibrosis was induced by diethyl nitrosamine.Oral safflower extract was taken orally in the experimental group.Hepatic mitochondria were isolated from each group.Morphological changes of hepatocyte mitochondria were observed and MDA and SOD were detected in the level hepatocyte mitochondria.Observe changes in mitochondrial membrane potential and mitochondrial ATP levels and observe changes in mitochondrial respiratory function to investigate whether safflower extract has protective effects on hepatocyte mitochondria in rats with hepatic fibrosis or not.Results In the normal group,the rat mitochondria were arranged neatly and the morphology was normal.In the control group,the mitochondria of the rats were swollen with irregular morphology,the borders were unclear,and the sizes were inconsistent.In the experimental group,the mitochondria were swollen,the structures were not clear,and the sizes were different.The above condition is significantly improved;liver fibrosis in rats' mitochondria MDA levels increased significantly,while SOD content decreased significantly,safflower extract treatment can significantly reduce mitochondrial MDA levels,and it can increase SOD content.Compared with the normal group,the mitochondrial membrane potential of liver fibrosis rats was significantly reduced,while the safflower extract treatment can increase the mitochondrial membrane potential;due to liver fibrosis,rats can increase the consumption of ATP in liver mitochondria and safflower extract can significantly reduce ATP consumption in rat mitochondria.Consistent with this,the phosphor-to-oxygen ratio in the rat mitochondria of the experimental group was significantly higher than that of the control group.Conclusion The safflower extract can significantly reduce the hepatocyte mitochondrial oxidative stress induced by hepatic fibrosis in rats.It can maintain mitochondrial ATP level,respiratory control rate and phosphorus-oxygen ratio to protect mitochondrial function.
ABSTRACT
Oxidative stress reflects the mechanism that contributes to initiation and progression of hepatic injury in a variety of liver disturbance. From here, there is a great demand for the expansion of agents with a potent antioxidant effect. The aim of this work is to approximate the efficiency of bee honey as a hepatoprotective and an antioxidant agent versus diethyl nitrosamine (DEN) motivate hepatocellular damage. The single intrapritoneal (IP) management of diethyl nitrosamine (50mg/kg followed by 2ml/kg CCl4) to rats, referred for the histopathological examination of liver sections of rats after induction and before treatment with honey showed that many well differentiated tumor cells were formed in the liver of rats also, the examined sections showed disorganization of hepatic lobular architecture and obvious cellular damage. A significant lift in the enzymatic activity of liver functions (AST, ALT, ALP), and gamma glutamyltransferase (GGT) which is a signal of hepatocellular damage. DEN stimulates oxidative stress, which was assured by increase lipid peroxidation level and hindrance in antioxidant enzymes (SOD, CAT, GPx, and GST) activities in the liver. The position of non-enzymatic antioxidants comparable reduced glutathione (GSH) was likewise set up to be slimmed down significantly in DEN inoculated rats. Also, we have studied the underlying mechanism and /or (s) of the therapeutic role of bee honey as hepatocarcinogenesis remediation through investigation the inflammatory biomarkers; α-fetoprotein (AFP) and α-fucosidase (AFU). The current results clearly showed that bee honey demonstrates good ameliorative and antioxidant capacity toward diethyl nitrosamine induced hepatocellular damage in rats.
ABSTRACT
Plant derived drugs have been a part of the evolution of human healthcare for thousands of years. The present study was carried out to evaluate the antioxidant potential of ethanolic extract of Tabernaemontana divaricata on DEN initiated and Fe-NTA promoted renal damage in rats. Fe-NTA was induced after 10 days of DEN (200mg/kg body weight) initiation at a dose level of 9mg Fe-NTA/kg body weight twice a week for one month. The biochemical parameters were analyzed in serum and the antioxidant assays were carried out in kidney. Lipid peroxidation level was increased due to the administration of Fe-NTA, which caused the reduction of enzymatic antioxidant such as SOD, GPx, Catalase, G6PD, and also the non-enzymatic antioxidants vitamin C and GSH. The levels of urea, uric acid, creatinine, blood urea nitrogen were increased and protein level decreased on Fe- NTA intoxication. The secondary metabolites present in the plant increased the synthesis of antioxidant enzymes and its free radical scavenging properties helped to scavenge all free radicals thereby decreasing lipid peroxidation. Thus, the present study indicates that the plant may clinically valuable agent in the prevention of renal failure caused by DEN and Fe-NTA intoxication.
ABSTRACT
Water gets magnetically charged when it is contacted with a magnet. Although magnetic water products have been promoted since the 1930's, they have received very little recognition due to questionable effectiveness. Diethylnitrosamine (DEN) is a widely occurring nitrosamine that is one of the most important environmental carcinogens primarily inducing tumors of liver. In this study, the effect of magnetized water supplementation on lymphocyte DNA damage in ICR mice treated with DEN was evaluated using the Comet assay. Mice were divided into 3 groups: control, DEN, and DEN + magnetized water group. Fifteen mice were maintained in each group for the entire experimental period of 6, 12 and 18 weeks. Five mice in each group were sacrificed at 6, 12, and 18th weeks, followed by the Comet assay using the blood obtained from heart puncture of the mice. The level of lymphocyte DNA damage reflected by tail moment and other DNA damage indices of tail DNA (%) or tail length of the magnetized water group were significantly decreased after the 6th, 12th and 18th weeks of supplementation compared with the positive control, the DEN group. The relative DNA damage of the magnetized water groups compared to the DEN control group after 6th, 12th, and 18th weeks of supplementation were 42.2%, 40.8%, and 32.9% for DNA in tail, 31.2%, 32.6%, and 21.3% for tail length, and 33.8%, 33.8%, and 24.6% for tail moment, respectively. This is the first report demonstrating that magnetized water may be involved in the lowering effect of the DNA damage in DEN-treated ICR mice. This result suggests that the magnetized water might have minimized the DNA damage by improving the antioxidant status of the mice. However, further studies are needed to characterize the condition of the magnetization and examine the long-term effect of the water product.