Phenylhydrazine-induced Anemia Research Articles

Haematological effect of a bi-herbal composition on phenyl hydrazine-induced anaemia in Albino Wistar rats

Picralima nitida and Cymbopogon citratus are plants known for their anti-inflammatory, antioxidant, and haematopoietic properties. This study aimed to determine the haematological effect of a dual herb composition of these plants on phenylhydrazine-induced anaemia in albino Wistar rats. Sixty adult male rats were divided into six groups: control, phenylhydrazine group, ferrous sulphate group, and three groups receiving phenylhydrazine plus 100 mg/kg, 200 mg/kg, and 400 mg/kg of the extract. Haematological parameters were measured using an autoanalyzer and analyzed with SPSS software V22. The red blood cell count was significantly higher in the control group compared to other groups (p<0.05). Haemoglobin was significantly higher in the control, ferrous sulphate, 100 mg/kg, and 400 mg/kg groups compared to the phenylhydrazine and 200 mg/kg groups (p<0.05). Mean cell volume was significantly lower in the control group compared to all other groups (p<0.05). Mean cell haemoglobin was significantly lower in the control group compared to other groups while MCHC was higher in the control group compared to phenylhydrazine, ferrous sulphate, 100mg/kg and 200mg/kg groups (p<0.05). RDWSD was significantly lower in the control group compared to all other groups (p<0.05). Total white blood cell count was significantly lower in the control and phenylhydrazine groups compared to others (p<0.05). Lymphocyte was significantly higher in the ferrous sulphate group compared to others, while monocyte count was significantly lower in the control, phenylhydrazine, and ferrous sulphate groups (p<0.05). Neutrophil was significantly higher in the control and phenylhydrazine groups (p<0.05). Platelet was significantly lower in the control group compared to ferrous sulphate, 200 mg/kg, and 400 mg/kg groups (p<0.05). Mean platelet volume was significantly lower in the control, phenylhydrazine, ferrous sulphate, and 100 mg/kg groups compared to the 400 mg/kg group (p<0.05). The study concludes that the biherbal formulation of the plants caused significant alterations in haematological parameters.

Ameliorative Effect of Pomegranate Molasses on Phenylhydrazine-Induced Anemia in Rats

Ameliorative Effect of Pomegranate Molasses on Phenylhydrazine-Induced Anemia in Rats

Whole Blood Aggregometry in Mice.

Aggregometry plays a crucial role in both clinical diagnostics and research within hematology, serving as a fundamental tool for understanding platelet function and its implications in physiological and pathological processes. In research, aggregometry provides insights into platelet aggregation dynamics and aids in understanding the underlying mechanisms of hemostasis, thrombosis, and related disorders. Light transmission aggregometry (LTA) and lumi-aggregometry, as well as whole blood aggregometry, are commonly employed methods. While LTA and lumi-aggregometry allow for specific platelet function assessment under controlled conditions, whole blood aggregometry provides a more physiologically relevant approach by evaluating platelet aggregation within the context of whole blood. Although both methodologies offer unique advantages, whole blood aggregometry allows for preservation of the native cellular environment, simplicity, and potential for better clinical correlation. In a clinical setting, with human blood samples, protocols are established for both LTA and whole blood aggregometry as they are frequently used diagnostic tools. A protocol for LTA and lumi-aggregometry in murine models has been described; however, to date, there is no standardized protocol for whole blood aggregometry in murine models accessible to hematology researchers. This article aims to outline a simple, basic protocol for murine whole blood aggregometry, offering an alternative method to the commonly used LTA aggregometry in research settings. Standardizing whole blood aggregometry protocols in murine models could enhance experimental reliability and facilitate translational research efforts in hematology. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Whole blood aggregometry in mice Support Protocol: Phenylhydrazine-induced anemia in wild-type mice Basic Protocol 2: Hematocrit percentage in mice.

Unveiling the Antianemic Activity, Physicochemical Aspects, Antioxidant Properties, and Mineral Profile of Petroselinum crispum L.

