Normal Rat Liver Research Articles

Effect of hyperbaric oxygen therapy on weight loss and hyperlipidemia in rats

ObjectiveTo explore whether hyperbaric oxygen therapy(HBO) can promote weight loss and recovery of hyperlipidemia in rats, and to explore the possible mechanism. Methods180 SD rats were divided into 6 groups with 30 rats in each group. The first 3 groups were intraperitoneally injected with fat emulsion to make hyperlipidemia models, and the last three groups were injected with normal saline. The first three groups received 3 h/d, 6 h/d, 0 h/d HBO therapy respectively, and the last three groups received the same treatment. Body weight, blood lipid and transaminase were measured in all SD rats, and pathological sections of heart, liver and kidney were observed. ResultsHyperlipidemia group treated with 3 h/d hyperbaric oxygen has the effect of reducing hyperlipidemia compared with other groups and has the effect of heart and kidney protection. Although 6 h/d HBO therapy has a more noticeable effect on lowering hyperlipidemia, it has more apparent liver damage effects. The normal group treated with HBO for 3 h/d or 6 h/d both have the effect of weight loss, and the impact of liver injury is not apparent. However, the 6 h/d HBO therapy group had a more prominent weight loss effect. ConclusionHBO therapy can promote weight loss and reduce hyperlipidemia. Our experiments have shown that 6 h/d and 3 h/d HBO therapy reduces blood lipids in hyperlipidemia SD rats. However, the former has noticeable liver damage effects on SD rats, and the latter is adequate for protecting the liver in normal or hyperlipidemia SD rats. At the same time, it has been proved that HBO therapy has cardio and kidney protection in hyperlipidemia SD rats.

ABHRAK BHASMA AND SIO2 INFLUENCED MOBILITY OF LIPIDS IN LIVER AND KIDNEY OF CCL4 INDUCED ACUTELY INTOXICATED ALBINO RAT

In our earlier studies on CCl4 induced acute toxicity model (CCl4 3ml/kg body wt). Abhrak bhasma protects fatty degeneration of liver and associated nephrotoxicity in male albino rats. It had shown to function through production and management of free radicals (Teli and Kanase, 2020a, b). To study further the paths of Abhrak Bhasma mediated protection of acute hepatotoxicity and associated nephrotoxicity, liver, kidney and serum lipid contents were studied in present work. It shows no lipid accumulation in liver or kidney of normal rats by Abhrak Bhasma (10, 20, 30 and 40mg doses). But a same dose of silica in pure form (SiO2) is hepato and nephrotoxic in high doses in normal male albino rat. In acutely intoxicated rat also all the doses of Abhrak Bhasma influenced lipid contents of liver, kidney and serum show the protection of liver from fatty degeneration and also associated nephrotoxicity. Doses 30 and 40mg normalized the contents from liver, kidney and serum. The results are discussed to reveals the probable mode of action of Abhrak Bhasma.

Effect of PINK1 and Parkin gene silencing on sodium arsenite-induced mitophagy in normal rat liver cells (BRL-3A).

NaAsO2-induced liver damage leads to autophagy, which plays an important role in cell quality control. Mitophagy plays an important role in hepatocyte damage, and PINK1 and Parkin constitute an important pathway in mitophagy. PINK1 selectively degrades abnormal mitochondria, and Parkin can recognize damaged mitochondria. However, the mechanism underlying the involvement of PINK1/Parkin in NaAsO2-induced mitophagy is unclear. Transfection plasmids containing dsRNA were used to interfere with the expression of Parkin in the following groups: the empty plasmid group was established by add the empty plasmid only, the PINK1-knockdown (KD) group was established by adding 5μg of PINK1 dsRNA and then by adding 10mM NaAsO2, and the Parkin-KD group was established by adding 5μg of Parkin dsRNA and then by adding 10mM NaAsO2. The expression of PINK1 and Parkin in autophagy was detected by western blotting and immunofluorescence staining. The ultrastructures of autophagosomes and mitochondria were observed by transmission electron microscopy. The successful KD of PINK1 and Parkin aggravated the NaAsO2-induced damage to mitophagy. The degeneration of mitochondrial vacuoles and the appearance of autophagosomes were detected in the NaAsO2, NaAsO2+PINK1-KD and NaAsO2+Parkin-KD groups. NaAsO2 can induce mitophagy in rat hepatocytes, and the silencing of PINK1 and Parkin can aggravate mitochondrial damage during this process. This study explored the mechanism of NaAsO2-induced mitophagy in BRL-3A cells after PINK1 and Parkin gene silencing.

