Decaffeinated Green Tea and Green Coffee Extract Attenuate Cardiac Perivascular Fibrosis in a Metabolic Syndrome Model by Decreasing Fibroblast Growth Factor 23 and Runt-related transcription factor 2 Expression

Metabolic syndrome (MetS) is a group of risk factors in the form of central obesity, insulin resistance, dyslipidemia, and hypertension, increasing oxidative stress. This pathological event leads to the development of cardiovascular disease, for instance, atherosclerosis. Besides modifiable risk factors, non-modifiable risk factors such as genetic factors also play a role in the formation of atherosclerosis in MetS conditions such as THOC5, AIF1, CNN1, and ACTA2. Recently, natural compound derivatives, such as epigallocatechin-3-gallate (EGCG), chlorogenic acid (CGA), and turmeric, have shown beneficial effects in MetS improvement. This study aimed to investigate the effect of green tea, green coffee, and turmeric extract on the expression of THOC5, AIF1, CNN1, and ACTA2 genes that contributed to atherosclerotic vasculopathy development in the MetS rat model. Twenty-five MetS rat models were grouped into 4 groups (n = 5): Standard control (SC), MetS (MetS), a combination of green tea, green coffee, and turmeric extract with treatment doses: 300/100/150 mg/BW(C1) and 400/200/250 mg/BW(C2) group. The THOC5, AIF1, CNN1, and ACTA2 expression were measured at the end of treatment periods. This study found that administering green tea, green coffee, and turmeric extract can lower the expression of THOC5, AIF1, CNN1, and ACTA2. The correlation test showed that there is a strong correlation between THOC5 and AIF1 gene expression, with positive value. In summary, the combined effects of green tea, green coffee, and curcumin extract show significant promise as a potential anti-atherosclerosis treatment by improve the THOC5 and AIF1, but not ACTA2 and CNN1 gene expression in the aortic tissue of metabolic syndrome model. HIGHLIGHTS Metabolic syndrome is a group of risk factors consist of central obesity, insulin resistance, dyslipidemia, and hypertension. Individuals with MetS have an increased risk of developing atherosclerotic cardiovascular disease. Besides modifiable risk factors, non-modifiable risk factors such as genetic factors also play a role in the formation of atherosclerosis in MetS conditions such as THOC5, AIF1, CNN1, and ACTA2. This study aimed to investigate the effect of green tea, green coffee, and turmeric extract on the expression of THOC5, AIF1, CNN1, and ACTA2 genes that contributed to atherosclerotic vasculopathy development in the MetS rat model. Administering green tea, green coffee, and turmeric extract can lower the expression of THOC5, AIF1, CNN1, and ACTA2. The correlation test showed that there is a strong correlation between THOC5 and AIF1 gene expression, with positive value. In summary, the combined effects of green tea, green coffee, and curcumin extract show significant promise as a potential anti-atherosclerosis treatment by improve the THOC5 and AIF1, but not ACTA2 and CNN1 gene expression in the aortic tissue of metabolic syndrome model. GRAPHICAL ABSTRACT

Background: Metabolic syndrome is a significant risk factor for cardiovascular diseases. Green tea and green coffee extracts, antioxidant and anti-inflammatory agents may participate in metabolic syndrome-induced cardiac fibrosis alleviation. However, the effect of combination of those extracts still needs exploration. Therefore, this study investigated the effect of green tea and decaffeinated light roasted green coffee extracts and their combination in metabolic syndrome-induced cardiac fibrosis rats. Methods: Metabolic syndrome rat model was i1nduced through high-fat high sucrose diets feeding for 8 weeks and injection of low dose streptozotocin at the 2nd week. The metabolic syndrome rats were divided into 4 experimental groups metabolic syndrome rats (MS); metabolic syndrome rats treated with 300 mg/ kg b.w green tea extract (GT); metabolic syndrome rats treated with 200 mg/ kg b.w decaffeinated light roasted green coffee extract (GC); metabolic syndrome rats treated with the combination of the two extracts (CE); and a normal control (NC) group was added. Angiotensin 2 level was analyzed by ELISA method. Gene expression of NF-κB, TNF-α, IL-6, Tgf-β1, Rac-1, and α-sma were analyzed by touchdown polymerase chain reaction methods. Results: Metabolic syndrome rats treated with green tea and decaffeinated light roasted green coffee significantly decreased angiotensin-2 serum level and cardiac inflammation and fibrosis gene expression level (NF-κB, TNF-α, IL-6, Tgf-β1, Rac-1, and α-sma). More significant alleviation was observed in the combination group. Conclusion: This study suggested that combination of green tea and decaffeinated light roasted green coffee extracts showed better improvement in metabolic syndrome-induced cardiac fibrosis rat model compared to that of single extract administration through inflammation inhibition

