ATP citrate lyase

HIF Activation Suppressed by Garcinia Extract and HCA/Hydroxycitric acid Administration in Vitro [2]
The murine retinal cone cell line (661W) and the human RPE cell line (ARPE19) were used to evaluate HIF activity with a luciferase assay since photoreceptors and RPE cells significantly contribute the pathogenesis of AMD even though organoids or differentiated cells derived from iPS cells of AMD patients may be considered for better in vitro systems. Under a hypoxic condition, the activity of HIFα prolyl hydroxylase (PHD) decreases, which results in HIFα stabilization [12]. CoCl2 was added to stabilize the inhibition of PHD and to activate HIF signaling. Chetomin was used as a positive control of the HIF inhibitor. We used Garcinia extract. Table 1 lists its components showing that HCA accounts for more than half of the extract. Garcinia extract and HCA showed an HIF inhibitory effect compared with the control group in ARPE19 cells (Figure 1A) and 661W cells (Figure 1B).[2]

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Administration of Garcinia Extract and HCA/Hydroxycitric acid Downregulated Hif1a and Downstream Genes [2]
We examined how Garcinia extract and HCA affect mRNA expression of Hif1a and the downstream genes. In ARPE19 cells, Hif1a was significantly downregulated by administration of Garcinia extract regardless of the presence or absence of CoCl2 (Figure 2A). The downstream genes of HIFs such as Vegfa, Bnip3, and Pdk1, were upregulated by CoCl2 and significantly downregulated by Garcinia extract administration (Figure 2B–D). Similarly, Hif1a was downregulated by administration of Garcinia extract in 661W cells (Figure 2E). CoCl2-induced upregulation of Vegfa was also downregulated by Garcinia extract administration in 661W cells (Figure 2F). Expression of other downstream genes of HIFs showed a tendency to be downregulated as well as Vegfa (Figure 2G,H). HCA also downregulated Hif1a and the downstream genes in ARPE19 cells (Figure 3A–D) and 661W cells (Figure 3E–H). Both Garcinia extract and HCA suppressed HIF-1α protein expression increased by CoCl2 administration in ARPE19 cells (Figure 4A,B) and 661W cells (Figure 4C,D).

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Hydroxycitric acid (HCA) is one of the proven natural antiobesity agents enriched in the fruits of Garcinia gummi-gutta (L.) Roxb. (Family: Clusiaceae). The present research work was carried out to evaluate the genetic variability among 35 candidate plus trees (CPTs) using HCA estimated through HPLC and start codon targeted (SCoT) molecular markers. The association analysis between phenotypic and genotypic traits was also conducted. The selected CPTs showed an average HCA content of 29.11 mg/g and Gar 17 had the highest (48.32 mg/g) followed by Gar 6 (45.48 mg/g). SCoT marker analysis revealed that 19 primers, out of 30 yielded a total of 151 bands with 66.89% polymorphic bands. Principal coordinate analysis (PCoA) organized the CPTs into the four quadrants of a scatterplot irrespective of HCA content. Dendrogram based on neighbour joining method proved its reproducibility by its bootstrapping values, and it has three clusters. STRUCTURE analysis opened the probability of two assumed subpopulations within the selected individuals. Association analysis based on a general linear model (GLM) agreed with the strong association of SCoT 5d allele with HCA content, which also supports the promising nature of Gar 6 as per previous findings. Analysis based on HCA and SCoT markers was effective in tracing out the genetic variabilities among the CPTs and the marker-trait association. The findings are the first in G. gummi-gutta, best of our knowledge [5].

