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Investigation into the effects of antioxidant-rich extract of Tamarindus indica leaf on antioxidant enzyme activities, oxidative stress and gene expression profiles in HepG2 cells.

Razali N, Abdul Aziz A, Lim CY, Mat Junit S - PeerJ (2015)

Bottom Line: When the significantly regulated genes were analyzed using Ingenuity Pathway Analysis software, "Lipid Metabolism, Small Molecule Biochemistry, Hematological Disease" was the top biological network affected by the leaf extract, with a score of 36.The antioxidant-rich leaf extract of T. indica also altered the expression of proteins that are involved in the Coagulation System and the Intrinsic Prothrombin Activation Pathway (KNG1, SERPINE1, FGG), Superpathway of Cholesterol Biosynthesis (MVK), Immune protection/antimicrobial response (IFNGR1, LEAP2, ANXA3 and MX1) and Xenobiotic Metabolism Signaling (ALDH6A1, ADH6).In conclusion, the antioxidant-rich leaf extract of T. indica inhibited lipid peroxidation and ROS production, enhanced antioxidant enzyme activities and significantly regulated the expression of genes and proteins involved with consequential impact on the coagulation system, cholesterol biosynthesis, xenobiotic metabolism signaling and antimicrobial response.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.

ABSTRACT
The leaf extract of Tamarindus indica L. (T. indica) had been reported to possess high phenolic content and showed high antioxidant activities. In this study, the effects of the antioxidant-rich leaf extract of the T. indica on lipid peroxidation, antioxidant enzyme activities, H2O2-induced ROS production and gene expression patterns were investigated in liver HepG2 cells. Lipid peroxidation and ROS production were inhibited and the activity of antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase was enhanced when the cells were treated with the antioxidant-rich leaf extract. cDNA microarray analysis revealed that 207 genes were significantly regulated by at least 1.5-fold (p < 0.05) in cells treated with the antioxidant-rich leaf extract. The expression of KNG1, SERPINC1, SERPIND1, SERPINE1, FGG, FGA, MVK, DHCR24, CYP24A1, ALDH6A1, EPHX1 and LEAP2 were amongst the highly regulated. When the significantly regulated genes were analyzed using Ingenuity Pathway Analysis software, "Lipid Metabolism, Small Molecule Biochemistry, Hematological Disease" was the top biological network affected by the leaf extract, with a score of 36. The top predicted canonical pathway affected by the leaf extract was the coagulation system (P < 2.80 × 10(-6)) followed by the superpathway of cholesterol biosynthesis (P < 2.17 × 10(-4)), intrinsic prothrombin pathway (P < 2.92 × 10(-4)), Immune Protection/Antimicrobial Response (P < 2.28 × 10(-3)) and xenobiotic metabolism signaling (P < 2.41 × 10(-3)). The antioxidant-rich leaf extract of T. indica also altered the expression of proteins that are involved in the Coagulation System and the Intrinsic Prothrombin Activation Pathway (KNG1, SERPINE1, FGG), Superpathway of Cholesterol Biosynthesis (MVK), Immune protection/antimicrobial response (IFNGR1, LEAP2, ANXA3 and MX1) and Xenobiotic Metabolism Signaling (ALDH6A1, ADH6). In conclusion, the antioxidant-rich leaf extract of T. indica inhibited lipid peroxidation and ROS production, enhanced antioxidant enzyme activities and significantly regulated the expression of genes and proteins involved with consequential impact on the coagulation system, cholesterol biosynthesis, xenobiotic metabolism signaling and antimicrobial response.

No MeSH data available.


Related in: MedlinePlus

PCA plot.(A) A Principal Component Analysis (PCA) plot generated using Partek software of HepG2 cells grown in the presence or absence of the methanol leaf extract of T. indica. (Data are clustered based on three biological replicates (n = 3) of control and treated samples. Arrays for the untreated group are in blue and those for the treated group are in green. Each ball represents a sample. (B) Hierarchical clustering of highly significantly expressed genes that are associated to different pathways generated by Genesis software. The gene expression was regulated in response to the treatment with leaf extract on HepG2 cells. The regulation pattern was differentiated with 2 colours; green for down regulation and red for up regulation. The clustering was generated using Genesis software.
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fig-3: PCA plot.(A) A Principal Component Analysis (PCA) plot generated using Partek software of HepG2 cells grown in the presence or absence of the methanol leaf extract of T. indica. (Data are clustered based on three biological replicates (n = 3) of control and treated samples. Arrays for the untreated group are in blue and those for the treated group are in green. Each ball represents a sample. (B) Hierarchical clustering of highly significantly expressed genes that are associated to different pathways generated by Genesis software. The gene expression was regulated in response to the treatment with leaf extract on HepG2 cells. The regulation pattern was differentiated with 2 colours; green for down regulation and red for up regulation. The clustering was generated using Genesis software.

