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Intestinal stem cells (ISCs) are essential to maintain intestinal epithelial regeneration and barrier function. Our previous work showed that glucomannan from Aloe vera gel (AGP) alleviated epithelial damage, but the mechanism was still elusive. Herein, RNA-sequencing analysis showed that proliferation and differentiation of intestinal epithelial cells as well as the canonical Wnt pathway were involved in this process. Further experiments exhibited that AGP promoted nuclear translocation of β-catenin and expression of transcription factor 7, increased the number of Lgr5+ ISCs, and differentiated epithelial cells in mice colon. Intriguingly, AGP reversed the inhibition of IEC-6 cells proliferation induced by an inhibitor of the canonical Wnt pathway. Hence, this study implied that AGP promoted proliferation and differentiation of colon stem cells via Wnt/β-catenin signaling, which subsequently facilitated the regeneration of epithelial cells and alleviated colitis in mice. It may provide new insights into the role of polysaccharides in regulating intestinal homeostasis and relieving intestinal injury.

Regeneration of epithelia is crucial for maintaining the intestinal barrier and homeostasis. Our previous work showed that exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) regulated the barrier function and homeostasis of the intestine; however, the relevant mechanisms remain obscure. Therefore, we sought to explore the role of EPS116 in promoting intestinal epithelial regeneration. Our data showed that the administration of EPS116 markedly ameliorated inflammatory bowel disease-related phenotypes and promoted the regeneration of crypts in the colon of colitis mice. The results of immunofluorescence and reverse transcription-quantitative polymerase chain reaction experiments indicated that EPS116 strikingly increased the number of intestinal stem cells (ISCs) and the expression of differentiation markers for goblet cells, enterocytes, and enteroendocrine cells in the mouse colon. Intestinal microbiota analysis showed that EPS116 increased microbial populations associated with intestinal regeneration and glycan metabolism. Therefore, the present study revealed a novel model that EPS116 promoted the intestinal homeostasis through modulating the proliferation and differentiation of ISCs and altering the gut microbiota profile.

Targeting autophagy and lysosome may serve as a promising strategy for cancer therapy. Tea polysaccharide (TP) has shown promising antitumor effects. However, its mechanism remains elusive. Here, TP was found to have a significant inhibitory effect on the proliferation of colon cancer line HCT116 cells. RNA-seq analysis showed that TP upregulated autophagy and lysosome signal pathways, which was further confirmed through experiments. Immunofluorescence experiments indicated that TP activated transcription factor EB (TFEB), a key nuclear transcription factor modulating autophagy and lysosome biogenesis. In addition, TP inhibited the activity of mTOR, while it increased the expression of Lamp1. Furthermore, TP ameliorated the lysosomal damage and autophagy flux barrier caused by Baf A1 (lysosome inhibitor). Hence, our data suggested that TP repressed the proliferation of HCT116 cells by targeting lysosome to induce cytotoxic autophagy, which might be achieved through mTOR-TFEB signaling. In summary, TP may be used as a potential drug to overcome colon cancer.

Water-soluble β-d-glucan was obtained from wild Cordyceps sinensis by alkali solution and ethanol precipitation. The structure characteristics were determined using high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD), methylation combined with gas chromatography-mass spectrometry, and one-/two-dimensional nuclear magnetic resonance spectroscopy. Results showed that β-d-glucan had a structure of every seven (1→3)-β-d-Glcp backbone residues with two (1→6)-β-d-Glcp branches. Additionally, conformation properties in different solvents were investigated by static light scattering, dynamic light scattering, and HPSEC with multiple detectors. It was found that β-d-glucan in 0.5 M NaOH had a narrow unimodal distribution of hydrodynamic radius displaying a spherical coil conformation, whereas it formed severe aggregation in dimethyl sulfoxide. In 0.1 M NaNO3, β-d-glucan mainly existed as a rod-like conformation corresponding to a helical structure together with small aggregates (10%). This work added more information to the understanding of C. sinensis polysaccharides.

