Asian Pacific Journal of Tropical Biomedicine
Issue 12,2018 Table of Contents
2018(12):565-570.
DOI: 10.4103/2221-1691.248092
Abstract:
Objective: To determine the biochemical and acute phase proteins changes in sheep experimentally infected with Anaplasma ovis (A. ovis). Methods: One Iranian sheep naturally infected with A. ovis (parasitemia 0.02%) but with no other blood parasites based on blood smear and polymerase chain reaction methods was selected as donor, and it was splenectomized to induce high level of parasitemia. Then, three weeks after splenectomy when parasitemia was 6%, donor’s blood was intravenously administered to each recipient animal. Five 5-6 months old Iranian male sheep without any blood parasites were selected as recipient animals. The percent of parasites, packed cell volume, serum biochemical parameters (urea, creatinine, bilirubin, aspartate aminotransferase activity, cholesterol, total protein, albumin, globulin, Fe), acute phase proteins (haptoglobin, total iron binding capacity, fibrinogen), were evaluated in sheep before and after being experimentally infected with A. ovis (until day 38). In addition, body weights of sheep were measured on days 0, 20 and 38. Results: In recipient sheep, microscopic examination of erythrocytes revealed a significant rise of parasitemia on days 12 and 15. The lowest level of packed cell volume in sheep was seen on day 15 post infection. A significant rise existed in mean urea and bilirubin (total, direct and indirect) on days 15 and 20. The increase of indirect bilirubin level was higher than direct bilirubin. Furthermore, serum Fe significantly increased on days 20 and 23. The mean total protein concentration significantly increased on day 38. A significant increase was found in the serum globulin concentration from days 20 and 27 to 38. Maximum values of haptoglobin were observed on days 27 and 30. Moreover, aspartate aminotransferase activity (from days 20-30) and cholesterol concentration (on day 20) significantly decreased. However, no significant changes were found in other parameters. Conclusions: Experimental ovine anaplasmosis caused by A. ovis could be associated with some changes in measured parameters, which presumably could be helpful for evaluation on staging of disease.
Objective: To determine the biochemical and acute phase proteins changes in sheep experimentally infected with Anaplasma ovis (A. ovis). Methods: One Iranian sheep naturally infected with A. ovis (parasitemia 0.02%) but with no other blood parasites based on blood smear and polymerase chain reaction methods was selected as donor, and it was splenectomized to induce high level of parasitemia. Then, three weeks after splenectomy when parasitemia was 6%, donor’s blood was intravenously administered to each recipient animal. Five 5-6 months old Iranian male sheep without any blood parasites were selected as recipient animals. The percent of parasites, packed cell volume, serum biochemical parameters (urea, creatinine, bilirubin, aspartate aminotransferase activity, cholesterol, total protein, albumin, globulin, Fe), acute phase proteins (haptoglobin, total iron binding capacity, fibrinogen), were evaluated in sheep before and after being experimentally infected with A. ovis (until day 38). In addition, body weights of sheep were measured on days 0, 20 and 38. Results: In recipient sheep, microscopic examination of erythrocytes revealed a significant rise of parasitemia on days 12 and 15. The lowest level of packed cell volume in sheep was seen on day 15 post infection. A significant rise existed in mean urea and bilirubin (total, direct and indirect) on days 15 and 20. The increase of indirect bilirubin level was higher than direct bilirubin. Furthermore, serum Fe significantly increased on days 20 and 23. The mean total protein concentration significantly increased on day 38. A significant increase was found in the serum globulin concentration from days 20 and 27 to 38. Maximum values of haptoglobin were observed on days 27 and 30. Moreover, aspartate aminotransferase activity (from days 20-30) and cholesterol concentration (on day 20) significantly decreased. However, no significant changes were found in other parameters. Conclusions: Experimental ovine anaplasmosis caused by A. ovis could be associated with some changes in measured parameters, which presumably could be helpful for evaluation on staging of disease.
2018(12):571-579.