Background: Individually, green tea and green coffee have been extensively studied for mitigation of metabolic syndrome (MS) in both rats and humans; however, their combined effect requires further investigation. Thus, we compared the metabolic effect of combining green tea and decaffeinated light roasted green coffee on MS in rats. Methods: An MS animal model was constructed by feeding Sprague-Dawley rats with a high-fat-high-sucrose (HFHS) diet for eight weeks and a low dose of streptozotocin (STZ) injection at week 2. Rats fed with HFHS diets and injected with STZ successfully developed MS phenotypes, indicated by higher body weight, systolic blood pressure, plasma triglyceride level, plasma fasting blood glucose level, and lower plasma HDL-C level, compared to those fed with a normal chow diet. Subsequently, MS rats were continuously fed with HFHS and divided into four groups: MS rats, MS with 300 mg/bw.t green tea extract (GT), MS with 200 mg/bw.t green coffee extract (GC), and MS with combined green tea and green coffee extract (CM) for nine weeks. Results: Combining green tea and green coffee have synergistic effects on reducing plasma fasting blood glucose and triglyceride level. Inflammatory markers both in plasma and liver tissue robustly decreased in CM group rats. However, the reduction of systolic blood pressure was observed only in GT and CM groups. Moreover, all treatment resulted in an increase in plasma HDL-C level in MS rats. Conclusions: Our data highlighted that, in MS animal models, combined green tea and decaffeinated light roasted green coffee augment their several individual beneficial effects of improved metabolic parameters and modulated inflammatory genes.

Background: Insulin resistance has been independently associated with cardiac diseases. Free fatty acids are recently known to induce cardiac insulin resistance due to low-grade inflammation. Therefore, the improvement of free fatty acid levels can also improve cardiac insulin resistance. This study investigated the combination of green tea and decaffeinated-light roasted green coffee extract in the improvement of free fatty acid-induced cardiac insulin resistance by improving the adiponectin/FAS pathways. Methods: This study used 25 males Sprague-Dawley rats induced by a high-fat high sucrose diet and injection of low dose streptozotocin to make a metabolic syndrome (MS) rat model and standard chow as healthy control rats. The MS rats were treated with green tea (200 mg/ b. w.), decaffeinated-light roasted green coffee (300 mg/ b. w.), and the combination of both extracts in 9 weeks. Experimental groups in this study were divided into 5 groups: 1) MS (HFHS diet + STZ) group, 2) NC (normal chow) group, 3) GT (green tea extract) group, 4) GC (decaffeinated-light roasted green coffee extract), 5) CM (combination of both extracts) group. Adiponectin and HOMA-IR level was analysed using ELISA, and the gene expression of Adipo-R1, FAS, PI3K, PDK1, Akt, GLUT4 was measured by RT-PCR. Results: The combination of green tea and decaffeinated-light roasted green coffee showed synergistic effects in improving FFA levels. The adiponectin/FAS pathways was attenuated in the CM group. Moreover, the combination also showed improvement in cardiac insulin resistance markers such as IRS1/2, PI3K, PDK1, Akt, and GLUT4. Conclusions: The combination of green tea and decaffeinated-light roasted green coffee extract improved cardiac insulin resistance better than green tea and green coffee extract administration alone by reducing free fatty acids levels through adiponectin/FAS pathways modulation.