Anemia is a widespread global health concern necessitating effective, accessible, and natural interventions. The potential of medicinal plants to address anemia has garnered significant interest. Among these plants, parsley (Petroselinum crispum (Petroselinum crispum) L.) stands out as an edible and herbal-based option for combating anemia. Aim of the study: This study investigated the potential of P. crispum (PC-Ext) as an emerging antianemic product, focusing on its physicochemical attributes, antioxidant properties, and mineral profile. Both qualitative and quantitative analyses of the phenolic compounds in P. crispum were conducted by using high-performance liquid chromatography with a diode array detector (HPLC-DAD). Anemia was induced in rats by intravenous injections of phenylhydrazine, administered at a dose of 40 mg/kg for two consecutive days. The antianemic activity of PC-Ext was assessed at a dose of 500 mg/kg twice daily for 5 weeks by estimating blood parameters, such as serum iron and ferritin. Additionally, the osmotic fragility test measured the capacity of red blood cells to withstand osmotic shock of various concentrations of saline. Aqueous extract of P. crispum was rich in phytochemical compounds, including syringic acid, quercetin, catechin, gallic acid, and luteolin. The findings demonstrate the effectiveness of P. crispum in ameliorating phenylhydrazine-induced reductions in red blood cell count (RBCs), hemoglobin (Hb), and hematocrit (HCT) levels. Consequently, PC-Ext exhibits significant activity against phenylhydrazine-induced anemia in rats, as demonstrated by its ability to prevent hemolysis. Iron estimation within PC-Ext further confirms its utility in addressing both iron deficiency and ferritin-deficiency anemia. Therefore, PC exhibits a favorable effect against both types of anemia, iron deficiency, and hemolysis. The results of this study provide robust scientific validation for ethnomedicinal use and the potential utility of P. crispum, positioning it as a promising source for future pharmaceutical development.

Antianemic Activity of Allolobophora caliginosa Extract against Phenylhydrazine-induced Anemia in Rats

Background:: Anemia is a global hematological disorder. Earthworms have antimicrobial, antispasmodic, antihypertensive, antiallergic, anti-inflammatory, and anticoagulant properties. Objective:: Our study aimed to investigate the anti-anemic activity of the earthworm Allolobophora caliginosa extract (AcE) on phenylhydrazine-induced hemolytic anemia in rats. Methods:: Eighteen adult male albino rats were randomly divided into 3 groups (n = 6 per group) as follows: Control, PHZ, and AcE-treated groups. Induction of anemia was done by intraperitoneal administration of phenylhydrazine (40 mg/kg) for 2 consecutive days. Treatment of AcE by intraperitoneal injection (45 mg/kg) for six successive days. Results:: Treatment of anemic rats with AcE extract caused significant increases in red blood cells (RBCs), hemoglobin (HB), hematocrit (HCT), total protein (TP), albumin, glutathione reduced (GSH), catalase (CAT), and glutathione-S-transferase (GST), as compared to the PHZ group. While, mean corpuscular haemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), malondialdehyde (MDA), and nitric oxide (NO) levels significantly decreased after intraperitoneal administration of AcE, when compared to the PHZ group. Liver sections from the AcEtreated group restored the normal micromorphological features of the liver. The expression of caspase- 3 was weak and/or there was no expression in the liver sections from AcE groups. AcE caused a significant reduction in the incidence of DNA damage, which appeared as less DNA in the tails. Conclusion:: The present research sheds light on the antianemic efficacy of AcE in rats. This study reveals that AcE has antianemic activity, which can be linked to its iron and branched-chain amino acid content, as well as antioxidant and anti-apoptotic properties.

Evaluation of hematinic effects of aqueous leaf extract of hibiscus acetosella welw. Ex hiern on phenyl hydrazine-induced anemia in wistar rats

Ethnopharmacological information indicates that the leaves of Hibiscus acetosella Welw. ex. Hiern are used in the treatment of anemia in western and central Uganda. In this study, the aqueous leaf extract of Hibiscus acetosella Welw. ex. Hiern was tested for its haematinic activity in Wistar rats. Phenylhydrazine (PHZ- 4 mg/kg, IP) was used to induce anemia in rats for a period of two weeks. Animals were grouped into six groups (1, 2, 3, 4, 5, and 6); each group had six rats. Group 1 (negative control), group 2 (positive control), group 3 were treated with FeSO4, and groups 4, 5, and 6 were treated with different doses of the extract (1000 mg/kg, 2000 mg/kg, and 4000 mg/kg, respectively). Screening of phytochemicals was performed, and acute toxicity study was carried out according to the Lorke's experiment 1993. Phytochemical analysis revealed that the aqueous leaf extract of Hibiscus acetosella Welw. ex. Hiern contains tannins, flavonoids, diterpenes, steroids, and saponins. The aqueous leaf extract of Hibiscus acetosella Welw. ex. Hiern exhibits haematinic effects. Keywords: Hematinic effects, Aqueous leaf extract, Hibiscus acetosella Welw. ex Hiern, Phenyl hydrazine induced anemia, and Wistar rats.