DOI: 10.4103/2221-1691.248093
Abstract:
Objective: To investigate the impact of the extracts of Gac fruit parts (peel, pulp, seed, and aril) on the cell viability and angiogenesis markers of human retinal pigment epithelial (ARPE19) cells under high glucose conditions. Methods: The effect of the extracts of Gac fruit peel, pulp, seed and aril on the ARPE-19 cells was determined using MTT viability assay, Trypan blue dye and morphological changes were observed using light microscopy. Enzyme-linked immunosorbent-based assay was performed to evaluate the effect of Gac fruit parts on the reactive oxygen species (ROS), vascular endothelial growth factor (VEGF) and pigmented epithelium-derived factor (PEDF) secretions. Results: High glucose (HG) at 30 mmol/L increased ARPE-19 cell viability and ROS and VEGF secretions. While, the exposure of ARPE19 cells in high glucose condition to Gac fruit extracts led to inhibition of cell viability, induced morphological changes, decreased ROS and VEGF secretions, and increased PEDF level. Gac pulp, seed, and aril at 1 000 μg/mL showed significant inhibition activities [(7.5 ± 5.1)%, (2.7 ± 0.5)%, (3.2 ± 1.1)%, respectively] against HG-induced ARPE-19 cell viability. The findings also demonstrated that Gac aril at 250 μg/mL significantly decreased ROS and VEGF levels [(40.6 ± 3.3) pg/mL, (107.4 ± 48.3) pg/mL, respectively] compared to ROS [(71.7 ± 2.9) pg/ mL] and VEGF [(606.9 ± 81.1) pg/mL] in HG untreated cells. Moreover, 250 μg/mL of Gac peel dramatically increased PEDF level [(18.2 ± 0.3) ng/mL] compared to that in HG untreated cells [(0.48 ± 0.39) ng/mL]. Conclusions: This study indicates that the extracts of Gac peel, pulp, seed and aril reduced cell viability, minimized ROS generations and showed angiogenic activities. Therefore, our findings open new insights into the potentiality of Gac fruit against HG-related diabetic retinopathy disease.
Objective: To investigate the impact of the extracts of Gac fruit parts (peel, pulp, seed, and aril) on the cell viability and angiogenesis markers of human retinal pigment epithelial (ARPE19) cells under high glucose conditions. Methods: The effect of the extracts of Gac fruit peel, pulp, seed and aril on the ARPE-19 cells was determined using MTT viability assay, Trypan blue dye and morphological changes were observed using light microscopy. Enzyme-linked immunosorbent-based assay was performed to evaluate the effect of Gac fruit parts on the reactive oxygen species (ROS), vascular endothelial growth factor (VEGF) and pigmented epithelium-derived factor (PEDF) secretions. Results: High glucose (HG) at 30 mmol/L increased ARPE-19 cell viability and ROS and VEGF secretions. While, the exposure of ARPE19 cells in high glucose condition to Gac fruit extracts led to inhibition of cell viability, induced morphological changes, decreased ROS and VEGF secretions, and increased PEDF level. Gac pulp, seed, and aril at 1 000 μg/mL showed significant inhibition activities [(7.5 ± 5.1)%, (2.7 ± 0.5)%, (3.2 ± 1.1)%, respectively] against HG-induced ARPE-19 cell viability. The findings also demonstrated that Gac aril at 250 μg/mL significantly decreased ROS and VEGF levels [(40.6 ± 3.3) pg/mL, (107.4 ± 48.3) pg/mL, respectively] compared to ROS [(71.7 ± 2.9) pg/ mL] and VEGF [(606.9 ± 81.1) pg/mL] in HG untreated cells. Moreover, 250 μg/mL of Gac peel dramatically increased PEDF level [(18.2 ± 0.3) ng/mL] compared to that in HG untreated cells [(0.48 ± 0.39) ng/mL]. Conclusions: This study indicates that the extracts of Gac peel, pulp, seed and aril reduced cell viability, minimized ROS generations and showed angiogenic activities. Therefore, our findings open new insights into the potentiality of Gac fruit against HG-related diabetic retinopathy disease.
2018(12):580-585.