Background: Insulin resistance has been independently associated with cardiac diseases. A free fatty acid is recently known to induce cardiac insulin resistance due to low-grade inflammation. Therefore, the improvement of free fatty acid levels can also improve cardiac insulin resistance. This study investigated the combination of green tea and decaffeinated-light roasted green coffee extract in improvement of free fatty acid-induced cardiac insulin resistance by improving the adiponectin/FAS pathway. Methods: This study used 25 males Sprague-Dawley rats induced by a high-fat high sucrose diet and injection of low dose streptozotocin to make a metabolic syndrome (MS) rat model and standard chow as healthy control rats. The MS rats were treated with green tea (200 mg/ b. w.), decaffeinated-light roasted green coffee (300 mg/ b. w.), and the combination of both extracts in 9 weeks. Experimental groups in this study were divided into 5 groups: 1) MS (HFHS diet + STZ) group, 2) NC (normal chow) group, 3) GT (green tea extract) group, 4) GC (decaffeinated-light roasted green coffee extract), 5) CM (combination of both extracts) group. Adiponectin and HOMA-IR level was analysed using ELISA, and the gene expression of Adipo-R1, FAS, PI3K, PDK1, Akt, GLUT4 was measured by RT-PCR. Results: The combination of green tea and decaffeinated-light roasted green coffee showed synergistic effects in improving FFA levels. The adiponectin/FAS pathway was attenuated in the CM group. Moreover, the combination also showed improvement in cardiac insulin resistance markers such as IRS1/2, PI3K, PDK1, Akt, and GLUT4. Conclusions: The combination of green tea and decaffeinated-light roasted green coffee extract improved cardiac insulin resistance better than green tea and green coffee extract administration alone by reducing free fatty acids levels through adiponectin/FAS pathway modulation.

Effect of green coffee and green tea extract on metabolic syndrome. To explore green coffee and green tea extract combination effect on metabolic profile and blood pressure improvement through adenosine monophosphate-activated protein kinase (AMPK) and Peroxisome Proliferator-Activated Receptor α (PPARα) gene expression modulation. Experimental laboratory research with pre- and post-control group design. Twenty-five metabolic syndrome rats model were grouped into five groups (n = 5): standard control (normal), metabolic syndrome (SM), green coffee extract (GC), green tea extract (GT), and combination green coffee and green tea extract (CM). The extract was given during 9 weeks. Serum glucose, triglyceride, high-density lipoprotein, and systolic blood pressure level were analyzed before and after the extract administration. At the end of the study, PPAR-α and AMPK-α2 gene were analyzed. Independent t-test. CM group had significantly higher PPAR-α, and AMPK-α2 gene expression compared to those of SM, GC, and GT group. Green coffee and green tea extract combination administration improved metabolic profile and blood pressure on metabolic syndrome through affecting PPAR-α and AMPK-α2 gene expression.

The present study evaluated the antioxidant effect of green and black tea (Camellia sinensis L.) extracts in uncured pork sausages. The total polyphenol content in the green tea extract (GTE) was significantly higher (p < 0.05) than the black tea extract (BTE). DPPH assay showed significantly higher (p < 0.05) antioxidant activity in GTE compared to BTE. However, TBARS values of uncured pork sausages significantly reduced (p < 0.05) for all levels of concentrations of BTE (0.05%, 0.10%, 020%, and 0.30%) and GTE (0.05%, 0.10, 0.20%, and 0.30%) during the 5 days storage period. The reduction of TBARS values for the BTE 0.05% treated sausage sample was not, however, significantly different (p > 0.05) to the BHT 0.10% treated sausage sample on fifth day of the storage period. The sensory evaluation of pork sausages incorporated with a BTE of 0.05% and 0.30%, GTE of 0.05% and the control were not significantly differences (p < 0.05) in color, odor, texture, juiciness, taste, or overall acceptability. Practical applications In the present study, we evaluated the antioxidant effect of green and black tea (Camellia sinensis L.) extracts in uncured pork sausages. Adding of black tea extract 0.05%, 0.30%, and green tea extract 0.05% can reduce the TBARS value in uncured pork sausages without altering color, odor, texture, juiciness, or overall acceptability. Therefore, 0.05%, 0.30% black tea extract and 0.05% green tea extracts can be considered as potential antioxidants in uncured pork sausages.