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Background/aims: (-)-Hydroxycitric acid (HCA) had been shown to suppress fat accumulation in animals and humans, while the underlying biochemical mechanism is not fully understood, especially little information is available on whether (-)-HCA regulates energy metabolism and consequently affects fat deposition. Methods: Hepatocytes were cultured for 24 h and then exposed to (-)-HCA (0, 1, 10, 50 µM), enzyme protein content was determined by ELISA; lipid metabolism gene mRNA levels were detected by RT-PCR. Results: (-)-HCA significantly decreased the number and total area of lipid droplets. ATP-citrate lyase, fatty acid synthase and sterol regulatory element binding protein-1c mRNA level were significantly decreased after (-)-HCA treatment, whereas peroxisome proliferator-activated receptor α mRNA level was significantly increased. (-)-HCA significantly decreased ATP-citrate lyase activity and acetyl-CoA content in cytosol, but significantly increased glucose consumption and mitochondrial oxygen consumption rate. (-)-HCA promoted the activity/content of glucokinase, phosphofructokinase-1, pyruvate kinase, pyruvate dehydrogenase, citrate synthase, aconitase, succinate dehydrogenase, malate dehydrogenase, NADH dehydrogenase and ATP synthase remarkably. Conclusions: (-)-HCA decreased lipid droplets accumulation by reducing acetyl-CoA supply, which mainly achieved via inhibition of ATP-citrate lyase, and accelerating energy metabolism in chicken hepatocytes. These results proposed a biochemical mechanism of fat reduction by (-)-HCA in broiler chickens in term of energy metabolism.[9]

The current research aimed to explore the impact of (-)-hydroxycitric acid (HCA) on fat metabolism and investigate whether this action of (-)-HCA was associated with modulation of glucose-6-phosphote isomerase (GPI) expression in chicken embryos. We constructed a recombinant plasmid (sh2-GPI) to inhibit GPI expression, and then embryos were treated with (-)-HCA. Results showed that (-)-HCA reduced lipid droplet accumulation, triglyceride content, and lipogenesis factors mRNA level and increased lipolysis factors mRNA expression, while this effect caused by (-)-HCA was markedly reversed when the chicken embryos were pretreated with sh2-GPI. (-)-HCA increased phospho (p)-acetyl-CoA carboxylase, enoyl-CoA hydratase short chain-1, carnitine palmitoyl transferase 1A, p-AMP-activated protein kinase, and peroxisome proliferators-activated receptor α protein expression, and this action of (-)-HCA also dispelled when the chicken embryos were pretreated with sh2-GPI. These data demonstrated that (-)-HCA decreased fat deposition via activation of the AMPK pathway, and the fat-reduction action of (-)-HCA was due to the increasing of GPI expression in chicken embryos.[8]

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HCA/Hydroxycitric acid treatment could reduce markers of renal impairment (Blood Urea Nitrogen and serum creatinine). There was significantly less calcium oxalate crystal deposition in mice treated with HCA. Calcium oxalate crystals induced the production of reactive oxygen species and reduced the activity of antioxidant defense enzymes. HCA attenuated oxidative stress induced by calcium oxalate crystallization. HCA had inhibitory effects on calcium oxalate-induced inflammatory cytokines, such as MCP-1, IL- 1 β, and IL-6. In addition, HCA alleviated tubular injury and apoptosis caused by calcium oxalate crystals.[1]

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Administration of Hydroxycitric acid/HCA Suppressed CNV Volume in the Model Mice [2]
HCA suspended in corn oil was injected intraperitoneally at 30 mg/kg/day for a total of two weeks, and the mice were irradiated with a laser one week after beginning the injections. The volume of CNV on the seventh day of the irradiation was significantly reduced in the HCA administration group when compared with the control group (Figure 6A,B).

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Administration of Hydroxycitric acid/HCA Suppressed HIF-1α Expression in Vivo [2]
HCA suspended in corn oil was intraperitoneally administered (30 mg/kg/day) to the mice for a total of 10 days, and the mice were irradiated with a laser on the seventh day of administration. In the retina and the choroid of the mice on the third day of irradiation, HIF-1α increased due to the laser irradiation and suppressed due to the administration of HCA (Figure 7A,B) even though the signal with the RPE/choroid tissue was weak.

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A total of 135 subjects were randomized to either active Hydroxycitric acid (n = 66) or placebo (n = 69); 42 (64%) in the active hydroxycitric acid group and 42 (61%) in the placebo group completed 12 weeks of treatment (P = .74). Patients in both groups lost a significant amount of weight during the 12-week treatment period (P<.001); however, between-group weight loss differences were not statistically significant (mean [SD], 3.2 [3.3] kg vs 4.1 [3.9] kg; P = .14). There were no significant differences in estimated percentage of body fat mass loss between treatment groups, and the fraction of subject weight loss as fat was not influenced by treatment group [4].

