In Vivo Histological Study the Potential of Borago officinalis on Spleen and Testis Histo-Architecture on CCL4 Induced Albino Male Mice

History of medicine and plants dates backside to seclude past when herbal treatment was used and be the only answer to all kind of pain and disease. Nowadays, greater prominence is again to use phytotherapy all over the world. Herbal medicine is a traditional or folk medicine that based on the use of plants’ seeds, berries, roots, leaves, barks, flowers and plant extracts for medicinal purposes. This research study focused the line on the potentail of aqueous and methanolic extract of Borago officinalis (Borago; BO) on spleen and testis of albino male mice alone or after interaction between both plant extracts with CCL4 (toxic compound) in comparison to controls group (negative control; without any treatment and positive control; mice treated with CCL4 only). The results indicated the ability of plant extracts to modulate toxic effect resulted from CCL4 treatment.

5-O-Demethylnobiletin Alleviates CCl4-Induced Acute Liver Injury by Equilibrating ROS-Mediated Apoptosis and Autophagy Induction

Polymethoxyflavanoids (PMFs) have exhibited a vast array of therapeutic biological properties. 5-O-Demethylnobiletin (5-DN) is one such PMF having anti-inflammatory activity, yet its role in hepatoprotection has not been studied before. Results from in vitro study revealed that 5-DN did not exert a high level of cytotoxicity on HepG2 cells at 40 μM, and it was able to rescue HepG2 cell death induced by carbon tetrachloride (CCl4). Subsequently, we investigated acute liver injury on BALB/c mice induced by CCl4 through the intraperitoneal injection of 1 mL/kg CCl4 and co-administration of 5-DN at (1 and 2 mg/kg) by oral gavage for 15 days. The results illustrated that treatment with 5-DN attenuated CCl4-induced elevated serum aminotransferase (AST)/alanine aminotransferase (ALT) ratio and significantly ameliorated severe hepatic damage such as inflammation and fibrosis evidenced through lesser aberrations in the liver histology of 5-DN dose groups. Additionally, 5-DN efficiently counteracted and equilibrated the production of ROS accelerated by CCl4 and dramatically downregulated the expression of CYP2E1 vitally involved in converting CCl4 to toxic free radicals and also enhanced the antioxidant enzymes. 5-DN treatment also inhibited cell proliferation and inflammatory pathway abnormally regulated by CCl4 treatment. Furthermore, the apoptotic response induced by CCl4 treatment was remarkably reduced by enhanced Bcl-2 expression and noticeable reduction in Bax, Bid, cleaved caspase 3, caspase 9, and apaf-1 expression. 5-DN treatment also induced the conversion of LC3 and promoted the autophagic flux. Conclusively, 5-DN exhibited hepatoprotective effects in vitro and in vivo and prevented liver fibrosis induced by CCl4.

Potential Hepatoprotective Activity of Super Critical Carbon Dioxide Olive Leaf Extracts against CCl4-Induced Liver Damage

Virgin olive oil has demonstrated its effective activity against oxidative stress. However, data on the bioactive effect of olive leaves or their major constituents on the liver are scarce. The present research work was conducted to evaluate the hepatoprotective effects of supercritical carbon dioxide (SC-CO2) extracts from fresh and dried olive leaves on hepatotoxicity caused by carbon tetrachloride (CCl4) in rat models. For this purpose, healthy albino rats of 180–250 g weight were used. The assessment of biochemical markers was carried out on blood and liver tissue. Then, a histopathological study was carried out on liver tissue. The obtained results showed that fresh and dried olive leaf extracts ameliorate the perturbed biochemical parameters caused by CCl4 treatment. Furthermore, the results registered for the histopathological study are in accordance with the biochemical parameters and the protective capacity of SC-CO2 extracts against DNA damage, indicating that olive leaf extracts helped to improve liver fibrosis caused by CCl4 treatment.