Mentions: Microarray data were filtered using Partek Genomics Suite software according to P value less than 0.05 and with fold change difference of equal to or greater than 1.5. Principle Component Analysis (PCA) plot was then generated using the same software to indicate the reproducibility of the microarray data (Fig. 3A). Biological replicates of control (n = 3) and leaf-treated HepG2 cells (n = 3) are shown in blue and green, respectively. From the plot, it is clear that the control samples were grouped separately from the treated samples. Hierarchical clustering in Fig. 3B shows the expression pattern of selected significantly regulated genes in each replicate of the untreated and T. indica leaf-treated cells. Green and red boxes indicate down- and up-regulated genes, respectively. A total of 207 genes were significantly regulated in leaf-treated HepG2 cells. The complete list of significantly regulated genes is attached in Table S2. Based on GO analyses, the list of highly significant genes categorized under their biological functions with details of the GenBank accession number, name of the gene and its respective protein, and fold change difference between treated and untreated cells are presented in Table 1. Amongst the significant regulated genes, the expression of CYP24A1 and SHBG showed the highest fold change difference of −4.19 and 4.11, respectively. Other significantly regulated genes include KNG1, SERPINC1, SERPIND1, SERPINE1, FGG, FGA, DHCR24, LEAP2, IFNGR1, ANXA3, MX1, ADH6 and ALDH6A1 (Table 1). Similar pattern of expression was generated when selected genes namely AREG, CYP24A1, ANXA3, FGG, FGA, LEAP2, SERPINE1, MVK, DHCR24, IFNGR1, ADH6 and ALDH6A1 were quantitated relative to that of GADPH, using qRT-PCR (Fig. 4).


Investigation into the effects of antioxidant-rich extract of Tamarindus indica leaf on antioxidant enzyme activities, oxidative stress and gene expression profiles in HepG2 cells.

Razali N, Abdul Aziz A, Lim CY, Mat Junit S - PeerJ (2015)

PCA plot.(A) A Principal Component Analysis (PCA) plot generated using Partek software of HepG2 cells grown in the presence or absence of the methanol leaf extract of T. indica. (Data are clustered based on three biological replicates (n = 3) of control and treated samples. Arrays for the untreated group are in blue and those for the treated group are in green. Each ball represents a sample. (B) Hierarchical clustering of highly significantly expressed genes that are associated to different pathways generated by Genesis software. The gene expression was regulated in response to the treatment with leaf extract on HepG2 cells. The regulation pattern was differentiated with 2 colours; green for down regulation and red for up regulation. The clustering was generated using Genesis software.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4636403&req=5

fig-3: PCA plot.(A) A Principal Component Analysis (PCA) plot generated using Partek software of HepG2 cells grown in the presence or absence of the methanol leaf extract of T. indica. (Data are clustered based on three biological replicates (n = 3) of control and treated samples. Arrays for the untreated group are in blue and those for the treated group are in green. Each ball represents a sample. (B) Hierarchical clustering of highly significantly expressed genes that are associated to different pathways generated by Genesis software. The gene expression was regulated in response to the treatment with leaf extract on HepG2 cells. The regulation pattern was differentiated with 2 colours; green for down regulation and red for up regulation. The clustering was generated using Genesis software.
Mentions: Microarray data were filtered using Partek Genomics Suite software according to P value less than 0.05 and with fold change difference of equal to or greater than 1.5. Principle Component Analysis (PCA) plot was then generated using the same software to indicate the reproducibility of the microarray data (Fig. 3A). Biological replicates of control (n = 3) and leaf-treated HepG2 cells (n = 3) are shown in blue and green, respectively. From the plot, it is clear that the control samples were grouped separately from the treated samples. Hierarchical clustering in Fig. 3B shows the expression pattern of selected significantly regulated genes in each replicate of the untreated and T. indica leaf-treated cells. Green and red boxes indicate down- and up-regulated genes, respectively. A total of 207 genes were significantly regulated in leaf-treated HepG2 cells. The complete list of significantly regulated genes is attached in Table S2. Based on GO analyses, the list of highly significant genes categorized under their biological functions with details of the GenBank accession number, name of the gene and its respective protein, and fold change difference between treated and untreated cells are presented in Table 1. Amongst the significant regulated genes, the expression of CYP24A1 and SHBG showed the highest fold change difference of −4.19 and 4.11, respectively. Other significantly regulated genes include KNG1, SERPINC1, SERPIND1, SERPINE1, FGG, FGA, DHCR24, LEAP2, IFNGR1, ANXA3, MX1, ADH6 and ALDH6A1 (Table 1). Similar pattern of expression was generated when selected genes namely AREG, CYP24A1, ANXA3, FGG, FGA, LEAP2, SERPINE1, MVK, DHCR24, IFNGR1, ADH6 and ALDH6A1 were quantitated relative to that of GADPH, using qRT-PCR (Fig. 4).