Palm leaf catechins improved insulin-related pathways in diabetic rats

Catechin-rich oil-palm leaf extract (OPLE) (Elaeis guineensis) was previously demonstrated to possess benefits for diabetes and cardio metabolic health (vasodilation, antioxidant, cardiovascular, anti-hypertensive, anti-inflammatory, hepatoprotective and nephroprotective properties) in animal models. For insights into OPLE anti-diabetic mode-of-action and possible toxicity, the effects of dietary OPLE on insulin-signaling pathways mRNA expressions in the liver, kidney, pancreas, and spleen of normal and diabetic rats were examined. Type-2-Diabetes Mellitus (T2DM) were induced by chronic high-fat diet and streptozotocin (35 mg/kg) intraperitoneal injection. The OPLE (100 mg/kg body weight) were fed daily to normal and T2DM-induced rats. The OPLE suppressed hyperglycaemia and excessive weight gain in the T2DM rats, and appeared harmless to normal rats. The OPLE supplementation significantly (p<0.05) modulated the mRNA expressions of phosphatidylinositol-3 kinase (PIK3R1); insulin signaling receptor (INSR); insulin-receptor substrates 1 and 2; and ectonucleotide pyrophosphatase-1 (ENPP1) especially in the livers of normal rats and the spleen of diabetic rats. Results suggested the OPLE probably help prevent diabetes in healthy mammals and ameliorate the immune functions of diabetic mammals. The OPLE improved the antioxidant defence responses, insulin-pathways mRNA expressions in the normal and diabetic rats; suppressed hyperglycaemia and excessive weight gain in T2DM rodents without observable liver or kidney toxicity at the dose used.

Metabolic adaptation to the chronic loss of Ca2+ signaling induced by KO of IP3 receptors or the mitochondrial Ca2+ uniporter

Calcium signaling is essential for regulating many biological processes. Endoplasmic reticulum inositol trisphosphate receptors (IP3Rs) and the mitochondrial Ca2+ uniporter (MCU) are key proteins that regulate intracellular Ca2+ concentration. Mitochondrial Ca2+ accumulation activates Ca2+-sensitive dehydrogenases of the tricarboxylic acid (TCA) cycle that maintain the biosynthetic and bioenergetic needs of both normal and cancer cells. However, the interplay between calcium signaling and metabolism is not well understood. In this study, we used human cancer cell lines (HEK293 and HeLa) with stable KOs of all three IP3R isoforms (triple KO [TKO]) or MCU to examine metabolic and bioenergetic responses to the chronic loss of cytosolic and/or mitochondrial Ca2+ signaling. Our results show that TKO cells (exhibiting total loss of Ca2+ signaling) are viable, displaying a lower proliferation and oxygen consumption rate, with no significant changes in ATP levels, even when made to rely solely on the TCA cycle for energy production. MCU KO cells also maintained normal ATP levels but showed increased proliferation, oxygen consumption, and metabolism of both glucose and glutamine. However, MCU KO cells were unable to maintain ATP levels and died when relying solely on the TCA cycle for energy. We conclude that constitutive Ca2+ signaling is dispensable for the bioenergetic needs of both IP3R TKO and MCU KO human cancer cells, likely because of adequate basal glycolytic and TCA cycle flux. However, in MCU KO cells, the higher energy expenditure associated with increased proliferation and oxygen consumption makes these cells more prone to bioenergetic failure under conditions of metabolic stress.