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The current research aimed to explore the impact of (-)-Hydroxycitric acid (HCA) on fat metabolism and investigate whether this action of (-)-HCA was associated with modulation of glucose-6-phosphote isomerase (GPI) expression in chicken embryos. We constructed a recombinant plasmid (sh2-GPI) to inhibit GPI expression, and then embryos were treated with (-)-HCA. Results showed that (-)-HCA reduced lipid droplet accumulation, triglyceride content, and lipogenesis factors mRNA level and increased lipolysis factors mRNA expression, while this effect caused by (-)-HCA was markedly reversed when the chicken embryos were pretreated with sh2-GPI. (-)-HCA increased phospho (p)-acetyl-CoA carboxylase, enoyl-CoA hydratase short chain-1, carnitine palmitoyl transferase 1A, p-AMP-activated protein kinase, and peroxisome proliferators-activated receptor α protein expression, and this action of (-)-HCA also dispelled when the chicken embryos were pretreated with sh2-GPI. These data demonstrated that (-)-HCA decreased fat deposition via activation of the AMPK pathway, and the fat-reduction action of (-)-HCA was due to the increasing of GPI expression in chicken embryos.[8]

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(-)-Hydroxycitric acid (HCA), a major active ingredient of Garcinia Cambogia extracts, had shown to suppress body weight gain and fat accumulation in animals and humans. While, the underlying mechanism of (-)-HCA has not fully understood. Thus, this study was aimed to investigate the effects of long-term supplement with (-)-HCA on body weight gain and variances of amino acid content in rats. Results showed that (-)-HCA treatment reduced body weight gain and increased feed conversion ratio in rats. The content of hepatic glycogen, muscle glycogen, and serum T4 , T3 , insulin, and Leptin were increased in (-)-HCA treatment groups. Protein content in liver and muscle were significantly increased in (-)-HCA treatment groups. Amino acid profile analysis indicated that most of amino acid contents in serum and liver, especially aromatic amino acid and branched amino acid, were higher in (-)-HCA treatment groups. However, most of the amino acid contents in muscle, especially aromatic amino acid and branched amino acid, were reduced in (-)-HCA treatment groups. These results indicated that (-)-HCA treatment could reduce body weight gain through promoting energy expenditure via regulation of thyroid hormone levels. In addition, (-)-HCA treatment could promote protein synthesis by altering the metabolic directions of amino acids [11].

Luciferase Assay [2]
We performed a luciferase assay using 661W and ARPE19, which were both transfected HIF-luciferase reporter gene constructs. These constructs encode the firefly luciferase gene under the control of HRE, which bind HIFs as previously described. As an internal control, these cells were co-transfected with a CMV-renilla luciferase construct. We seeded cells in 0.8 × 104 cells/well/70 μL in HTS Transwell®-96 Receiver Plate, White, TC-Treated, Sterile. At 24 h after seeding, HIF-αs were induced by 200 μM CoCl2. Garcinia extract (Garcinia Cambogia Extract 50% (Table 1)) and Hydroxycitric acid/HCA were dissolved in dimethyl sulfoxide and added into the growth medium at the same time as CoCl2. We added each compound dissolved in DMSO to the cell medium so that its concentration was 1 mg/mL considering the toxicity of the material. After the administration, cells were incubated for 24 h at 37 °C in a 5% CO2 incubator. Quantitation of the luciferase expression was performed using the Dual-Luciferase® Reporter Assay System. The fluorescent intensity was read by a microplate reader. Additionally, 100 nM of chetomin was used as a positive control for an HIF inhibitor and a DMSO-containing medium was used without CoCl2, Garcinia extract, and HCA as a vehicle control.

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Western Blot [2]
For in vitro experiments, we added 200 μM CoCl2, 1 mg/mL Garcinia extract, and Hydroxycitric acid/HCA to ARPE19 cell line while considering the toxicity of the material. Six hours after the administration, cells were collected in the RIPA buffer and mixed with protease inhibitors and MG132. Then, the cells were homogenized. Afterward, we centrifuged the samples (14,800 rpm, 4 °C, 30 min) and collected the supernatant. The protein concentration was adjusted to 75 μg/30 μL.