EFEK HEPAPROTEKTIF GULA AREN TERHADAP KARBON TETRAKLORIDA PADA TIKUS

Abstrak Studi ini meneliti efek gula aren, senyawa bioaktif yang diisolasi dari gula aren, terhadap kerusakan hati akibat karbon tetraklorida (CCl4). Tikus diperlakukan secara intraperitoneal dengan 0,5 ml / kg CCl4 dan kelompok hewan yang berbeda menerima 25, 50, 100, dan 200 mg / kg sudar palem. Pada 24 jam setelah perawatan CCl4, kadar aminotransferase serum dan peroksidasi lipid meningkat secara signifikan, sedangkan kadar glutathione hati menurun. Perubahan ini dilemahkan oleh gula aren. Studi histologis menunjukkan bahwa gula aren menghambat peradangan portal, nekrosis sentrizonal, dan hiperplasia sel Kupffer, yang merupakan tiga karakteristik paling umum dari kerusakan hati yang diinduksi CCl4. Tingkat serum dan mRNA ekspresi tumor necrosis factor-α secara nyata meningkat dengan pengobatan CCl4 tetapi ditekan oleh gula aren. Level mRNA dan ekspresi protein diinduksi nitric oxide synthase dan heme oksigenase-1 meningkat secara signifikan pada 24 jam setelah perawatan CCl4. Gula aren melemahkan peningkatan protein dan ekspresi gen diinduksi nitric oxide synthase tetapi menambah peningkatan heme oxygenase-1. Temuan ini menunjukkan bahwa gula aren melindungi hepatosit dari kerusakan oksidatif yang disebabkan oleh CCl4, dan perlindungan ini kemungkinan disebabkan oleh induksi ekspresi HO-1 dan penghambatan mediator proinflamasi. Abstract This study examined the effects of palm sugar, a bioactive compounds isolated from palm sugar, on carbon tetrachloride (CCl4)-induced liver injury. Mice were treated intraperitoneally with 0.5 ml/kg CCl4 and different groups of animals received 25, 50, 100, and 200 mg/kg palm sudar. At 24 h after the CCl4 treatment, the level of serum aminotransferases and lipid peroxidation was significantly elevated, whereas the hepatic glutathione content was decreased. These changes were attenuated by palm sugar. The histological studies showed that palm sugar inhibited the portal inflammation, centrizonal necrosis, and Kupffer cell hyperplasia, which are the three most common characteristics of CCl4-induced liver damage. The serum level and mRNA expression of tumor necrosis factor-α were markedly increased by the CCl4 treatment but suppressed by palm sugar. The mRNA and protein expression levels of inducible nitric oxide synthase and heme oxygenase-1 increased significantly at 24 h after the CCl4 treatment. Palm sugar attenuated the increase in the protein and gene expression of inducible nitric oxide synthase but augmented the increase in those of heme oxygenase-1. These findings suggest that palm sugar protects hepatocytes from the oxidative damage caused by CCl4, and this protection is likely due to the induction of HO-1 expression and the inhibition of the proinflammatory mediators.

4-methylumbelliferone Prevents Liver Fibrosis by Affecting Hyaluronan Deposition, FSTL1 Expression and Cell Localization

4-methylumbelliferone (4MU) is an inhibitor of hyaluronan deposition and an active substance of hymecromone, a choleretic and antispasmodic drug. 4MU reported to be anti-fibrotic in mouse models; however, precise mechanism of action still requires further investigation. Here we describe the cellular and molecular mechanisms of 4MU action on CCl4-induced liver fibrosis in mice using NGS transcriptome, Q-PCR and immunohistochemical analysis. Collagen and hyaluronan deposition were prevented by 4MU. The CCl4 stimulated expression of Col1a and αSMA were reduced, while the expression of the ECM catabolic gene Hyal1 was increased in the presence of 4MU. Bioinformatic analysis identified an activation of TGF-beta and Wnt/beta-catenin signaling pathways, and inhibition of the genes associated with lipid metabolism by CCL4 treatment, while 4MU restored key markers of these pathways to the control level. Immunohistochemical analysis reveals the suppression of hepatic stellate cells (HSCs) transdifferentiation to myofibroblasts by 4MU treatment. The drug affected the localization of HSCs and macrophages in the sites of fibrogenesis. CCl4 treatment induced the expression of FSTL1, which was downregulated by 4MU. Our results support the hypothesis that 4MU alleviates CCl4-induced liver fibrosis by reducing hyaluronan deposition and downregulating FSTL1 expression, accompanied by the suppression of HSC trans-differentiation and altered macrophage localization.

PKCδ Mediates NF-κB Inflammatory Response and Downregulates SIRT1 Expression in Liver Fibrosis

The precise mechanism of hepatic cirrhosis remains largely unclear. In particular, a potential regulatory mechanism by which protein kinase C-delta (PKCδ ) affects profibrogenic gene expression involved in hepatic cirrhosis has never been explored. In the present study, we investigated whether PKCδ activation is involved in liver inflammatory fibrosis in both lipopolysaccharide (LPS)-treated RAW 264.7 and CCl4-treated mice. PKCδ was strongly activated by LPS or CCl4 treatment and consequently stimulated nuclear factor (NF)-κB inflammatory response. Interestingly, the activation of PKCδ negatively regulated sirtuin-1 (SIRT1) expression, whereas PKCδ suppression by PKCδ peptide inhibitor V1-1 or siRNA dramatically increased SIRT1 expression. Furthermore, we showed that the negative regulation of PKCδ leads to a decrease in SIRT1 expression. To our knowledge, these results are the first demonstration of the involvement of PKCδ in modulating NF-κB through SIRT1 signaling in fibrosis in mice, suggesting a novel role of PKCδ in inflammatory fibrosis. The level of NF-κB p65 in the nucleus was also negatively regulated by SIRT1 activity. We showed that the inhibition of PKCδ promoted SIRT1 expression and decreased p65 levels in the nucleus through deacetylation. Moreover, the inactivation of PKCδ with V1-1 dramatically suppressed the inflammatory fibrosis, indicating that PKCδ represents a promising target for treating fibrotic diseases like hepatic cirrhosis.