Concentrations and pharmacokinetic parameters of MRI and SPECT hepatobiliary agents in rat liver compartments

BackgroundIn hepatobiliary imaging, systems detect the total amount of agents originating from extracellular space, bile canaliculi, and hepatocytes. They add in situ concentration of each compartment corrected by its respective volume ratio to provide liver concentrations. In vivo contribution of each compartment to liver concentration is inaccessible. Our aim was to quantify the compartmental distribution of two hepatobiliary agents in an ex vivo model and determine how their liver extraction ratios and cholestasis (livers lacking canalicular transporters) might modify it.MethodsWe perfused labelled gadobenate dimeglumine (Bopta, 200 μM, 7% liver extraction ratio) and mebrofenin (Meb, 64 μM, 94% liver extraction ratio) in normal (n = 18) and cholestatic (n = 6) rat livers. We quantified liver concentrations with a gamma counter placed over livers. Concentrations in hepatocytes and bile canaliculi were calculated. Mann-Whitney and Kruskal-Wallis tests were used.ResultsHepatocyte concentrations were 2,043 ± 333 μM (Meb) versus 360 ± 69 μM (Bopta, p < 0.001). Meb extracellular concentrations did not contribute to liver concentrations (1.3 ± 0.3%). The contribution of Bopta extracellular concentration was 12.4 ± 1.9% (p < 0.001 versus Meb). Contribution of canaliculi was similar for both agents (16%). Cholestatic livers had no Bopta in canaliculi but their hepatocyte concentrations increased in comparison to normal livers.ConclusionHepatocyte concentrations are correlated to liver extraction ratios of hepatobiliary agents. When Bopta is not present in canaliculi of cholestatic livers, hepatocyte concentrations increase in comparison to normal livers. This new understanding extends the interpretation of clinical liver images.

In vivo hepatoprotective effect of Morinda elliptica stem extract against liver fibrosis induced by thioacetamide.

Morinda elliptica L. (Rubiaceae) is a phytomedicinal herb, used to treat gastrointestinal complications in Peninsular Malaysia. The study evaluates the in vivo hepatoprotective activity of ethanolic extract of M. elliptica stem in thioacetamide (TAA) induced liver fibrosis in male Sprague Drawly rats. Thirty adult rats were divided into five groups of six rats each. Rats of the normal control group received intraperitoneal injections (i. p.) of vehicle 10% Tween-20, 5ml/kg, and hepatotoxic group 200 mg/kg TAA three times per week respectively. Three supplementary groups were treated with TAA plus daily oral silymarin (50 mg/kg) or M. elliptica (250 or 500 mg/kg). After 8 weeks of treatment, all rats were sacrificed. Liver fibrosis was assessed by gross macroscopic and microscopic tissue analysis, histopathological, and biochemical analysis. The livers of the TAA treated group showed uniform coarse granules, hepatocytic necrosis with lymphocytes infiltration. Contrary, the livers of M. elliptica treated groups (250 and 500 mg/kg) were much smoother and the cell damage was much lesser. The livers of M. elliptica treated groups rats showed elevated activity of SOD and CAT with a significant decrease in MDA level at p < .0001. The level of liver damage parameters, that is, ALP, ALT, and AST, bilirubin, total protein, and albumin were restored to the normal comparable to silymarin. M. elliptica stem extract significantly promoted normal rat liver architecture with significant perfections in biochemical parameters. The molecular contents of M. elliptica with hepatoprotective influence could be discovered, is the future prospective of this study.

Saikosaponin D Inhibits the Proliferation and Promotes the Apoptosis of Rat Hepatic Stellate Cells by Inducing Autophagosome Formation.