DOI: 10.4103/2221-1691.248094
Abstract:
Objective: To investigate anti-hemolytic, antibacterial and anti-cancer activities of leaf and stem extracts from Polygonum odoratum. Methods: Leaves and stems of Polygonum odoratum were extracted using methanol and their anti-hemolytic activity was assessed using 2, 2′-Azobis (2-methylpropionamidine) dihydrochloride which is known to generate free radical damage on cell membranes of red blood cells. This damage, represented by hemolysis, was measured using spectrophotometry. Antibacterial activity was tested by using a broth microdilution method to find minimal inhibitory concentrations against eight bacterial strains. Anti-cancer activity of the extracts was evaluated against a human promyelocytic leukemic cell line (HL60) by using MTT assay for cell viability and flow cytometry for apoptosis induction and cell cycle analysis. Results: Both leaf and stem extracts have anti-hemolytic activity. The results showed a significantly increased percentage of inhibition in a concentration-dependent manner. Interestingly, the leaf extract showed anti-hemolytic activity to a greater extent than the stem extract. Antibacterial activity of the extracts, as indicated by their minimal inhibitory concentration, using 12.5, 50, 25, 25 μg/mL, was measured against Staphylococcus epidermidis, Enterococcus faecium, Enterococcus faecalis and Staphylococcus aureus. The leaf extracts also exhibited anti-cancer activity, demonstrated by significantly decreased cell viability of human promyelocytic cells (HL-60), with an IC50 of (350.00+1.85) μg/mL for 48 h and (38.00+0.92) μg/mL for 72 h. Additionally, HL-60 became apoptotic and accumulated in G1-phase after 48 hours of treatment. Conclusions: The extracts of Polygonum odoratum exhibit potential anti hemolytic activity. They also have antibacterial activity by inhibiting growth of Gram-positive bacteria. The leaf extract shows anti-cancer activity against HL-60 to a greater extent than the stem extract, causing decreased viability, increased G1-phase accumulation and apoptosis induction.
Objective: To investigate anti-hemolytic, antibacterial and anti-cancer activities of leaf and stem extracts from Polygonum odoratum. Methods: Leaves and stems of Polygonum odoratum were extracted using methanol and their anti-hemolytic activity was assessed using 2, 2′-Azobis (2-methylpropionamidine) dihydrochloride which is known to generate free radical damage on cell membranes of red blood cells. This damage, represented by hemolysis, was measured using spectrophotometry. Antibacterial activity was tested by using a broth microdilution method to find minimal inhibitory concentrations against eight bacterial strains. Anti-cancer activity of the extracts was evaluated against a human promyelocytic leukemic cell line (HL60) by using MTT assay for cell viability and flow cytometry for apoptosis induction and cell cycle analysis. Results: Both leaf and stem extracts have anti-hemolytic activity. The results showed a significantly increased percentage of inhibition in a concentration-dependent manner. Interestingly, the leaf extract showed anti-hemolytic activity to a greater extent than the stem extract. Antibacterial activity of the extracts, as indicated by their minimal inhibitory concentration, using 12.5, 50, 25, 25 μg/mL, was measured against Staphylococcus epidermidis, Enterococcus faecium, Enterococcus faecalis and Staphylococcus aureus. The leaf extracts also exhibited anti-cancer activity, demonstrated by significantly decreased cell viability of human promyelocytic cells (HL-60), with an IC50 of (350.00+1.85) μg/mL for 48 h and (38.00+0.92) μg/mL for 72 h. Additionally, HL-60 became apoptotic and accumulated in G1-phase after 48 hours of treatment. Conclusions: The extracts of Polygonum odoratum exhibit potential anti hemolytic activity. They also have antibacterial activity by inhibiting growth of Gram-positive bacteria. The leaf extract shows anti-cancer activity against HL-60 to a greater extent than the stem extract, causing decreased viability, increased G1-phase accumulation and apoptosis induction.
2018(12):586-592.