The chemical composition and sensory evaluation of yoghurt supplemented with green tea (GTE) or green coffee (GCE) extracts and its effect on reducing obesity complications in rats were investigated. Ethanol extracts were prepared from green tea and green coffee and their total phenolic compounds and identified phenolic compounds were evaluated. Results showed that the GTE and GCE have containing highly antioxidant activities. Yoghurt was prepared from low fat buffalo's milk (1.5% fat) blended with 1 % of GTE or GCE. Addition of GTE or GCE had significant effects on the chemical composition and sensory evaluation. Higher sensory scores were reported flavor for GTE and GCE yoghurt than control. It was also noticed that addition of GTE or GCE to yoghurt increased viscosity a key defensive effect on the gel-factors. Feeding of obese rats on yoghurt adding to GTE or GCE showed that significant differences reduced of weight compared to positive group. There are not found significant differences of heart weights tissue while, kidney and liver weights tissues were significant differences. Also, fed obese rats on yoghurt supplemented with GTE or GCE decreased the lipids profile comparative with positive control group and improvement of liver and kidney functions. The best treatment was observed in yoghurt adding to green coffee beans extract which had reduced value of liver and kidney function parameters followed by yoghurt adding to green tea extract. The histopathological examination observed normalized in the groups fed yoghurt supplemented green tea and coffee beans extracts. Supplemented yoghurt with green tea and coffee extracts could be increased the useful health effects of yogurt by intensifying antioxidant activity which had therapeutic effects on obese rats.

Chapter 46 - Green Tea Extract

Green and black teas extracts are known for their antibacterial activity against many pathogenic microorganisms. These studies have been necessitated by the need to combat the recent rise of drug-resistant human pathogens which is becoming a common occurrence in the world making easily manageable infections to become life threatening illnesses. This study evaluated the antimicrobial activity of water soluble green and black tea extracts from a high quality Kenyan tea clone TRFK 6/8 against antibiotic resistant Escherichia coli ATCC 25922 and Staphylococcus aureusATCC 25923 using agar well diffusion method. Green and black tea extracts effectively inhibited the growth of both E. coli ATCC 25922 and S. aureus ATCC 25923 at concentrations of 0.1 and 0.05 mg/ml, respectively after 24 h. Green tea extracts and gentamicin showed greater zone of inhibition compared to penicillin G. In addition, the possible synergistic activity of water soluble green tea extract and antibiotics was also determined using agar well diffusion method. A combination of penicillin G and green tea extract inhibited the growth of E. coli ATCC 25922 and S. aureus ATCC 25923 compared to penicillin G alone while gentamicin exhibited an additive and antagonistic effect depending on tested bacteria. Green and black tea extracts can be used as an antimicrobial agent and also green tea extract can be used in combination with penicillin G to manage resistant pathogenic bacteria. Furthermore, tea which is a proven safe, cheap and readily available compound can be used in more ambitious trials to test the antimicrobial efficacy and chemo-preventive effects in animal and human models. &nbsp; Key words: Green tea, black tea, antimicrobial activity, synergistic activity, gentamicin, penicillin G.

Metabolic syndrome (METS) consists of several independent risk factors for cardiovascular disease (CVD), one of which is vascular calcification (VC). Increased oxidative stress is essential in the pathogenesis of CVD in METS. One of the pathways involved in the pathogenesis of VC is the AKT pathway. Green tea and green coffee have many health benefits, including treating METS risk factors. Although several benefits of green tea and green coffee are known, there is not much information regarding the effects of these 2 extracts for treating heart disease, which is often found in METS sufferers. This research explores the benefits of green tea and green coffee extracts in preventing VC in METS through the AKT/MTOR mechanism. This research focuses on the mechanism of CVD in the AKT-mTOR, RUNX activity, and osteopontin (OPN) expression as one of the downstream pathogenesis of CVD in METS. The research was conducted on METS model rat treated with metformin and green tea with green coffee extracts. Male Sprague-Dawley rats were given a diet high in fat and sugar until they met METS for 4 months, then given treatment for 9 weeks. After 9 weeks, the rat had their aortas isolated for staining with specific antibodies to AKT1, MTOR, RUNX2, and serum OPN levels were measured using ELISA-sandwich methods. There was an increase in the expression of AKT1 and MTOR and a decrease in RUNX2 and OPN2, which was better than rat not treated with green tea and green coffee (p &lt; 0.05). A significant reduction was found in the green tea and green coffee therapy group, which was better than when given metformin alone (p &lt; 0.05). The results of this research showed that there is good potential for therapy using green tea and green coffee extracts to prevent VC in metabolic syndrome models. HIGHLIGHTS Metabolic syndrome consists of several independent risk factors for cardiovascular disease, one of which is vascular calcification. Increased oxidative stress is essential in the pathogenesis of cardiovascular disease in metabolic syndrome. This study explores the benefits of green tea and green coffee extracts in preventing vascular calcification in the metabolic syndrome model through the AKT/MTOR mechanism. This research focuses on the mechanism of CVD in the AKT-mTOR, RUNX activity, and osteopontin (OPN) expression as one of the downstream patogenesis of CVD in METS. The research was conducted on METS model rats treated with metformin and green tea with green coffee extracts. In conclusion, this study showed the potential for therapeutic use of green tea and green coffee extracts to prevent vascular calcification in rat models of metabolic syndrome by increasing AKT1 expression, decreasing MTOR and RUNX2, with the main implication of reducing the circular protein product, osteopontin. GRAPHICAL ABSTRACT