Objective This study aimed to investigate the effects of saikosaponin D (SSd) on the proliferation and apoptosis of the HSC-T6 hepatic stellate cell line and determine the key pathway that mediates SSd's function. Methods Cell viability was detected using the CCK-8 kit. The EdU kit and flow cytometry were used to examine cell proliferation. The Annexin V-FITC/PI double staining kit and flow cytometry were used to examine cell apoptosis. Western blot analysis was performed to analyze the expression levels of LC3, Ki67, cleaved caspase 3, Bax, and Bcl2. Autophagosome formation was detected by LC3-GFP adenovirus transfection. Results SSd inhibits the proliferation and promotes the apoptosis of acetaldehyde-activated HSC-T6 cells. SSd treatment increased the expression of cleaved caspase 3 and Bax but reduced that of Ki67 and Bcl2. The same concentration of SSd barely influenced the growth of normal rat liver BRL-3A cells. SSd upregulated LC3-II expression and induced autophagosome formation. Autophagy agonist rapamycin had the same effect as SSd and autophagy inhibitor 3-methyladenine could neutralize the effect of SSd in acetaldehyde-activated HSC-T6 cells. Conclusions SSd could inhibit the proliferation and promote the apoptosis of HSC-T6 cells by inducing autophagosome formation.

Oxidative Effects of Electromagnetic Radiation on Liver, Lung, Kidney and Heart Tissues of Diabetic and Normal Rats

Oxidative Effects of Electromagnetic Radiation on Liver, Lung, Kidney and Heart Tissues of Diabetic and Normal Rats

Modulation of vascular contraction via soluble guanylate cyclase signaling in a novel ex vivo method using rat precision-cut liver slices.

Fibrotic processes in the liver of non‐alcoholic steatohepatitis (NASH) patients cause microcirculatory dysfunction in the organ which increases blood vessel resistance and causes portal hypertension. Assessing blood vessel function in the liver is challenging, necessitating the development of novel methods in normal and fibrotic tissue that allow for drug screening and translation toward pre‐clinical settings. Cultures of precision cut liver slices (PCLS) from normal and fibrotic rat livers were used for blood vessel function analysis. Live recording of vessel diameter was used to assess the response to endothelin‐1, serotonin and soluble guanylate cyclase (sGC) activation. A cascade of contraction and relaxation events in response to serotonin, endothelin‐1, Ketanserin and sGC activity could be established using vessel diameter analysis of rat PCLS. Both the sGC activator BI 703704 and the sGC stimulator Riociguat prevented serotonin‐induced contraction in PCLS from naive rats. By contrast, PCLS cultures from the rat CCl4 NASH model were only responsive to the sGC activator, thus establishing that the sGC enzyme is rendered non‐responsive to nitric oxide under oxidative stress found in fibrotic livers. The role of the sGC pathway for vessel relaxation of fibrotic liver tissue was identified in our model. The obtained data shows that the inhibitory capacities on vessel contraction of sGC compounds can be translated to published preclinical data. Altogether, this novel ex vivo PCLS method allows for the differentiation of drug candidates and the translation of therapeutic approaches towards the clinical use.

Metabolic study of ginsenoside Rg3 and glimepiride in type 2 diabetic rats by liquid chromatography coupled with quadrupole‐Orbitrap mass spectrometry

RationaleGinsenoside Rg3 and glimepiride have been applied to treat type 2 diabetes (T2DM) because of their good hypoglycemic effects. In this study, the effects of ginsenoside Rg3 acting synergistically with glimepiride were investigated in liver microsomes from rats with type 2 diabetes.MethodsAn in vitro incubation system with normal rat liver microsomes (RLM) and type 2 diabetic rat liver microsomes (TRLM) was developed. The system also included two experimental groups consisting of RLM and TRLM pretreated with ginsenoside Rg3 and glimepiride (named the RLMR and TRLMR groups, respectively). The metabolism in the different groups was analyzed by ultra‐performance liquid chromatography coupled with quadrupole‐orbitrap mass spectrometry (UPLC/Q‐Orbitrap MS).ResultsThe results showed that the concentration of glimepiride increased in RLM and TRLM after treatment with ginsenoside Rg3. Five metabolites (M1–M5) of glimepiride were found, and they were named 3N‐hydroxyglimepiride, hydroxyglimepiride, 1,2‐epoxy ether‐3‐hydroxyglimepiride, 1N‐hydroxyglimepiride and 1N,2C,S,O,O‐epoxy ether‐3‐hydroxyglimepiride. The metabolite of ginsenoside Rg3 was ginsenoside Rh2.ConclusionsAn in vitro incubation system with RLM and TRLM was developed. The system revealed pathways that produce glimepiride metabolites. Ginsenoside Rg3 may inhibit the activity of cytochrome P450 enzymes in vitro. The present study showed that ginsenoside Rg3 and glimepiride may be combined for the treatment of T2DM.