Anti-Anemic Activity of Raphanus sativus Leaves on Phenylhydrazine Induced Anemia in Wistar Albino Rats

Anti-Anemic Activity of Raphanus sativus Leaves on Phenylhydrazine Induced Anemia in Wistar Albino Rats

Anti-anemic potential of moringa oleifera flower extract against phenylhydrazineinduced anemia in rats

Anti-anemic potential of moringa oleifera flower extract against phenylhydrazineinduced anemia in rats

Ameliorative Effects of Coconut Water on Hematological and Lipid Profiles of Phenylhydrazine-treated Rats.

Anaemia is a widespread health issue affecting young children and pregnant women, characterized by reduced red blood cells or haemoglobin levels. Coconut water, rich in nutrients such as L-arginine, iron, vitamin C, vitamin B6, folic acid and fatty acids, is believed to aid in blood formation (hematopoesis). The study aimed to examine the impact of coconut water on hematological indices and lipid profiles in rats with phenylhydrazine-induced anemia. 30 rats were divided into 5 groups: a normal control, phenylhydrazine untreated, coconut water (0.5ml/kg), iron, and ferrous treated groups. Hemoglobin, hematocrit, and erythrocyte levels were measured using a Hematology Analyzer. Results showed a significant decrease in LDL and TG levels, and an increase in HDL levels in phenylhydrazine induced anemia compared to the control group. Coconut water administration at 0.5ml/kg reduced LDL, VLDL and TG levels, and increased HDL levels in rats with induced anemia. The study found that coconut water had a positive effect on hematological indices, as it increased hemoglobin and erythrocyte levels in rats with induced anemia. These findings suggest that coconut water may have potential therapeutic benefits for individuals with anemia, particularly in lowering lipid levels and improving blood formation. However, further research is needed to fully understand the mechanisms underlying these effects and to determine the most effective dosage and duration of treatment. Overall, the study highlights the importance of coconut water as a potentially beneficial alternative treatment for anemia.

M6A Mediated Ribostasis of RNA Stress Granule Assembly Governs Blood Development and Regeneration

Hematopoietic stem cell (HSC) form diverse cell types and requires transcription and translation regulation. Using CD34+ human hematopoietic stem and progenitor cells (hCD34+) combined with N6-methyladenosine (m6A) sequencing, we identified a novel regulatory mechanism via stress granule regulation during erythropoiesis. Our single-nucleotide resolution m6A analysis of the whole transcriptome identified ~4800 transcripts to be m6A modified and showed demethylation (>2-14 fold) during erythropoiesis. Genome-wide CHIP-seq analysis of hCD34+ cell erythropoiesis with anti-Gata1 showed an enhancer enrichment at the ALKBH5 RNA demethylase gene. Using ALKBH5 CRISPR Cas9 KO strategy combined with hCD34+ cell in-vitro differentiation protocol we observed a block in hCD34+ cell erythropoiesis thus validating Alkbh5 role in erythropoiesis. hCD34+ ALKBH5 KO cells showed a failure of RNA m6A demethylation and discovered stress granule (SGs) transcripts m6A levels to be upregulated (>2-4 fold, p=0.01). Stress granules are membrane-less organelles composed of various RNA binding proteins. SGs form a hub in the cytoplasm that governs RNA metabolism and global proteome by translational regulation. Our analysis showed ATXN2, G3BP1/2, Tia-1,and PABPC1 SGs transcripts are m6a modified at 3'UTR. K562 cell ALKBH5 KO erythropoiesis model led to massive m6A upregulation on ATXN2 3'UTR (>14 fold, p=0.001) and other SGs transcripts. Mass spectrometry analysis of hCD34+ ALKBH5 KO cell proteome showed downregulation of ATXN2(>2 fold, -log P value2), confirming 3' UTR m6A mediated protein synthesis regulation. ALKBH5 KO hCD34+ cells showed aggregation of SGs protein in the cytoplasm due to altered SG protein stoichiometry(>4-10 fold, p=0.001). Zebrafish alkbh5 KO showed defective blood development and anemia. STED microscopy of the whole kidney marrow of alkbh5 KO fish showed the presence of G3BP and ATXN2 SGs with m6A labeled RNAs. alkbh5 KO zebrafish blood and kidney marrow analysis also showed increased myeloid and lymphoid lineages indicating early lineage bias. We used the alkbh5 KO zebrafish model and subjected it to phenyl hydrazine-induced anemia to study its role in stress-induced regeneration. Using FACS analysis of whole kidney marrow, we show red cell recovery failure in alkbh5 KO zebrafish models. Temporal analysis of alkbh5 KO marrow cells during PHZ-induced acute anemia and regeneration showed a systemic failure to resolve SGs compared to controls. We confirmed these results in the mouse ALKBH5 KO model establishing the role of ALKBH5 in stress erythropoiesis. Furthermore, hCD34+ ATXN2 KO combined with Methocult-colony formation assay showed defects in erythroid and myeloid lineages. Super-resolution microscopy of hCD34+ ATXN2 KO cells phenocopied the presence of disease-associated ALS (amyotrophic lateral sclerosis) stress granules with TDP-43, G3BP1/2, PABPC1, and Tia-1 enriched with m6A labeled RNAs. Using the K562 ALKBH5 KO erythropoiesis model and ATXN2 lentiviral overexpression, we effectively resolved stress granules and blocks in erythropoiesis. In summary, m6A marks on ATXN2 mRNA regulate SG biology, affecting HSC differentiation and impacting blood development. SG-mediated regulation of HSC fate may highlight new therapeutic strategies for blood disorders.