Selenium (Se) is one of the essential trace elements for human, but the Se deficiency of most areas resulting in low a Se content in fruits. To increase the Se uptake in fruit trees, the effects of foliar spraying four types of tea (black, green, white, and dark) infusions on the growth and Se uptake of Cyphomandra betacea were studied using a pot experiment. The white and dark tea infusions increased the biomass, superoxide dismutase activity, and catalase activity of C. betacea, while the black and green tea infusions had no significant effects. The green, white, and dark tea infusions increased the contents of photosynthetic pigments in C. betacea, the while the black tea infusions had no significant effects. The black, white, and dark tea infusions increased the peroxidase activity of C. betacea, while the green tea infusion decreased. All tea infusions increased the potassium (K) content in aboveground part of C. betacea and phosphorus (P) content in leaves to some extent. The different tea infusions decreased the Se contents in roots and shoots of C. betacea. Compared to the control, the black, green, white, and dark tea infusions decreased the Se contents in shoots by 16.67%, 27.30%, 33.33%, and 40.07%, respectively. Correlation analysis showed that the contents of P and K were negatively correlated with the Se content. Therefore, foliar spraying tea infusion can decrease the Se uptake in C. betacea, and the white and dark tea infusions can promote C. betacea growth.

This study aimed to evaluate the effects of the separate provision of green and oregano tea extracts on the biomarkers of the redox state and health condition in pre-weaned Jersey calves from birth to 60 days of life. Two experiments following the complete randomized design with measures repeated in time were carried out using 38 Jersey calves (17 and 21 calves in experiments 1 and 2, respectively). Calves were distributed according to date of birth into one of three groups: control (CON) - with no addition of extracts; oregano extract (OE) - addition of 70 mg of oregano extract/kg of body weight (BW) and green tea extract (GT) - addition of 35 mg of green tea extract/kg of BW. Eight biomarkers of the redox state were evaluated on days 1, 30, and 60 after birth, and variables measured on day 1 were used as covariates. Body temperature and occurrence of diarrhea were evaluated every two days. Regarding the main results, the supply of oregano extract reduced the concentration of oxidizing biomarkers, such as DCFP (oxidation of dichlorofluorescein in plasma) and carbonyl, and increased the activity of antioxidant enzymes, such as GPx and catalase. Green tea extract only reduced DCFP and tended to improve catalase activity. Calves remained healthy (no fever and only a few days with diarrhea), and plant extracts did not improve their health condition. The addition of green tea and oregano extracts into the diet has a positive effect on redox status in pre-weaned Jersey calves.