Modulatory effect of methanol extract of Annona muricata stem bark on mitochondrial membrane permeability transition pore in normal rat liver and monosodium glutamate-induced uterine hyperplasia

Uterine fibroids are benign tumors that develop in many women of reproductive age. Surgery is the main approach to treatment while other options are also associated with adverse effects. Studies have shown that certain bioactive agents present in medicinal plants elicit their anti-tumor activity by induction of mitochondrial permeability transition (mPT) opening. This research therefore aimed at investigating the effect of methanol extract of Annona muricata (MEAM) on mPT pore opening in normal and monosodium glutamate-induced uterine hyperplasia using female Wistar rats. Mitochondria, isolated from rat liver were exposed to different concentrations (20, 60, 100, 140 and 180μg/mL) of MEAM. The mPT pore opening, cytochrome c release, mitochondrial ATPase (mATPase) activity and the percentage lipid peroxidation were assessed spectrophotometrically. Histological effects of MEAM on the liver, brain and uterus of normal and MSG-treated rats were investigated. The in vitro results showed a significant induction of mPT pore opening by 2.4, 4.2 and 6.4 folds, release of cytochrome c and enhancement of mATPase activity at 100,140 and 180μg/mL, respectively. However, oral administration of MEAM did not induce mPT pore opening, neither any significant release of cytochrome c nor enhancement of mATPase activity at all the dosages used. However, histological assay revealed the presence of MSG-induced cellular damage and uterine hyperplasia which was ameliorated by MEAM co-administration. These findings suggest that MEAM contains phytochemicals that can ameliorate MSG-induced damage and uterine hyperplasia in rats; however, the mechanism might not be via upregulation of mitochondrial-mediated apoptosis.

A novel irinotecan-lipiodol nanoemulsion for intravascular administration: pharmacokinetics and biodistribution in the normal and tumor bearing rat liver

Colorectal cancer is one of the most common cancers in the United States and treatment options are limited for patients who develop liver metastases. Several chemotherapeutic regimens have been used for transvascular liver-directed therapy in the treatment of colorectal liver metastases without clear evidence of superiority of one therapy over another. We describe the development of a novel nanoemulsion through combining irinotecan (IRI), a first line systemic agent used for the treatment of colon cancer, with lipiodol, an oily contrast medium derived from poppy seed oil, and evaluated its pharmacokinetic and biodistribution profile as a function of portal venous chemoembolization (PVCE) versus transarterial chemoembolization (TACE) delivery. The Tessari technique was used to create a stable emulsion (20 mg IRI mixed with 2 mL lipiodol) with resultant particle size ranging from 28.9 nm to 56.4 nm. Pharmacokinetic profile established through venous sampling in Buffalo rats demonstrate that the area under the curve (AUC0−∞) of IRI was significantly less after PVCE with IRI-lipiodol as compared to IRI alone (131 vs. 316 µg*min/mL, p-value = .023), suggesting significantly higher amounts of IRI retention in the liver with the IRI-lipiodol nanoemulsion via first-pass extraction. Subseqent biodistribution studies in tumor-bearing WAG/Rjj rats revealed more IRI present in the tumor following TACE versus PVCE (29.19 ± 12.33 µg/g versus 3.42 ± 1.62; p-value = .0033) or IV (29.19 ± 12.33 µg/g versus 1.05 ± 0.47; p-value = .0035). The IRI-lipiodol nanoemulsion demonstrated an acceptable hepatotoxicity profile in all routes of administration. In conclusion, the IRI-lipiodol nanoemulsion via TACE showed promise and warrants further investigation as an option for the treatment of metastatic colorectal cancer.