Anti-anaemic effects of leaf protein concentrate of Eremomastax speciosa (Hochst) Cufod. on some blood parameters of male Sprague-Dawley rats exposed to phenylhydrazine-induced anaemia

This study investigated the effects of leaf protein concentrate (LPC) of Eremomastax speciosa (Hochst.) Cufod on some haematological and serum profile of 45 male Sprague-Dawley rats divided into nine groups of five rats per group. The rats received fresh drinking water daily, with feed at 85% of ad-lib. The standard haematinic used was Ranferon-12®, at the dose of 0.3ml/kg body weight. The resultant groups were; Group1 (Positive control, received commercial feed) Group 2 (phenylhydrazine only or negative control), Group 3 (anaemia +haematinic), 4, 5 and 6 (anaemia +100 mg/kg, 200 mg/kg and 400 mg/kg of LPC respectively), Groups 7, 8, 9 (Untreated (or UT) +100 mg/kg, 200 mg/kg and 400 mg/kg of LPC respectively). The LPC was administered orally to the designated groups, while the controls received distilled water in place of the LPC. Anaemia was induced by intraperitoneal injection of phenylhydrazine at 40 mg/kg for 2 days. Samples for haematological and biochemical analysis were collected on days 9, 21 and 28 of the experiment. Results showed that the groups supplemented with LPC were better in terms of the haematology and serum biochemical values compared to the negative control, and this strongly supported the traditional use of E. speciosa leaf in the treatment of anaemia.

Haematinic and Hepatoprotective Properties of Telfairia occidentalis Fruit Mesocarp on Phenylhydrazine-Induced Anaemia in Experimental Rats.

The level and potential of iron contained in fluted pumpkin (Telfairia occidentalis) has been exploited as a blood tonic; however, the potentials of some other parts of the plant are unknown. The effect of T. occidentalis fruit mesocarp (aqueous extract) on phenylhydrazine (PHZ)-induced anaemia in experimental rats was investigated in a bid to determine its curative properties and potential in reversing haemolytic anaemia and protection of liver health. The LD50 of the fruit extract was determined using Lorke's method for the determination of acute toxicity. The study involved oral administration of varying doses of the extract to different groups of rats which were monitored for 24 hours. The test sample did not show any signs of toxicity at doses of 5000 mg/kg b.wt, which is the highest possible recommended dose for toxicity testing. For the evaluation of the effects of the fruit extract on haematological indices and biochemical enzyme markers in anaemic rats, 30 matured albino Wistar rats were used. The rats were divided into five groups of six rats each. Group 1 consisted of normal rats (control group), Group 2 consisted of anaemic untreated rats, and Group 3 consisted of anaemic rats treated with the standard drug Astymin, while Groups 4 and 5 were made up of anaemic rats given the extract at doses of 600 mg/kg b.wt and 1000 mg/kg b.wt, respectively. The fruit extract failed to show any significant effect in improving the haemoglobin (Hb), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphate (ALP) levels in anaemic rats but rather may have contributed to a reduction in Hb levels and an unhealthy increase in serum enzyme levels. This is indicative of the apparent inability of the aqueous extract of the T. occidentalis fruit mesocarp to reverse PHZ-induced haemolytic anaemia and may suggest a possible detrimental effect of high doses of the extract over a prolonged period.