Obesity is the excess fat content in the body caused by the expansion of white adipose tissue. This condition begins with the differentiation of adipose tissue that is known to be controlled by 2 main transcription factors, such as Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) and CCAAT Enhancer-Binding Protein-α (C/EBP-α). Their activation and collaboration are critical for developing functioning adipocytes and preserving metabolic balance in adipose tissue. Research on the benefits of natural bioactive components that regulate adipogenesis has recently become an exciting focus. Foodstuffs reported to affect this condition positively include green coffee and green tea. The primary substance in green coffee is chlorogenic acid (CGA), meanwhile in green tea is epigallocatechin gallate (EGCG). Based on medical benefit potential, the present study evaluated the dose that produced the best effect on PPAR-γ and C/EBP-α expression between single and combined doses compared to undifferentiated adipocytes (3T3-L1). 3T3-L1 were cultured and divided into negative (NEG) and positive (DIF) groups. The DIF group is obtained by induction of differentiation, then divided into seventeen therapeutic doses. At the end of therapy, cells were fixed to measure the expression of PPAR-γ and C/EBP-α using the immunocytochemistry method. The DIF group produced the highest expression of PPAR-γ and C/EBP-α. Among the single-dose group, the lowest PPAR-γ was found in C320 and C/EBP-α in the T320 group. Meanwhile, the dose with the lowest PPAR-γ and C/EBP-α expression was found in the combination of green tea and coffee (TC) 160/80 (p-value = 0.00). The findings of this study showed that a combination of green tea and coffee extracts at doses of 160/80 has the potential for anti-obesity by suppressing the differentiation of 3T3-L1 into adipocytes by reducing the expression of PPAR-γ and C/EBP-α. HIGHLIGHTS Obesity is the excess fat content in the body caused by the expansion of white adipose tissue. This condition begins with the differentiation of adipose tissue that is known to be controlled by Peroxisome Proliferator-Activated Receptor-γ (PPAR-γ) and CCAAT Enhancer-Binding Protein-α (C/EBP-α) Natural bioactive components that regulate adipogenesis in green coffee are chlorogenic acid (CGA) and epigallocatechin gallate found in green tea (EGCG). Based on this potential, the present study evaluated the dose that produced the best effect on PPARɣ and C/EBP-α protein expression between single and combined doses compared to undifferentiated adipocytes (3T3-L1) Based on this study, the differentiated group of adipocytes (DIF) produced the highest expression of PPAR-γ and C/EBP-α. Among the single dose group, which showed the lowest average value of PPARɣ was C320, and for C/EBP-α was the T320 group. Meanwhile, the dose with the lowest PPAR-γ and C/EBP-α expression came from the combination of green tea and coffee (TC) 160/80 (p-value = 0.00) So, this research shows that green tea and green coffee extract significantly affected the expression of PPAR-γ and C/EBP-α. Green tea and coffee extracts at doses of TC 160/80 have the potential for anti-obesity by suppressing the differentiation of 3T3-L1 into adipocytes by reducing the expression of PPAR-γ and C/EBP-α GRAPHICAL ABSTRACT

Context: Dysregulation of glucose metabolism in metabolic syndrome (METS) is allegedly due to the disruption of insulin as the main pathway in cellular metabolism. Green tea and green coffee are known to have potential benefit in METS therapy. Aims: To evaluate the effect of therapy using decaffeinated green tea-green coffee extract as metformin's add-on in the risk factors of METS and its effect on the cardiac insulin-gene-related pathway, such as IRS1, PI3Kr1, and GLUT4. Methods: METS model rats were divided into five groups. The rats' level of body weight (BW), fasting blood glucose (FBG), triglycerides (TG), high-density lipoprotein (HDL), insulin (INS), and homeostatic model assessment for insulin resistance (HOMA-IR) were measured periodically. After nine weeks of treatment, the rats were euthanized, and the heart was isolated for measurement of IRS1, PI3Kr1, and GLUT4 gene expression by reverse-transcriptase polymerase chain reaction. Results: This study found that there was a decrease in BW, FBG, TG and an increase in HDL in METS model rats given therapy with metformin and green tea-green coffee extract (COMB) (p&lt;0.0001). There is also improvement in insulin resistance by reducing HOMA-IR in the COMB group (p = 0.0434 for INS and p&lt;0.0001 for HOMA-IR). This study found that IRS1, PI3K, and GLUT4 gene expression increased in the COMB group. The five groups differ significantly, with a p = 0.000. Conclusions: Therapy using a combination of decaffeinated green tea and green coffee extract as an add-on of metformin improved METS risk factor via a significant reduction in BW, FBG, TG, increasing HDL and improving insulin resistance. It also increased the IRS1, PI3Kr1, and GLUT4 gene expression as markers of cardiac insulin-gene-related pathways.