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Estimation of Hydroxycitric acid/HCA [5]
HCA from the dried fruit rinds of 35 CPTs of G. gummi-gutta (Babu et al., 2021; Vishnu et al., 2022) were extracted and purified according to Jayaprakasha and Sakariah (2000). The amount of HCA in 35 CPTs of G. gummi-gutta was estimated by HPLC. The Chromatography system consists of a UFLC (Shimadzu Corporation, Kyoto, Japan), SPD- 20A detector, communication bus module (CBM 20A), and Shim-pack GIST C18 column (5 μm, 4.6 ID × 250 mm). Detection of HCA was done at 0.1 AUFS sensitivity at 210 nm. The flow rate was set at 0.7 ml/ min under isocratic conditions using 0.6 mM sulfuric acid as mobile phase. HCA purified from potassium hydroxy citrate was used as the standard. Using 0.45 μm PTFE Syringe Filters (I-131 M Axiva; Sonipat, India), standard, and samples were filtered and injected into the system using a 20 μl injection syringe. A calibration curve was prepared by plotting varying HCA concentrations (200, 400, 600, 800, and 1000 mg/l) versus peak area. Concentrations of HCA in selected CPTs were estimated by the peak integration method and expressed as mg/g of sample.

Male C57BL/6J mice were divided into a control group, glyoxylate(GOX) 100 mg/kg group, a GOX+HCA 100 mg/kg group, and a GOX+HCA/Hydroxycitric acid 200 mg/kg group. Blood samples and kidney samples were collected on the eighth day of the experiment. We used Pizzolato staining and a polarized light microscope to examine crystal formation and evaluated oxidative stress via the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the expression of monocyte chemotactic protein-1(MCP-1), nuclear factor-kappa B (NF κB), interleukin-1 β (IL-1 β) and interleukin-6 (IL-6) messenger RNA (mRNA). The expression of osteopontin (OPN) and a cluster of differentiation-44(CD44) were detected by immunohistochemistry and qRT-PCR. In addition, periodic acid Schiff (PAS) staining and TUNEL assay were used to evaluate renal tubular injury and apoptosis. [1]

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Administration of Garcinia Extract and Hydroxycitric acid/HCA to Mice [2]
An MF diet mixed with Garcinia extract at a concentration of 0.2% was administered to 4-week-old male mice for a total of 7 weeks while considering the toxicity of the material. The control group was administered an MF diet. The mice were irradiated with a laser 6 weeks after beginning administration. We injected 30 mg/kg/day HCA suspended in corn oil intraperitoneally to 6-week-old male mice 5 days/week for a total of 2 weeks. The control group was injected with corn oil. The laser was irradiated 1 week after the initial injection.

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Medications in vivo [3]
Hydroxycitric acid tripotassium/K-HCA and citrate acid tripotassium/K-CA were tested as inhibitors to prevent the formation of stones. We randomly divided 600 flies into 6 groups (100 flies in each group) and fed them high-oxalate (0.05% NaOx) medium. Different concentrations (0.01%, 0.1%, and 1%) of K-HCA and K-CA were added in the medium of corresponding groups. Stone formation and life span were assessed in the same way as above.

In safety studies, we conducted acute oral toxicity, acute dermal toxicity, primary skin irritation, and primary eye irritation tests on animals using different doses of HCA-SX. The results showed that the LD50 of HCA-SX was greater than 5000 mg/kg after a single oral administration to fasted male and female albino rats. No significant toxicological symptoms were observed under the experimental conditions. In summary, these in vivo toxicological studies indicate that HCA-SX is a safe and natural dietary supplement under the tested conditions. Furthermore, HCA-SX can inhibit the uptake of [3H]-5-HT in isolated rat cerebral cortex slices (and increase 5-HT availability) in a manner similar to SRRIs, thus potentially aiding in appetite control and the treatment of depression, insomnia, migraines, and other serotonin deficiencies. [10]

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Adverse events[1]
No patients withdrew from the study due to treatment-related adverse events, and there was no significant difference in the number of adverse events reported between the placebo group and the treatment group (e.g., headache, 12 and 9, respectively; upper respiratory symptoms, 13 and 16, respectively; gastrointestinal symptoms, 6 and 13, respectively).

[1]. Hydroxycitric acid inhibits renal calcium oxalate deposition by reducing oxidative stress and inflammation. Curr Mol Med. 2020;20(7):527-535.

[2]. Therapeutic Effect of Garcinia cambogia Extract and Hydroxycitric Acid Inhibiting Hypoxia-Inducible Factor in a Murine Model of Age-Related Macular Degeneration. Int J Mol Sci. 2019 Oct 11;20(20). pii: E5049.