Anti-Anemic Potential of a Herbal Recipe Against Phenylhydrazine-Induced Anemia in Rats

Anemia is characterized by an abnormal decrease in the level of hemoglobin or the number of red blood cells in the blood. It is a public health problem that mainly affects pregnant women and young children. In the present study, the objective was to evaluated the anti-anemic potential of a recipe composed of three medicinal plants, <i>Xylopia aethiopica </i>(Dunal) A. Rich<i>, Zanthoxylum leprieurii (GUILL) </i>and<i> Harungara madagascariensis (LAM), </i>used to treat sickle cell anemia in rats. Aqueous and hydroethanol extracts of recipe composed were first prepared and quantitative phytochemical tests were carried out. Then, the anti-anemic activity of the recipe extracts was assessed. The anemia was induced in animals by intraperitoneal injection of phenylhydrazine at a repeated dose of 40 mg/kg bw for two days. Treatment was carried out with the aqueous and hydroethanol extracts of the plant recipe at doses of 400 and 800 mg/kg bw. The obtained results indicated that recipe composed contains total phenols (43.31±0.92 mg GAE/g and 33.82±0.6 mg GAE /g of dry weight), total flavonoids (17.07±0.89 mg QE and 19.76±1.15 mg QE/g of dry weight) and total tannin (8.65±0.01 mg QE and 26.5±0.95 mg Cat E/g of dry weight) in decocted and hydroethanol extracts respectively. The anemia was effectively corrected after 21 days of oral administration of the extracts to the rats. The extracts significantly increased hemoglobin (P < 0.05), red blood cell count (P < 0.001), and hematocrit (P < 0.01) as early as the 7^th days of treatment compared with untreated controls. These results show that the plant recipe composed of <i>Xylopia aethiopica, Zanthoxylum leprieurii </i>and<i> Harungara madagascariensis </i>possess antianemic properties.

The hematopoietic potential ofmethanolic and aqueous extracts of Portulaca oleracea in a phenylhydrazine model of anemia.

Portulaca oleracea, commonly known as Purslane, is traditionally used as a sour, diuretic, and cooling herb with hemostatic properties. The present study evaluates the antianemic effect of methanolic and aqueous extracts of P. oleracea in a phenylhydrazine model of anemia. Phenylhydrazine (60 mg/kg/day, i.p., two consecutive days) was used to induce anemia in rats. The aqueous and methanolic extracts of P. oleracea were prepared, and three methods of treatment were defined with two doses (500 and 750 mg/kg, i.p.). The hematological parameters and blood cell morphology, total and direct bilirubin, and morphology, and pathology of bone marrow were evaluated. The results showed that the methanolic extract has better effects than aqueous extract in improving phenylhydrazine-induced anemia. Our results showed that administration of 500 and 750 mg/kg of P. oleracea methanolic extracts for 4 days could protect against the development of anemia caused by phenylhydrazine. In summary, the methanolic extracts of P. oleracea might be effective in phenylhydrazine-induced anemia.

Terminalia catappa attenuates phenylhydrazine-induced anaemia and hepato-renal toxicity in male Wistar rat by boosting blood cells, modulation of lipoproteins and up-regulation of in vivo antioxidant armouries