Leaf extract of Momordica charantia: Effects on liver enzymes, duodenum antioxidant enzymes, and disaccharidases with liver and duodenum histology

Background: Momordica charantia (MC) plant rich in phytochemicals, promoting sound health conditions, contains nutritional and nutraceutical components. Its popular in traditional and folk’s medicine, extract is used to treat and remedy diseases. Adverse effects produce toxic symptoms in children and laboratory animals. Aims and Objectives: The study investigated activities of liver enzymes, antioxidant enzymes, and disaccharidases in the duodenum and evaluated the histomorphological changes in the duodenum and liver of normal rats orally administered with aqueous leaf extract of MC. Materials and Methods: Fifteen male Wistar rats weight between 150 g and 200 g used for this experiment were divided into three groups equally. Group A received only water and Groups B and C received 100 and 400 mg/kg body weight of extract orally for 21 days. Liver enzymes, duodenum antioxidant enzymes, and disaccharidases were determined using standard methods. Histology of duodenum and liver was studied with hematoxylin and eosin staining. Results: Observation from statistically compared results showed that catalase, superoxide dismutase, and glutathione in the duodenum reduced significantly. Malondialdehyde only increases in Group C but not significant. Maltase, sucrose, and lactase in the duodenum reduced significantly. Total protein, albumin, alanine aminotransferase, and aspartate aminotransferase in the liver reduced significantly, alkaline phosphatase showed increase not significant for the two doses (P < 0.05). Histological study from duodenum and liver showed some degenerated villi, loss of epithelial cell, and goblet cells in the duodenum, central vein constriction with hepatocyte hypertrophy, degeneration, and sinusoid dilation. Conclusion: MC has health benefits with ability to neutralized free radicals, preserve functions of liver and if care is not taken, it can induce negative histomorphological changes in the duodenum and the liver.

Effect of Uygur medicine maturants on related factors of liver energy metabolism in rats

Objective: To investigate the effects of five Uygur medicine maturants on the related factors of liver energy metabolism in rats. Methods: After 30 days of preparation and intragastric administration, the weight changes of rats were observed. Enzyme-linked immunosorbent assay (ELISA) was used to detect Na[Formula: see text] K[Formula: see text] ATPase, Ca2[Formula: see text] Mg2[Formula: see text] ATPase and succinate dehydrogenase (SDH) activities; hepatic glycogen (HG) and muscle glycogen (MG) content, plasma triiodothyronine (T3), tetraiodothyronine (T4), and thyrotropin (TSH) content were detected. Results: The Kan (blood) quality regulator reduced the weight of rats, the Sapra regulator increased the weight of rats, the Savda maturant group, and the Balham maturant group increased the Na[Formula: see text] K[Formula: see text] ATPase, Ca2[Formula: see text] Mg2[Formula: see text] ATP, SDH activity, HG, MG, T3, T4, and TSH content of rats significantly, while the other groups had no significant effect on the energy metabolism-related factors. Conclusion: Five kinds of Uygur medicine maturants have effects on the related factors of energy metabolism in the liver of normal rats, and their elevation of serum thyroxine level is related, among which the effects of the Savda maturant group and the Balham maturant group are more significant.

Transcription profiling of cadmium-exposed livers reveals alteration of lipid metabolism and predisposition to hepatic steatosis

Cadmium is a ubiquitous environmental toxicant that can cause liver steatosis and nonalcoholic fatty liver disease (NAFLD) on long-term exposure. Sixteen Sprague Dawley rats were randomly divided into two groups, and were administered normal saline and 5mg/(kg·d) cadmium chloride by gavage. In vitro, BRL3A cells, a rat normal liver cell line, were treated with different concentrations of cadmium to verify the sequencing results. The RNA-seq revealed 146 upregulated genes and 127 downregulated genes in the cadmium intervention group. The key genes of lipid metabolism were significantly overexpressed, such as Cyp1a1 and Pla2g2d. The GO enrichment analysis showed that the "sterol biosynthetic process" was the most obvious difference. The KEGG analysis showed that 6 of the top 10 differential pathways were related to lipid metabolism. The expression of the essential genes in BRL3A was consistent with the sequencing results. The protein-protein interaction (PPI) yielded that Cyp1a1 is in the central region of the differentially expressed gene network. In conclusion, the chronic cadmium exposure is still an important environmental health problem with a probable tendency to cause NAFLD. It may possibly act by affecting the lipid metabolism in the liver, especially the synthesis and decomposition of unsaturated fatty acids.