[3]. Hydroxycitric Acid Tripotassium Inhibits Calcium Oxalate Crystal Formation in the Drosophila Melanogaster Model of Hyperoxaluria. Med Sci Monit. 2019 May 17;25:3662-3667.

[4]. Garcinia cambogia (hydroxycitric acid) as a potential antiobesity agent: a randomized controlled trial. JAMA. 1998 Nov 11;280(18):1596-600.

[5]. Start codon targeted (SCoT) variability analysis and its association with hydroxy citric acid (HCA) in Garcinia gummi-gutta (L.) Roxb. Plant Gene, Volume 34, June 2023, 100415.

[6]. Chemistry and Biochemistry of (−)-Hydroxycitric Acid from Garcinia. J. Agric. Food Chem. 2002, 50, 1, 10-22.

[7]. Effects of garcinia cambogia (Hydroxycitric Acid) on visceral fat accumulation: a double-blind, randomized, placebo-controlled trial. Curr Ther Res Clin Exp. 2003 Sep;64(8):551-67.

[8]. (-)-Hydroxycitric Acid Influenced Fat Metabolism via Modulating of Glucose-6-phosphate Isomerase Expression in Chicken Embryos. J Agric Food Chem. 2019 Jul 3;67(26):7336-7347.

[9]. (-)-Hydroxycitric Acid Reduced Lipid Droplets Accumulation Via Decreasing Acetyl-Coa Supply and Accelerating Energy Metabolism in Cultured Primary Chicken Hepatocytes. Cell Physiol Biochem. 2017;43(2):812-831.

[10]. Safety and mechanism of appetite suppression by a novel hydroxycitric acid extract (HCA-SX). Mol Cell Biochem. 2002 Sep;238(1-2):89-103.

[11]. (-)-Hydroxycitric Acid Nourishes Protein Synthesis via Altering Metabolic Directions of Amino Acids in Male Rats. Phytother Res. 2016 Aug;30(8):1316-29.

Garcinia cambogia is a carbonyl compound.
See also: Garcinia gummi-gutta fruit (partial).
Hydroxycitric acid is a carbonyl compound.
Hydroxycitric acid has been reported in Garcinia cowa, Hibiscus sabdariffa, and Garcinia atroviridis, with relevant data.
See also: Hydroxycitric acid (note moved to).
Background: Age-related macular degeneration (AMD) is a leading cause of blindness and can be divided into two types: atrophic AMD (dry AMD) and neovascular AMD (wet AMD). Dry AMD is characterized by cellular degeneration of the retinal pigment epithelium, choroidal capillaries, and photoreceptor cells. Wet AMD is characterized by the invasion of abnormal choroidal vessels. Although anti-vascular endothelial growth factor (VEGF) therapy has significant therapeutic effects on this disease, long-term potent VEGF antagonism may lead to choroidal and retinal atrophy and systemic adverse reactions. We focused on the regulation of VEGF transcription by hypoxia-inducible factor (HIF) and reported the inhibitory effect of HIF inhibitors on ocular phenotypes in animal models. Many known HIF inhibitors are classified as anticancer drugs, but their systemic side effects are a concern in clinical applications. This study explored food components with HIF inhibitory effects and validated their efficacy in an AMD animal model. Methods: Food components were screened using luciferase reporter gene assay. C57BL6/J mice were administered Garcinia Cambogia extract (Garcinia Cambogia extract) and hydroxycitric acid (HCA), respectively. Laser irradiation induced choroidal neovascularization (CNV). Results: Both Garcinia Cambogia extract and HCA showed inhibitory effects on hypoxia-inducible factor (HIF) in luciferase assays. In laser-induced CNV model mice, CNV volume was significantly reduced after treatment with Garcinia Cambogia extract and HCA. Conclusion: Garcinia Cambogia extract and HCA showed therapeutic effects in a mouse model of age-related macular degeneration (AMD).
Keywords: Garcinia cambogia; age-related macular degeneration; choroid; hydroxycitric acid; hypoxia-inducible factor; laser-induced angiogenesis; retina. [2]