Type 2 diabetes (T2D) is a pandemic disease chiefly characterized by hyperglycemia. In this study, the combination of serum lipidomic and metabolomic approach was employed to investigate the effect of arabinoxylan on type 2 diabetic rats and identify the critical biomarkers of T2D. Metabolomics analysis revealed that branched-chain amino acids, 12α-hydroxylated bile acids, ketone bodies, and several short- and long-chain acylcarnitines were significantly increased in T2D, whereas lysophosphatidylcholines (LPCs) were significantly decreased. Lipidomics analysis indicated T2D-related dyslipidemia was mainly associated with the increased levels of acetylcarnitine, free fatty acids (FFA), diacylglycerols, triacylglycerols, and cholesteryl esters and the decreased levels of some unsaturated phosphatidylcholines (less than 22 carbons). These variations indicated the disturbed amino acid and lipid metabolism in T2D, and the accumulation of incompletely oxidized lipid species might eventually contribute to impaired insulin action and glucose homeostasis. Arabinoxylan treatment decreased the concentrations of 12α-hydroxylated bile acids, carnitines, and FFAs and increased the levels of LPCs. The improved bile acid and lipid metabolism by arabinoxylan might be involved in the alleviation of hypercholesterolemia and hyperlipidemia in T2D.

Probiotic lactobacilli and their exopolysaccharides (EPS) are thought to modulate mucosal homeostasis; however, their mechanisms remain elusive. Thus, we tried to clarify the role of exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) in the intestinal mucosal homeostasis. Our results indicated that EPS116 regulated the colon mucosal healing and homeostasis, enhanced the goblet cell differentiation, and promoted the expression of Muc2 gene in vivo and in vitro. Further experiments showed that EPS116 promoted the expression and phosphorylation of transcription factor c-Jun and facilitated its binding to the promoter of Muc2. Moreover, knocking down c-Jun or inhibiting its function in LS 174T cells treated with EPS116 led to decreased expression of Muc2, implying that EPS116 promoted the colonic mucosal homeostasis and Muc2 expression via c-Jun. Therefore, our study uncovered a novel model where EPS116 enhanced colon mucosal homeostasis by controlling the epithelial cell differentiation and c-Jun/Muc2 signaling.

Diabetes and its complications are one of the most concerned metabolic diseases worldwide and threaten human health severely. Hypoglycemic and hypolipidemic effects of glucomannan extracted from konjac on high-fat diet and streptozocin-induced type 2 diabetic rats were evaluated in this study. Administration of konjac glucomannan significantly decreased the levels of fasting blood glucose, serum insulin, glucagon-like peptide 1, and glycated serum protein. The concentrations of serum lipids, including total cholesterol, triacylglycerols, low-density lipoprotein cholesterol, and non-esterified fatty acid, were notably reduced by konjac glucomannan treatment. In addition, antioxidant capacity, pancreatic injury, and adipose cell hypertrophy were ameliorated by konjac glucomannan administration in type 2 diabetic rats. Besides, ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based lipidomics analysis was used to explore the improvement of lipid metabolic by konjac glucomannan treatment. The disturbance of glycerolipid (diacylglycerol, monoacylglycerol, and triacylglycerol), fatty acyl (acylcarnitine and hydroxyl fatty acid), sphingolipid (ceramide and sphingomyelin), and glycerophospholipid (phosphatidylcholine) metabolism were attenuated by the glucomannan treatment. This study provided new insights for investigating the anti-diabetic effects of konjac glucomannan and suggests that konjac glucomannan may be a promising nutraceutical for treating type 2 diabetes.

The chain conformation of a β-glucan extracted from black fungus (BFP) was studied by static/dynamic light scattering, viscometry, atomic force microscopy (AFM), and molecular dynamics (MD) simulat...