DOI: 10.4103/2221-1691.248095
Abstract:
Objective: To determine the anti-cancer effect of aronia leaf extract on SK-Hep1 cells using migration, metallo metrix proteinase-2/-9 (MMP-2/-9) and MT-1 MMP expression and to evaluate the anti-inflammatory activities of the leaf extract. Methods: The effect of aronia leaf extract on cancer prevention was investigated. SK-Hep1 human liver cancer cell line was treated with aronia leaf extract at various concentractions. MTT assay was used to measure cancer cell growth inhibition, and wound migration assay was used for metastasis determination. The expression of MMP-2/-9 was measured at the protein level using zymography and the expression of MMP-2/-9 and MT-1 MMP was examined at the gene level by RT-PCR. Raw 264.7 macrophage cells were stimulated with lipopolysaccharides to induce inflammation, and then the inhibition of inflammation was evaluated by treatment of aronia leaf extract. Expressions of interleukin-6, tumor factor-a, and nitric oxide (NO) were also determined. Results: SK-Hep1 cell growth was inhibited in proportion to the concentration of aronia leaf extract. In migration assay, aronia leaf extract showed 61.3%-96.3% wound size inhibtion after treating 50-200 μg/mL of aronia leaf extract for 24 h. At the protein level, the expression of MMP-2 and MMP-9 decreased as the concentration of aronia leaf extract treated with SK-Hep1 cells increased. In addition, the same pattern as in the protein was also observed in the mRNA levels. The expressions of MMP-2 and MMP-9 protein were inhibited by 92.2% and 53.8%, respectively after treatment with 200 μg/mL aronia leaf extract. In addition, Raw 264.7 cells treated with aronia leaf extract did not affect cell survival. There was dose dependent inhibition of interleukine-6, tumor necrosis factor-a and nitric oxide after treating aronia leaf extract in lipopolysaccharides-treated Raw 264.7 cell. Conclusions: The results show that aronia leaf has anticancer and and antimetastatic properties in SK-Hep1 and Raw 264.7 cells.
Objective: To determine the anti-cancer effect of aronia leaf extract on SK-Hep1 cells using migration, metallo metrix proteinase-2/-9 (MMP-2/-9) and MT-1 MMP expression and to evaluate the anti-inflammatory activities of the leaf extract. Methods: The effect of aronia leaf extract on cancer prevention was investigated. SK-Hep1 human liver cancer cell line was treated with aronia leaf extract at various concentractions. MTT assay was used to measure cancer cell growth inhibition, and wound migration assay was used for metastasis determination. The expression of MMP-2/-9 was measured at the protein level using zymography and the expression of MMP-2/-9 and MT-1 MMP was examined at the gene level by RT-PCR. Raw 264.7 macrophage cells were stimulated with lipopolysaccharides to induce inflammation, and then the inhibition of inflammation was evaluated by treatment of aronia leaf extract. Expressions of interleukin-6, tumor factor-a, and nitric oxide (NO) were also determined. Results: SK-Hep1 cell growth was inhibited in proportion to the concentration of aronia leaf extract. In migration assay, aronia leaf extract showed 61.3%-96.3% wound size inhibtion after treating 50-200 μg/mL of aronia leaf extract for 24 h. At the protein level, the expression of MMP-2 and MMP-9 decreased as the concentration of aronia leaf extract treated with SK-Hep1 cells increased. In addition, the same pattern as in the protein was also observed in the mRNA levels. The expressions of MMP-2 and MMP-9 protein were inhibited by 92.2% and 53.8%, respectively after treatment with 200 μg/mL aronia leaf extract. In addition, Raw 264.7 cells treated with aronia leaf extract did not affect cell survival. There was dose dependent inhibition of interleukine-6, tumor necrosis factor-a and nitric oxide after treating aronia leaf extract in lipopolysaccharides-treated Raw 264.7 cell. Conclusions: The results show that aronia leaf has anticancer and and antimetastatic properties in SK-Hep1 and Raw 264.7 cells.
2018(12):593-597.