PKM2-Induced the Phosphorylation of Histone H3 Contributes to EGF-Mediated PD-L1 Transcription in HCC.

High expression of programmed death-ligand-1 (PD-L1) in hepatocellular carcinoma (HCC) cells usually inhibits the proliferation and functions of T cells, leading to immune suppression in tumor microenvironment. However, very little has been described regarding the mechanism of PD-L1 overexpression in HCC cells. In the present study, we found epidermal growth factor (EGF) stimulation promoted the expression of PD-L1 mRNA and protein in HCC cells. Inhibition of epidermal growth factor receptor (EGFR) could reverse EGF-induced the expression of PD-L1 mRNA and protein. Subsequently, we also observed that the phosphorylation level of Pyruvate kinase isoform M2 (PKM2) at Ser37 site was also increased in response to EGF stimulation. Expression of a phosphorylation-mimic PKM2 S37D mutant stimulated PD-L1 expression as well as H3-Thr11 phosphorylation in HCC cells, while inhibition of PKM2 significantly blocked EGF-induced PD-L1 expression and H3-Thr11 phosphorylation. Furthermore, mutation of Thr11 of histone H3 into alanine abrogated EGF-induced mRNA and protein expression of PD-L1, Chromatin immunoprecipitation (ChIP) assay also suggested that EGF treatment resulted in enhanced H3-Thr11 phosphorylation at the PD-L1 promoter. In a diethylnitrosamine (DEN)-induced rat model of HCC, we found that the expression of phosphorylated EGFR, PKM2 nuclear expression, H3-Thr11 phosphorylation as well as PD-L1 mRNA and protein was higher in the livers than that in normal rat livers. Taken together, our study suggested that PKM2-dependent histone H3-Thr11 phosphorylation was crucial for EGF-induced PD-L1 expression at transcriptional level in HCC. These findings may provide an alternative target for the treatment of hepatocellular carcinoma.

Induction of mitochondrial-mediated apoptosis by methanol extract of Cajanus cajan

Induction of mitochondrial-mediated apoptosis by methanol extract of Cajanus cajanAlao O Oa,b and Olorunsogo O.Oa aDepartment of Biochemistry, University of Ibadan, Nigeria bDepartment of Biochemistry, Lead City University, Ibadan, Nigeria. *All correspondence: olajualao@yahoo.co.uk +2348034728112Background: Attention has recently focused on mitochondrial-mediated apoptosis as a signaling pathway that is disrupted during carcinogenesis. The chemopreventive effects of dietary phytochemicals can be explored in order to combat conditions emanating from dysregulation of mitochondria-mediated apoptosis. Cajanus cajan is a medicinal plant commonly used in Nigeria for the traditional treatment of many diseases such as jaundice, measles dysentery and tumours. This study aims to investigate the effects of Cajanus cajan on the opening of the mitochondrial membrane permeability transition(MMPT) pore, ATPase activities, liver enzyme activities and immunohistochemical expression of apoptotic biomarkers in rat liver. Methods: Eighteen male Wistar rats (150±2.00 g) were assigned into three groups of six animals each and orally administered corn oil (control), 100 and 200 mg/kg MECC once daily for thirty consecutive days. Mitochondrial membrane permeability transition and ATPase activity were determined spectrophotometrically. Caspase 3, caspase 9 and cytochrome C release were determined by immunohistochemistry.Results: In vitro, at 10 and 30 µg/mL pore opening was not observed , however at 50 and 70 µg/mL MECC induced pore opening by 7 and 13 folds. Similarly, enhancement of ATPase activities was also observed. In the in vivo studies; the MECC induced opening at 200 mg/kg bwt.There was a significant increase in the activities of AST, ALT and ALP and caspase 3, caspase 9 and cytochrome C were activated Conclusion: Methanol extract of Cajanus cajan induced mitochondrial -mediated apoptosis in normal rat liver . Hence Cajanus cajan can be used in the management of conditions associated with dysregulation of apoptosis.Keywords: Mitochondria membrane permeability transition pore : apoptosis and Cajanus cajan

Assessment of Antihyperglycemic Effect and Acute Toxicity of the Aqueous Scorzonera undulata Extract in Rats.