Lactic acid fermentation represents a novel method to produce bioactive functional ingredients, including polysaccharides. In this work, a selected lactic acid bacteria strain NCU116 was used to ferment Asparagus officinalis (asparagus) pulps. Two polysaccharides were subsequently separated from both unprocessed and fermented asparagus pulps, namely, asparagus polysaccharide (AOP) and fermented-AOP (F-AOP). The physicochemical and bioactive properties of AOP and F-AOP were characterized and investigated. High-performance anion-exchange chromatography showed that fermentation increased the proportions of rhamnose, galacturonic acid, and glucuronic acid in polysaccharides by 46.70, 114.09, and 12.75‰, respectively. High-performance size-exclusion chromatography revealed that fermentation decreased the average molecular weight from 181.3 kDa (AOP) to 152.8 kDa (F-AOP). Moreover, the fermentation reduced the particle size and changed the rheology property. In vitro, F-AOP displayed superior free radical scavenging properties compared to AOP, using 2,2-diphenyl-1-picryhydrazyl, hydroxyl, and superoxide anion radical scavenging assays. In vivo, F-AOP administration dose-dependently promoted a gradual shift from Th17-dominant acute inflammatory response (IL-17 and RORγt) to Th1-dominant defensive immune response (IFN-γ and T-bet). These results indicated that the Lactobacillus plantarum NCU116 fermentation was practical and useful to obtain promising bioactive polysaccharides.

Lactic acid bacteria (LAB) and their exopolysaccharides (EPS) are recognized to promote intestinal barrier function by mechanisms that remain incompletely understood. Herein, we sought to identify the roles of exopolysaccharides from Lactobacillus plantarum NCU116 (EPS116) in intestinal barrier function. Our data showed that EPS116 attenuated dextran sodium sulfate (DSS) induced colitis and promoted epithelial barrier function and the expression of tight junction (TJ) proteins in vivo and in vitro. Moreover, chromatin immunoprecipitation data showed that EPS116 facilitated STAT3 (signal transducer and activator of transcription 3) binding to the promoter of occludin and ZO-1. Furthermore, knockdown of STAT3 in Caco-2 cell with EPS116 treatment led to decreased expression of occludin and ZO-1 and increased intestinal permeability, suggesting that the regulation of epithelial barrier function by EPS116 should be STAT3 dependent. Thus, our data revealed a novel mechanism that EPS116 inhibited intestinal inflammation via regulating intestinal epithelial barrier function.

Diabetes mellitus is a worldwide concern and obviously influences the quality of life of humans. Dietary polysaccharides were mainly from natural sources, namely, plants, fungi, algae, etc. They were resistant to human digestion and absorption, with complete or partial fermentation in the large bowel, and have shown antidiabetic ability. In this perspective, a literature search was conducted to provide information on the antidiabetic mechanism of dietary polysaccharides based on the whole gastrointestinal process, which was a new angle of view for understanding their antidiabetic mechanism. Further studies could take efforts on the mechanisms of the polysaccharide action through host-microbiota interactions targeting diabetes.

The interleukin-6 (IL-6)/signal transducer and activator of transcription (STAT)-3 signaling pathway regulates proliferation and survival of intestinal epithelial cells and has profound impact on the tumorigenesis of colitis-associated cancer (CAC). Tea polysaccharides (TPS) are the major nutraceutical component isolated from tea-leaves and are known to possess antioxidant, anti-inflammatory, and antitumor bioactivities. Here, we investigated the antitumor activities of TPS on CAC using the azoxymethane/dextran sulfate sodium (AOM/DSS) mouse model and IL-6-induced colorectal cancer cell line (CT26) and determined whether TPS exerted its antitumor effects through the IL-6/STAT3 pathway. Results demonstrated that TPS significantly decreased the tumor incidence, tumor size, and markedly inhibited the infiltration of pro-inflammatory cells and the secretion of pro-inflammatory cytokines via balancing cellular microenvironment. Furthermore, we found that TPS suppressed the activation of STAT3 and transcriptionally regulated the expressions of downstream genes including MMP2, cyclin Dl, survivin, and VEGF both in vivo and in vitro. Thus, it was concluded that TPS attenuated the progress of CAC via suppressing IL-6/STAT3 pathway and downstream genes' expressions, which indicated that TPS may be a hopeful antitumor agent for the prevention and treatment of colon cancer.