DOI: 10.4103/2221-1691.248096
Abstract:
Objective: To investigate antidiabetic effects of Tetracarpidium conophorum seed (TECOSE) against biomarkers of diabetes-induced nephropathy in rats. Methods: Powdered seed (500 g) of TECOSE was extracted with 5 L of 100% methanol for 72 h followed by concentration of filtrate. Diabetes was induced in rats with 75 mg/kg body weight intraperitoneal streptozotocin. The rats were randomly divided into five groups (n=5 in each group) namely group A- normal control, group B- diabetic control, groups C, D and E were diabetic rats treated with 500 mg/kg body weight TECOSE orally, 7 mg/kg body weight metformin orally and subcutaneous 0.3 IU/kg body weight HumulinR, respectively. All rats were treated once daily for 2 weeks. Blood samples and urine were collected for biochemical estimations. Kidney was harvested for histomorphological studies. Results: TECOSE (500 mg/kg body weight) significantly (P<0.05) reduced blood sugar levels as effective as metformin and insulin. The plant extract also significantly (P<0.05) reduced levels of serum urea, creatinine and uric acid, proteinuria, ketonuria, hematuria and glycosuria while it significantly (P<0.05) increased glomerular filtration rate. Histomorphological study of the kidney of untreated diabetic rats showed features suggestive of glomerulosclerosis and tubular necrosis while that of treatments with TECOSE extract, metformin and insulin showed near normal histoarchitectures. Conclusions: Streptozotocin at 75 mg/kg body weight induces diabetic nephropathy in rats and TECOSE possesses potentials to prevent diabetic renal damage.
Objective: To investigate antidiabetic effects of Tetracarpidium conophorum seed (TECOSE) against biomarkers of diabetes-induced nephropathy in rats. Methods: Powdered seed (500 g) of TECOSE was extracted with 5 L of 100% methanol for 72 h followed by concentration of filtrate. Diabetes was induced in rats with 75 mg/kg body weight intraperitoneal streptozotocin. The rats were randomly divided into five groups (n=5 in each group) namely group A- normal control, group B- diabetic control, groups C, D and E were diabetic rats treated with 500 mg/kg body weight TECOSE orally, 7 mg/kg body weight metformin orally and subcutaneous 0.3 IU/kg body weight HumulinR, respectively. All rats were treated once daily for 2 weeks. Blood samples and urine were collected for biochemical estimations. Kidney was harvested for histomorphological studies. Results: TECOSE (500 mg/kg body weight) significantly (P<0.05) reduced blood sugar levels as effective as metformin and insulin. The plant extract also significantly (P<0.05) reduced levels of serum urea, creatinine and uric acid, proteinuria, ketonuria, hematuria and glycosuria while it significantly (P<0.05) increased glomerular filtration rate. Histomorphological study of the kidney of untreated diabetic rats showed features suggestive of glomerulosclerosis and tubular necrosis while that of treatments with TECOSE extract, metformin and insulin showed near normal histoarchitectures. Conclusions: Streptozotocin at 75 mg/kg body weight induces diabetic nephropathy in rats and TECOSE possesses potentials to prevent diabetic renal damage.
2018(12):598-603.
DOI: 10.4103/2221-1691.248097
Abstract:
Objective: To investigate the cytotoxicity and anti-cancer effects of hydro-alcoholic extract of pistachio pericarp on hepatocellular carcinoma cells (HepG2) and mouse fibroblast L929 cells as normal and control group cell. Methods: MTT assay was performed to investigate the cytotoxicity effects of the extract at 0-4 000 μg/mL on the cells after 24 and 48 h. The expressions of some genes involved in apoptosis including Bax, Bcl-2 and P53 were investigated by real time PCR. Results: Our results showed that after 24 and 48 hours of treatment of cells with this extract, the viability of HepG2 and L929 cells was reduced. Therefore, this extract had the cytotoxicity effect on both cells. The IC50 concentration of extract for HepG2 cells after 24 and 48 hours of treatment was 1 500 and 1 000 μg/mL and for L929 cells was 2 000 and 1 500 μg/mL, respectively. The expressions of Bax and P53 genes were up-regulated after treatment in the HepG2 and L929 cells and the expression of Bcl-2 gene was down-regulated after treatment of extract in HepG2 cell. Conclusions: According to the results of MTT assay and real time PCR, this extract can be considered as a potential candidate for use in the production of anti-cancer drugs for the treatment of patients with liver cancer in future.