The current study was carried out in order to assess the antihyperglycemic effect of Scorzonera undulata (S. undulata). Backgroud: Scorzonera undulata subsp. deliciosa (DC.) Maire is used in the Moroccan pharmacopeia for managing diabetes. The objective of the study was to evaluate the anithyperglycemic activity in normal and streptozotocin (STZ)-induced diabetic rats as well as the acute toxicity of the aqueous extract of the roots of Scorzonera undulata (S. undulata). This study investigated the effects of the root aqueous extract of Scorzonera undulata (AERSU) at a dose of 20 mg/kg on blood glucose levels in normal and streptozotocin(STZ)-induced diabetic rats and the acute toxicity of AERSU was examined. Histopathological examination, preliminary phytochemical investigation, determination of glycogen content and evaluation of α-amylase were also performed. Both single and repeated oral doses of AERSU (20 mg/kg) produced a significant reduction in blood glucose levels in normal and diabetic rats. Furthermore, repeated oral administration of AERSU during 15 days increased the glycogen content in the liver in both normal and diabetic rats, inhibited α- amylase activity and improved the histological architecture of the liver and pancreas in treated diabetic rats and ameliorated some biochemical parameters such as ALT and AST. In addition, the preliminary phytochemical investigation showed the richness of the roots of S. undulata in certain phytochemicals particularly in polyphenols. AERSU exhibits an evident antihyperglycemic activity. This pharmacological effect may be due to the protection of surviving pancreatic β cells and hepatocytes, effect of α-amylase and the improvement of glycogen storage in the liver. The dose used in this study seems to be free of any toxicity.

MTORC1 and mTORC2 expression in inner retinal neurons and glial cells

The retina is one of the most metabolically active tissues, yet the processes that control retinal metabolism remains poorly understood. The mTOR complex (mTORC) that drives protein and lipid biogenesis and autophagy has been studied extensively in regards to retinal development and responses to optic nerve injury but the processes that regulate homeostasis in the adult retina have not been determined. We previously demonstrated that normal adult retina has high rates of protein synthesis compared to skeletal muscle, associated with high levels of mechanistic target of rapamycin (mTOR), a kinase that forms multi-subunit complexes that sense and integrate diverse environmental cues to control cell and tissue physiology. This study was undertaken to: 1) quantify expression of mTOR complex 1 (mTORC1)- and mTORC2-specific partner proteins in normal adult rat retina, brain and liver; and 2) to localize these components in normal human, rat, and mouse retinas. Immunoblotting and immunoprecipitation studies revealed greater expression of raptor (exclusive to mTORC1) and rictor (exclusive for mTORC2) in normal rat retina relative to liver or brain, as well as the activating mTORC components, pSIN1 and pPRAS40. By contrast, liver exhibits greater amounts of the mTORC inhibitor, DEPTOR. Immunolocalization studies for all three species showed that mTOR, raptor, and rictor, as well as most other known components of mTORC1 and mTORC2, were primarily localized in the inner retina with mTORC1 primarily in retinal ganglion cells (RGCs) and mTORC2 primarily in glial cells. In addition, phosphorylated ribosomal protein S6, a direct target of the mTORC1 substrate ribosomal protein S6 kinase beta-1 (S6K1), was readily detectable in RGCs, indicating active mTORC1 signaling, and was preserved in human donor eyes. Collectively, this study demonstrates that the inner retina expresses high levels of mTORC1 and mTORC2 and possesses active mTORC1 signaling that may provide cell- and tissue-specific regulation of homeostatic activity. These findings help to define the physiology of the inner retina, which is key for understanding the pathophysiology of optic neuropathies, glaucoma and diabetic retinopathy.