Our previous study has demonstrated that Ganoderma atrum polysaccharide (PSG-1) has immunomodulatory activity on spleen lymphocytes. However, how PSG-1 exerts its effect on purified lymphocytes is still obscure. Thus, this study aimed to investigate the immunomodulatory activity of PSG-1 on purified T lymphocytes and further elucidate the underlying mechanism based on RNA sequencing (RNA-seq). Our results showed that PSG-1 promoted T lymphocytes proliferation and increased the production of IL-2, IFN-γ, and IL-12. Meanwhile, RNA-seq analysis found 394 differentially expressed genes. KEGG pathway analysis identified 20 significant canonical pathways and seven biological functions. Furthermore, PSG-1 elevated intracellular Ca2+ concentration and calcineurin (CaN) activity and raised the p-ERK, p-JNK, and p-p38 expression levels. T lymphocytes proliferation and the production of IL-2, IFN-γ, and IL-12 were decreased by the inhibitors of calcium channel and mitogen-activated protein kinases (MAPKs). These res...

The present study was to evaluate the beneficial effect of polysaccharide isolated from Ganoderma atrum (PSG-1) on liver function in type 2 diabetic rats. Results showed that PSG-1 decreased the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), while increasing hepatic glycogen levels. PSG-1 also exerted strong antioxidant activities, together with upregulated mRNA expression of peroxisome proliferator-activated receptor-γ (PPAR-γ), glucose transporter-4 (GLUT4), phosphoinositide 3-kinase (PI3K), and phosphorylated-Akt (p-Akt) in the liver of diabetic rats. Moreover, the concentrations of short-chain fatty acids (SCFA) were significantly higher in the liver, serum, and faeces of diabetic rats after treating with PSG-1 for 4 weeks. These results suggest that the improvement of PSG-1 on liver function in type 2 diabetic rats may be due to its antioxidant effects, SCFA excretion in the colon from PSG-1, and regulation of hepatic glucose uptake by inducing GLUT4 translocation through PI3K/Akt signaling pathways.

This research was aimed to study the effect of Dendrobium officinale polysaccharide (Dendronan) on colonic health. Mice were fed Dendronan at doses of 40, 80, and 160 mg/kg body weight for 0, 10, 20, and 30 days, respectively. Results showed that Dendronan, which has a special structure formed by mannose and glucose, rich in O-acetyl groups, exhibited improving effects on colonic and fecal parameters of Balb/c mice. After Dendronan feeding, the content of short-chain fatty acids (SCFAs), colon length and index, and fecal moisture were increased, whereas colonic pH was decreased and defecation time was shortened. All of these changes were significantly different between polysaccharide-treated groups and the control group (p < 0.05). These findings suggested that an adequate intake of Dendronan is beneficial to the process of fermentation and regulation of colonic microenvironment, thus playing a role in the maintenance of colonic health.

The aim of the present study was to examine the role of Ganoderma atrum polysaccharide (PSG-1) in reactive oxygen species (ROS) generation and mitochondrial function in hyperglycemia-induced angiopathy. In this work, ROS scavenger, oxidizing agent tert-butylhydroperoxide (tBH), mitochondrial permeability transition pore (mPTP) blockers, and caspase inhibition are used to investigate whether PSG-1 may promote survival of human umbilical vein cells (HUVECs) through preventing the overproduction of ROS and mitochondrial dysfunction. Experimental results show that exposure of HUVECs to 35.5 mmol/L glucose increases the proportion of cells undergoing apoptosis. PSG-1, mPTP blocker, or caspase inhibition can reduce apoptosis and ROS generation. PSG-1 also increases mitochondrial Bcl-2 protein formation and mitochondrial membrane potential (ΔΨm) but inhibits Bax translocation, cytochrome c release, and caspase activation. In summary, vascular protection of PSG-1 can be mediated by a mitochondria-ROS pathway. ROS generation and mPTP induction are critical for high glucose-mediated apoptosis. PSG-1 ameliorates endothelial dysfunction by inhibiting oxidative stress and subsequent mitochondrial dysfunction.