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Background: Hydroxycitric acid is a potential lithotripter for calcium oxalate (CaOx) kidney stones. This study aimed to evaluate the safety and efficacy of tripotassium hydroxycitric acid (K-HCA) against CaOx crystal formation using a Drosophila hyperoxaluria model. Materials and Methods: Wild-type Drosophila melanogaster were fed standard culture medium supplemented with ethylene glycol or sodium oxalate to induce hyperoxaluria. Malpighian tubules were dissected every 3 days and observed under a microscope. Crystal deposition scores of each Malpighian tubule were evaluated at 200x magnification. Using a Drosophila hyperoxaluria model, we investigated the inhibitory effects of tripotassium hydroxycitric acid and tripotassium citrate (K-CA) on calcium oxalate crystal formation. Survival rates of each group were also assessed. Results: Feeding with 0.05% sodium oxalate significantly increased the formation of calcium oxalate in Malpighian tubules, but did not shorten the lifespan of fruit flies. Therefore, we selected the 0.05% sodium oxalate-induced hyperoxaluria model for subsequent studies. After treatment, stone scores showed that both K-CA and K-HCA significantly inhibited calcium oxalate crystal formation in a dose-dependent manner, and at a lower dose (0.01%), K-HCA was more effective than K-CA. Furthermore, the lifespan of fruit flies in all groups remained unchanged after K-CA or K-HCA treatment, indicating its safety. Conclusion: The hyperoxaluria model of fruit flies fed a 0.05% sodium oxalate diet may be an effective tool for screening new drugs to treat calcium oxalate stones. K-HCA may be a promising drug for the prevention of calcium oxalate stones, with satisfactory efficacy and acceptable safety. [3] Background: Hydroxycitric acid is the active ingredient in Garcinia Cambogia, which competitively inhibits the mitochondrial extramitochondrial enzyme pro-3S lyase. As a citrate lyase that may play an important role in inhibiting de novo fat synthesis, Garcinia Cambogia is thought to reduce body weight and body fat. Objective: To evaluate the efficacy of Garcinia Cambogia in reducing body weight and body fat in overweight individuals. Design: A 12-week randomized, double-blind, placebo-controlled trial. Location: Outpatient Weight Control Research Center. Participants: Overweight male and female subjects (mean body mass index [weight (kg) divided by height (m) squared] approximately 32 kg/m²). Intervention: Subjects were randomly assigned to receive either the active herbal compound (1500 mg hydroxycitric acid daily) or a placebo, both groups were required to follow a high-fiber, low-energy diet. The treatment period was 12 weeks. Body weight was assessed every other week, and body fat was measured at week 0 and week 12.
Primary outcome measures: Changes in body weight and body fat mass.
Results: A total of 135 participants were randomly assigned to either the active hydroxycitric acid group (n = 66) or the placebo group (n = 69); 42 participants (64%) in both the active hydroxycitric acid group and the placebo group completed the 12-week treatment (P = 0.74). Both groups experienced significant weight loss during the 12-week treatment period (P < 0.001); however, there was no statistically significant difference in weight loss between the groups (mean [standard deviation], 3.2 [3.3] kg vs 4.1 [3.9] kg; P = 0.14). There was no significant difference in the estimated percentage reduction in body fat between the treatment groups, nor was the proportion of fat in the participants' weight loss affected by the treatment groups.
Conclusion: Garcinia Cambogia did not produce a more significant effect on weight loss and body fat reduction than the placebo group. [4]

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The moderate level of genetic diversity exhibited by the CPT of Garcinia gummi-guttata supports the precise amplification of SCoT markers and the presence of specific SCoT alleles within the species. The inter-individual relationships, regardless of hydroxycitric acid (HCA) content, are likely the result of gene migration. Gar1 is genetically distinct from other individuals, as can be seen from scatter plots and dendrograms. STRUCTURE analysis revealed the possible existence of two hypothetical subpopulations among the selected individuals. Gar17 had the highest HCA content. However, GLM analysis revealed promising results for germplasm Gar6, with the strongest correlation between its HCA content and the SCoT 5d allele being the best evidence. The corresponding allele can significantly affect the HCA content of this species and should be considered when implementing breeding strategies for this species. These findings can be extended to marker-assisted selection and genetic improvement of Garcinia gummi-guttata. [5]