Biological and Phytochemical Screening of Tephrosia purpurea extract on Chemically Induced Hepatocellular Carcinoma with Reference to Biochemical Parameters

Tephrosia purpurea possesses hepatoprotective activity as evidenced by the significant and dose dependent restoring the activities of entire liver cancer marker enzymes, diminution in tumor incidence, decrease in lipid peroxidation (LPO) and increase in the level of antioxidant enzymes (GSH, CAT, SOD, GPx and GST) through scavenging of free radicals, or by enhancing the activity of antioxidant, which then detoxify free radicals. These factors protect cells from ROS damage in NDEA and CCl4-induced hepatocarcinogenesis. Histopathological observations of liver tissues too correlated with the biochemical observations. Thus, present investigation suggested that the Tephrosia purpurea would exert a chemoprotective effect by reversing the oxidant-antioxidant imbalance during hepatocarcinogenesis induced by NDEA and CCl4. Besides Tephrosia purpurea is very much effective in preventing NDEA-induced multistage hepatocarcinogenesis possibly through antioxidant and antigenotoxic nature, which was confirmed by various liver injury and biochemical tumour markers enzymes. The hepatoprotective activity of aTephrosia purpurea of 50 % ethanolic extract was studied using rats. The animals received a single intraperitoneal injection of N-nitrosodiethylamine 200mg/kg body wt followed by subcutaneous injection of CCl4 in a dose of 3 ml/kg body wt.Tephrosia purpureaextract dose dependently and significantly the increase in serum hepatic enzyme levels after NDEAand CCl4 treatment compared to the toxin control group. The results of this study confirmed the antioxidant and hepatoprotective activity of the Tephrosia purpurea extract against carbon tetrachlorideand N-nitrosodiethylamine induced hepatotoxicity in rats.

Hepatoprotective Effects of Camel Milk and Urine on Carbon Tetrachloride (CCL4) induced Liver Damage

Carbon tetrachloride (CCL4) is a highly toxic chemical agent and is the most commonly used drug to experimentally induce liver damage. Aim of the study the aim is to investigate the possible protective role of both camel milk and urine on CCL4 induced liver damage. Method 20 rates were used in the study and they were classified into 4 different groups. Group I control, Group II CCL4 group, Group III (milk+CCL4), Group IV (urine+ CCL4). Biochemical parameters in addition to liver enzymes were estimated to evaluated the liver damage. Results the CCl4 treatment markedly affected the liver-specific enzyme activities. A significant (P < 0.05) increase in serum AST (135± 13.6 IU/L), ALT (157± 24.3 IU/L), and ALP (1209 ± 2.59 IU/L) activities was observed in the CCl4-treated rats compared with those of the control rats (104 ± 6.9 IU/L, 101±14.6 IU/L, 146.8±11.2 IU/L) respectively. Concluded from this study that Camel urine has protective effect against CCL4 induced liver damage more than Camel milk.

Antifibrotic Effect of Smad Decoy Oligodeoxynucleotide in a CCl4-Induced Hepatic Fibrosis Animal Model

Hepatic fibrosis is the wound-healing process of chronic hepatic disease that leads to the end-stage of hepatocellular carcinoma and demolition of hepatic structures. Epithelial–mesenchymal transition (EMT) has been identified to phenotypic conversion of the epithelium to mesenchymal phenotype that occurred during fibrosis. Smad decoy oligodeoxynucleotide (ODN) is a synthetic DNA fragment containing a complementary sequence of Smad transcription factor. Thus, this study evaluated the antifibrotic effects of Smad decoy ODN on carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice. As shown in histological results, CCl4 treatment triggered hepatic fibrosis and increased Smad expression. On the contrary, Smad decoy ODN administration suppressed fibrogenesis and EMT process. The expression of Smad signaling and EMT-associated protein was markedly decreased in Smad decoy ODN-treated mice compared with CCl4-injured mice. In conclusion, these data indicate the practicability of Smad decoy ODN administration for preventing hepatic fibrosis and EMT processes.

Anti-fibrotic effect of Smad Decoy Oligodeoxynucleotide in CCl4-Induced Hepatic Fibrosis Animal Model

Hepatic fibrosis is the wound-healing process of chronic hepatic disease that leads to end-stage of hepatocellular carcinoma and demolition of hepatic structures. EMT has been identified to phenotypic conversion of the epithelium to mesenchymal phenotype that occurred during fibrosis. Smad decoy oligodeoxynucleotide (ODN) is a synthetic DNA fragment containing complementary sequence of Smad transcription factor. Thus, this study evaluated the anti-fibrotic effects of Smad decoy ODN on carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice. As shown in histological results, CCl4 treatment triggered hepatic fibrosis and increased Smad expression. On the contrary, Smad decoy ODN administration suppressed fibrogenesis and EMT process. The expression of Smad signaling and EMT-associated protein was markedly decreased in Smad decoy ODN treatment mice compared with CCl4-injuried mice. In conclusion, these data indicate the practicability of Smad decoy ODN administration for preventing hepatic fibrosis and EMT processes.