Objective: To investigate the cytotoxicity and anti-cancer effects of hydro-alcoholic extract of pistachio pericarp on hepatocellular carcinoma cells (HepG2) and mouse fibroblast L929 cells as normal and control group cell. Methods: MTT assay was performed to investigate the cytotoxicity effects of the extract at 0-4 000 μg/mL on the cells after 24 and 48 h. The expressions of some genes involved in apoptosis including Bax, Bcl-2 and P53 were investigated by real time PCR. Results: Our results showed that after 24 and 48 hours of treatment of cells with this extract, the viability of HepG2 and L929 cells was reduced. Therefore, this extract had the cytotoxicity effect on both cells. The IC50 concentration of extract for HepG2 cells after 24 and 48 hours of treatment was 1 500 and 1 000 μg/mL and for L929 cells was 2 000 and 1 500 μg/mL, respectively. The expressions of Bax and P53 genes were up-regulated after treatment in the HepG2 and L929 cells and the expression of Bcl-2 gene was down-regulated after treatment of extract in HepG2 cell. Conclusions: According to the results of MTT assay and real time PCR, this extract can be considered as a potential candidate for use in the production of anti-cancer drugs for the treatment of patients with liver cancer in future.
Jarinyaporn Naowaboot
,
Pritsana Piyabhan
,
Pholawat Tingpej
,
Narongsuk Munkong
,
Wason Parklak
,
Patchareewan Pannangpetch
2018(12):604-608.
DOI: 10.4103/2221-1691.248098
Abstract:
Objective: To evaluate the insulin sensitivity action of ferulic acid (FA) in skeletal muscle and hypothalamus of high-fat diet (HFD)-induced obese mice. Methods: Obese mouse model was induced by HFD (45 kcal% lard fat) for 16 weeks. After 8 weeks of HFD feeding, these obese mice were orally treated with FA at doses of 25 and 50 mg/kg/day for 8 weeks. At the end of all treatments, the epididymal fat, pancreas, skeletal muscle and hypothalamus were removed for biochemical parameter and protein expression examinations. Results: FA treatment significantly decreased leptin level in fat tissue and insulin level in pancreas (P< 0.05). Interestingly, obese mice treated with FA increased the protein expressions of insulin receptor substrate-1, phosphatidylinositol 3-kinase, and phosphorylated-protein kinase B in both muscle and brain (P< 0.05). The phosphorylations of adenosine monophosphate-activated protein kinase and acetyl-CoA carboxylase in muscle, and leptin receptor protein in hypothalamus were also increased (P< 0.05). The pancreatic islets histology showed smaller size in obese mice treated with FA compared to untreated obese mice. Conclusions: These findings indicate the beneficial effect of FA in improving insulin resistance in HFD-induced obese mice. These effects are probably mediated via modulating the insulin receptor substrate/phosphatidylinositol 3-kinase/protein kinase B or adenosine monophosphate-activated protein kinase pathways.
Objective: To evaluate the insulin sensitivity action of ferulic acid (FA) in skeletal muscle and hypothalamus of high-fat diet (HFD)-induced obese mice. Methods: Obese mouse model was induced by HFD (45 kcal% lard fat) for 16 weeks. After 8 weeks of HFD feeding, these obese mice were orally treated with FA at doses of 25 and 50 mg/kg/day for 8 weeks. At the end of all treatments, the epididymal fat, pancreas, skeletal muscle and hypothalamus were removed for biochemical parameter and protein expression examinations. Results: FA treatment significantly decreased leptin level in fat tissue and insulin level in pancreas (P< 0.05). Interestingly, obese mice treated with FA increased the protein expressions of insulin receptor substrate-1, phosphatidylinositol 3-kinase, and phosphorylated-protein kinase B in both muscle and brain (P< 0.05). The phosphorylations of adenosine monophosphate-activated protein kinase and acetyl-CoA carboxylase in muscle, and leptin receptor protein in hypothalamus were also increased (P< 0.05). The pancreatic islets histology showed smaller size in obese mice treated with FA compared to untreated obese mice. Conclusions: These findings indicate the beneficial effect of FA in improving insulin resistance in HFD-induced obese mice. These effects are probably mediated via modulating the insulin receptor substrate/phosphatidylinositol 3-kinase/protein kinase B or adenosine monophosphate-activated protein kinase pathways.
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