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(−)-hydroxycitric acid [(−)-HCA] is the main acid in the pericarp of Garcinia cambogia, Garcinia indica, and Garcinia atroviridis. (−)-HCA has been shown to be a potent inhibitor of ATP citrate lyase (EC 4.1.3.8), which catalyzes the cleavage of extramitochondrial citrate into oxaloacetate and acetyl-CoA: citrate + ATP + CoA → acetyl-CoA + ADP + Pi + oxaloacetate. Inhibition of this reaction limits the availability of acetyl-CoA units required for fatty acid synthesis and lipogenesis during lipogenic diets (i.e., high-carbohydrate diets). Numerous animal studies have shown that (−)-HCA inhibits fatty acid synthesis, lipogenesis, food intake, and induces weight loss. In vitro studies have revealed its inhibitory effects on the synthesis of various precursor fatty acids and lipogenesis. However, some clinical study results are controversial. This review explores the literature on the following aspects: the origin of (−)-HCA; the discovery of (−)-HCA; the isolation, stereochemistry, properties, assay methods and derivatives of (−)-HCA; and its biochemical properties, including inhibition of citrate lyase, effects on fatty acid synthesis and lipogenesis, effects on ketone body production, other biological effects, possible mechanisms of action for reducing food intake, promotion of glycogenogenesis, gluconeogenesis and lipid oxidation, the role of (−)-HCA as a weight management agent, and some potential issues of (−)-HCA. It systematically elucidates the scattered literature on (−)-HCA and its possible mechanisms of action and inspires further research. [6]

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Background: (−)-hydroxycitric acid (HCA) is an active ingredient extracted from the pericarp of Garcinia cambogia. It inhibits citrate lyase and has been used to treat obesity. Objective: The primary endpoint of this study was to evaluate the effect of Garcinia cambogia extract on visceral fat accumulation after 12 weeks of administration. Secondary endpoints included body measurements (height, weight, body mass index [BMI], waist circumference, hip circumference, and waist-to-hip ratio) and laboratory parameters (total cholesterol, triglycerides, and free fatty acids). Methods: This study employed a double-blind, randomized, placebo-controlled, parallel-group design. Participants aged 20–65 years with a visceral fat area >90 cm² were enrolled. Participants were randomly assigned to receive either Garcinia Cambogia extract (containing 1000 mg HCA daily) or placebo for 12 weeks. At the end of treatment, both groups received placebo for 4 weeks to assess for rebound effects. Each participant underwent umbilical CT scans at -2, 0, 12, and 16 weeks. Results: Forty-four participants were randomized at baseline, and 39 completed the study (Garcinia Cambogia group, n=18; placebo group, n=21). At week 16, compared with the placebo group, the Garcinia Cambogia group showed significant reductions in visceral fat, subcutaneous fat, and total fat area (all P < 0.001). No serious adverse reactions were observed during the trial. At week 12, there were no significant differences in BMI and weight between the two groups, but male subjects showed a slight decrease in weight and BMI. No rebound effect was observed from week 12 to week 16. Conclusion: Garcinia Cambogia can reduce abdominal fat accumulation in patients with visceral fat accumulation obesity, regardless of gender. No rebound effect was observed. Therefore, it is expected that Garcinia Cambogia can help prevent and reduce the accumulation of visceral fat. [7]

C6H8O8

208.12292

529.885

27750-10-3

Hydroxycitric acid;6205-14-7

185620

White to off-white solid powder

1.947g/cm3

393.3ºC at 760mmHg

205.8ºC

8.17E-08mmHg at 25°C

1.619

-2.6

5

8

5

14

271

2

C(C(=O)O)[C@]([C@@H](C(=O)O)O)(C(=O)O)O

ZMJBYMUCKBYSCP-CVYQJGLWSA-N

InChI=1S/C6H8O8/c7-2(8)1-6(14,5(12)13)3(9)4(10)11/h3,9,14H,1H2,(H,7,8)(H,10,11)(H,12,13)/t3-,6+/m1/s1

(1S,2S)-1,2-dihydroxypropane-1,2,3-tricarboxylic acid

HYDROXYCITRATE, L-; 8W94T9026R; HYDROXYCITRIC ACID, (-)-; (1S,2S)-1,2-dihydroxy-1,2,3-propanetricarboxylic acid; Regulator; Haes cpd; Hydroxycitric acid ethylenediamine salt; ...; 27750-10-3;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~5 mg/mL (~24.02 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)