Bottom Line: When the significantly regulated genes were analyzed using Ingenuity Pathway Analysis software, "Lipid Metabolism, Small Molecule Biochemistry, Hematological Disease" was the top biological network affected by the leaf extract, with a score of 36.The antioxidant-rich leaf extract of T. indica also altered the expression of proteins that are involved in the Coagulation System and the Intrinsic Prothrombin Activation Pathway (KNG1, SERPINE1, FGG), Superpathway of Cholesterol Biosynthesis (MVK), Immune protection/antimicrobial response (IFNGR1, LEAP2, ANXA3 and MX1) and Xenobiotic Metabolism Signaling (ALDH6A1, ADH6).In conclusion, the antioxidant-rich leaf extract of T. indica inhibited lipid peroxidation and ROS production, enhanced antioxidant enzyme activities and significantly regulated the expression of genes and proteins involved with consequential impact on the coagulation system, cholesterol biosynthesis, xenobiotic metabolism signaling and antimicrobial response.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.

ABSTRACT
The leaf extract of Tamarindus indica L. (T. indica) had been reported to possess high phenolic content and showed high antioxidant activities. In this study, the effects of the antioxidant-rich leaf extract of the T. indica on lipid peroxidation, antioxidant enzyme activities, H2O2-induced ROS production and gene expression patterns were investigated in liver HepG2 cells. Lipid peroxidation and ROS production were inhibited and the activity of antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase was enhanced when the cells were treated with the antioxidant-rich leaf extract. cDNA microarray analysis revealed that 207 genes were significantly regulated by at least 1.5-fold (p < 0.05) in cells treated with the antioxidant-rich leaf extract. The expression of KNG1, SERPINC1, SERPIND1, SERPINE1, FGG, FGA, MVK, DHCR24, CYP24A1, ALDH6A1, EPHX1 and LEAP2 were amongst the highly regulated. When the significantly regulated genes were analyzed using Ingenuity Pathway Analysis software, "Lipid Metabolism, Small Molecule Biochemistry, Hematological Disease" was the top biological network affected by the leaf extract, with a score of 36. The top predicted canonical pathway affected by the leaf extract was the coagulation system (P < 2.80 × 10(-6)) followed by the superpathway of cholesterol biosynthesis (P < 2.17 × 10(-4)), intrinsic prothrombin pathway (P < 2.92 × 10(-4)), Immune Protection/Antimicrobial Response (P < 2.28 × 10(-3)) and xenobiotic metabolism signaling (P < 2.41 × 10(-3)). The antioxidant-rich leaf extract of T. indica also altered the expression of proteins that are involved in the Coagulation System and the Intrinsic Prothrombin Activation Pathway (KNG1, SERPINE1, FGG), Superpathway of Cholesterol Biosynthesis (MVK), Immune protection/antimicrobial response (IFNGR1, LEAP2, ANXA3 and MX1) and Xenobiotic Metabolism Signaling (ALDH6A1, ADH6). In conclusion, the antioxidant-rich leaf extract of T. indica inhibited lipid peroxidation and ROS production, enhanced antioxidant enzyme activities and significantly regulated the expression of genes and proteins involved with consequential impact on the coagulation system, cholesterol biosynthesis, xenobiotic metabolism signaling and antimicrobial response.

No MeSH data available.


Related in: MedlinePlus