Aim This study investigated the haematinic, antihyperlipidaemic, hepato-renal protective effects of Terminalia catappa aqueous leaf extract on male Wistar rats exposed to phenylhydrazine toxicity. Methods Animals were exposed to phenylhydrazine (PHZ) 50 mg/kg intraperitoneal for two consecutive days thereafter, treated with T. catappa extract (100 mg/kg and 200 mg/kg) orally for 21 days. After the experimentation, animals were sedated with ketamine (70 mg/kg) and euthanized by cervical dislodgement. Blood and organs were collected for haematology and biochemical studies following standard laboratory methods. Results Our study showed that T. catappa significantly increased erythrocytes, haemoglobin, haematocrit and high density lipoprotein as well as down-regulating leukocytes, thrombocytes, ALP AST, ALT creatinine, urea, total cholesterol as well as low density lipoprotein. The liver, kidney and spleen antioxidant defence were also up-regulated against the adverse effect caused by phenylhydrazine exposure. Conclusion Terminalia catappa attenuated Phenylhydrazine-induced anaemia and hepato-renal toxicity in male Wistar rat by boosting blood cells, modulation of lipoproteins and up-regulation of in vivo antioxidant armouries.

Antioxidant and Anti-Anemic Effects of Ethanol Leaf Extracts of Mucuna poggei and Telfairia occidentalis in Phenyl-Hydrazine-Induced Anemia in Wistar Albino Rats

Abstract Introduction With the use of recent therapeutic data, several methods for finding medications from untapped natural materials continue to advance research and development. The antioxidant and antianemic effects of ethanol leaf extracts of Telfairia occidentalis and Mucuna poggei in phenylhydrazine-induced anemia in Wister albino rats were investigated. Method A total of 54 male albino rats were randomly assigned to nine experimental groups (n = 6). Anemia was induced in groups 2 to 9 with 10 mg/kg body weight of phenylhydrazine. Group 1 (normal control) rats were administered normal saline only. Group 2 (anemic control) rats were untreated. Group 3 (standard controls) rats were treated with a standard multivitamin, groups 4, 5, and 6 rats were treated with varying doses of M. poggei leaf extract respectively, while groups 7, 8, and 9 rats were treated with similar doses of T. occidentalis leaf extract by oral intubation for 21 days. Results Earlier, the gas chromatography-mass spectrometry analysis indicated eighteen (18) and twenty-nine (29) active compounds present in M. poggei and T. occidentalis, respectively. More so, T. occidentalis and M. poggei were studied for their inhibitory potentials against 2,2-diphenyl-1 picrylhydrazyl-free radicals, nitric oxide, phosphomolybdenum, and ferric-reducing antioxidant power. M. poggei and T. occidentalis also showed the following trend of minerals Mg &gt; Ca &gt; Fe &gt; Na &gt; Zn &gt; Mn &gt; K and Mg &gt; Fe &gt; Ca &gt; Na &gt; K &gt; Mn &gt; Zn, respectively. The results also showed that the packed cell volume, red blood cells, and hemoglobin significantly (p &lt; 0.05) decreased in anemic control rats compared with normal controls and were significantly (p &lt; 0.05) increased on treatment with the leaf extracts. Catalase, superoxide dismutase, glutathione peroxidase activities, and glutathione levels significantly (p &lt; 0.05) decreased in anemic control, compared with normal. However, these significantly (p &lt; 0.05) increased on treatment with the leaf extracts in a dose-independent manner. Malondialdehyde levels took the reverse trend. Conclusion This study indicated that leaf extracts of M. poggei and T. occidentalis can be used in the management of anemia and oxidative stress-related complications

Intermittent Caloric Restriction Promotes Erythroid Development and Ameliorates Phenylhydrazine-Induced Anemia in Mice.

BackgroundDietary restriction has a profound effect in altering immune system and promoting metabolic health and aging. However, how dietary restriction impacts erythroid system is largely unknown. We found that a short-term caloric restriction (CR) stimulates expression of KLF1, a master regulator of erythroid development, in the spleen of mouse, and thus explored the potential effect of CR on erythropoiesis.MethodsWe analyzed the effects of intermittent CR and continuous CR for different lengths of time on parameters of peripheral blood and erythroid profiles in the spleen and bone marrow in C57BL/6 mice. We next assessed how different types of CR affect phenylhydrazine-induced anemia in the mice. Colony formation assay was also used to analyze LK + progenitors and BFU-E in the bone marrow.ResultsIntermittent CR for 2 weeks raised the number of reticulocytes in the blood, while continuous CR for 2 weeks elevated red blood cells and hemoglobin level. Intermittent CR for 2 weeks promoted extramedullary hematopoiesis in the spleen, while continuous CR mainly promoted erythropoiesis in the bone marrow. Interestingly, a short-term intermittent CR but not continuous CR was able to ameliorate phenylhydrazine-induced anemia. Intermittent CR reduced early-stage erythroblasts and increased late-stage erythroblasts/mature RBCs in the spleen, indicating an accelerated transition from early-stage to late-stage erythroblasts/mature red blood cells. Furthermore, a short-term intermittent CR elevated LK + progenitors and the committed erythroid progenitor cells BFU-E in the bone marrow.ConclusionOur study demonstrated that a short-term intermittent CR, but not continuous CR, has a significant effect to promote hematopoiesis and such activity can ameliorate phenylhydrazine-induced acute anemia in the mouse.

Phenylhydrazine-induced anaemia reduces subcutaneous white and brown adipose tissues in hypothalamic obese rats.

What is the central question of this study? Can an anaemic state modify adiposity and metabolic parameters in hypothalamic obese rats? What is the main finding and its importance? Hypothalamic obese rats do not display iron deficiency. However, the pharmacological induction of anaemia in hypothalamic obese rats resulted in reduced adiposity, characterized by a decrease in subcutaneous white and brown adipose tissue depots. These findings suggest that iron imbalance in obesity may elevate lipolysis. Iron imbalance is frequent in obesity. Herein, we evaluated the impact of anaemia induced by phenylhydrazine on adiposity and metabolic state of hypothalamic obese rats. Hypothalamic obesity was induced by high doses of monosodium glutamate (MSG; 4g/kg) administered to neonatal male rats (n=20). Controls (CTL; non-obese rats) received equimolar saline (n=20). Rats were weaned at 21days of life. At 70days, half of the rats received three intraperitoneal doses of phenylhydrazine (PHZ; 40mg/kg/dose) or saline solution. Body weight and food intake were followed for 4weeks after PHZ administration. At 92days, rats were killed and blood was collected for microcapillary haematocrit (Hct) analysis and plasma quantification of glucose, triglycerides, total cholesterol and iron levels. The liver, the spleen, and the white (WAT) and brown (BAT) adipose tissues were excised, weighed and used for histology. MSG-treated rats developed obesity, hypertriglyceridaemia and insulin resistance, compared to CTL rats, without changes in iron levels and Hct. PHZ administration reduced plasma iron levels and promoted similar tissue injuries in the spleen and liver from MSG and CTL rats. However, in MSG-treated rats, PHZ decreased fasting glucose levels and Hct, as well as diminishing the subcutaneous WAT and BAT mass. Although MSG-obesity does not affect plasma iron levels and Hct by itself, PHZ-induced anaemia associated with obesity induces a marked drop in subcutaneous WAT and BAT mass, suggesting that iron imbalance may lead to increased lipolytic responses in obese rats, compared to lean rats.

Study of Therapeutic Effects of Olive Leaf Extract on Phenylhydrazine-Induced Anemia Mice Model

IntroductionAnemia is the most common blood disorder which affects billions of people, especially young adults and women. However, currently available treatments of anemia are with limitations and adverse effects. Therefore, natural resources have been receiving considerable attention as complementary or alternative hematinic agents in recent years. In this regard, olive leaf extract (OLE) is rich in bioactive phenolic compounds and has been reported to have anti-inflammatory, antioxidant and neuroprotective effects (Vogel et al., 2015). Our previous study showed treatment with water extract of olive leaf (WOL) for 12 days could induce erythroid differentiation in human hematopoietic stem cells (hHSCs) and showed potential in oxygen and iron homeostasis, haem metabolism, and hemoglobin (Hb) biosynthesis (Kondo et al., 2021). In the present study, we aimed to investigate the therapeutic effects of WOL on the phenylhydrazine (PHZ)-induced mice model of anemia and explore its underlying molecular mechanism.MethodsTotal 63 male ICR mice (four weeks old) were randomly divided into three groups: control, PHZ and WOL groups. Mice in the WOL group were orally administered with 150 mg/kg body weight of WOL (diluted by saline) every day, while mice in the other two groups received saline. After two weeks of WOL pretreatment, PHZ was injected intraperitoneally (60 mg/kg body weight) to PHZ and WOL groups. The mice were dissected on day0 (before PHZ injection), day1, day3, day5, and day7 after PHZ injection. We collected blood for hematologic tests during dissection and liver, kidney, intestine, bone marrow, and spleen samples. We next checked the iron homeostasis-related gene expressions by real-time PCR: hepcidin (Hamp) in liver, ferroportin (Fpn) in spleen and intestine.Results and DiscussionAfter anemia was induced, WOL group showed a significant increase in the number of reticulocytes, the erythroid progenitors, compared to that of in PHZ group on Day 5. Simultaneously, plasma iron level on Day 5 and Hb level on Day7 was significantly decreased in the WOL group compared to the PHZ group. These results suggest that during anemia, the WOL group had rapid erythropoiesis, which in turn caused a rapid consumption of Hb and iron.Additionally, mRNA expression of Hamp in liver was significantly decreased on Day 5, whereas Fpn expressions in spleen and intestine were significantly increased on Day 5 and Day 7 in the WOL group compared to the PHZ group.Hamp regulates plasma iron concentrations as well as systemic iron metabolism by interacting with its receptor Fpn, a transmembrane iron-exporter. In intestine, Fpn controls iron absorption from food intake, while in spleen, Fpn regulates phagocytosis of senescent erythrocytes by the macrophages. Decreased Hamp expression and increased Fpn expression by WOL treatment suggest increased levels of absorbed and recycled iron to meet the demand for erythropoiesis. Altogether, our findings indicate that WOL could regulate iron homeostasis in the early stage of anemia to promote the differentiation of erythroid progenitors.ConclusionOur study suggests that WOL treatment downregulates Hamp expression in liver and increased Fpn expressions in the intestine and spleen, leading to increased absorbed iron in the intestine and recycled iron in the spleen (Figure 1). Therefore, WOL may have therapeutic potential in anemia through regulating iron homeostasis and promoting erythroid differentiation. However, further studies are required to confirm these findings and to elucidate the molecular mechanisms.Our previous study showed that 12-day treatment of hHSCs with WOL could upregulate the expression of hypoxia-inducible factor 1-a (HIF1A) (Kondo et al., 2021), a key modulator of the transcriptional response to hypoxia and has been reported to regulate erythropoietin (Epo) production, which plays an important role in erythropoiesis. So, we hypothesize that WOL treatment would activate HIF1A in anemia-induced mice model, thus regulating Epo production to accelerate erythroid maturation. HIF2A is also reported to regulate Epo in kidney and Fpn in intestine to regulate erythropoiesis (Andrew J., et al., 2019).Therefore, we will investigate Epo expression in kidney, Hif1a and Hif2a expressions in kidney and intestine, and transferrin receptor (Tfrc, erythroid marker) in bone marrow. All gene expressions will be confirmed in protein levels also. [Display omitted] DisclosuresSuidasari: Nutrition Act Co. Ltd.: Current Employment. Yokozawa: Nutrition Act Co. Ltd.: Current Employment. Yamauchi: Nutrition Act Co. Ltd.: Current Employment.

The Protective Effect of Pistacia vera Pericarp on Kidney Function in Rats with Hemolytic Anemia

Background and objectives: Pistacia vera is known as a source of unique materials with therapeutic function such as antioxidant and nephron-protective activities. This study aimed to identify the biochemical and histopathological effects of Pistacia vera pericarp aqueous extract on the kidney in phenylhydrazine-induced anemia model in rats. Methods: Extraction of the P. vera pericarp was carried out by maceration technique. For animal study, the rats were studied in six groups and were exposed to phenylhydrazine for two days in the absence or presence of the extract. Renal changes were measured using biochemical and histopathological assays. The urine samples were collected in metabolic cages for total urine volume, creatinine, and 24-hour proteinuria measurement with the protein/creatinine ratio. Serum catalase, malondialdehyde and superoxide dismutase as oxidative stress markers were examined using ELISA test. Results: Phenylhydrazine induced kidney injuries evidenced by significant changes of urine, serum urea, creatinine levels, sodium, and potassium ions in comparison to the control group; however, the extract treatment significantly decreased kidney injuries. Administration of 80 mg/kg of the extract significantly reduced the creatinine and proteinuria in treated animals (p<0.05) but 160 mg/kg of extract helped the anemic animals to reduce protein and creatinine to normal levels. Conclusion: Twenty-four hours protein and creatinine can be used as markers of renal injuries in anemia and their regular measurement can be useful to find the risk of renal problems in anemia. These results revealed that P. vera pericarp administration may decrease renal injuries and dysfunction by reducing inflammation in the kidney.