The aim of this study was to investigate the molecular mechanism underlying the immunomodulatory effect of Ganoderma atrum polysaccharide (PSG-1) in spleen lymphocytes. Our results showed that PSG-1 increased the intracellular Ca2+ concentration and calcineurin (CaN) activity. Moreover, PSG-1 was found to elevate nuclear factor of activated T cells (NFAT) activity, but this effect could be diminished by the treatment of CaN inhibitors (cyclosporin A and FK506). PSG-1-induced interleukin (IL)-2 production was also inhibited by cyclosporin A and FK506. In addition, PSG-1 was found to significantly enhance protein kinase C (PKC) activity. PKC was involved in induction of NFAT activity by PSG-1, as evidenced by abrogation of NFAT activity by PKC inhibitor calphostin C, which significantly decreased PSG-1-induced IL-2 production. On the basis of these results, we concluded that PSG-1 may induce activation of spleen lymphocytes at least in part via the Ca2+/CaN/NFAT/IL-2 signaling pathway and the PKC/NFAT/IL-2 signaling pathway cooperatively regulated PSG-1-induced activation of spleen lymphocytes.

The effect of carrot juice fermented with Lactobacillus plantarum NCU116 on high-fat and low-dose streptozotocin (STZ)-induced type 2 diabetes in rats was studied. Rats were randomly divided into five groups: non-diabetes mellitus (NDM), untreated diabetes mellitus (DM), DM plus L. plantarum NCU116 (NCU), DM plus fermented carrot juice with L. plantarum NCU116 (FCJ), and DM plus non-fermented carrot juice (NFCJ). Treatments of NCU and FCJ for 5 weeks were found to favorably regulate blood glucose, hormones, and lipid metabolism in the diabetic rats, accompanied by an increase in short-chain fatty acid (SCFA) in the colon. In addition, NCU and FCJ had restored the antioxidant capacity and morphology of the pancreas and kidney and upregulated mRNA of low-density lipoprotein (LDL) receptor, cholesterol 7α-hydroxylase (CYP7A1), glucose transporter-4 (GLUT-4), peroxisome proliferator-activated receptor-α (PPAR-α), and peroxisome proliferator-activated receptor-γ (PPAR-γ). These results have for the first time demonstrated that L. plantarum NCU116 and the fermented carrot juice had the potential ability to ameliorate type 2 diabetes in rats.

Ganoderma atrum is one species of edible and pharmaceutical mushroom with various biological activities. Recently, a novel polysaccharide, PSG-1, was purified from G. atrum. The antitumor activity and its mechanism of action were studied. In vitro, PSG-1 has little effect on inhibiting proliferation of CT26 tumor cells. However, the tumor size was significantly decreased in PSG-1-treated mice. The results showed that PSG-1 induced apoptosis in CT26 cells. Moreover, the intracellular cyclic AMP (cAMP) level and protein kinase A (PKA) activity were markedly increased in PSG-1-treated mice. In contrast, the contents of cyclic GMP and DAG and the PKC activity were decreased. Similarly, the expression of PKA protein was upregulated, while PKC protein expression in PSG-1-treated group was lowered. Additionally, PSG-1 increased the immune organ index and serum biochemistry parameter. In general, PSG-1 enhances the antitumor immune response, induces apoptosis in CT26-bearing mice, and could be a safe and effective adjuvant for tumor therapy or functional food.

White rice porridge and mixed grain porridge, which are often consumed in many countries, were used as two models to evaluate the effects of gum arabic on glucose levels and microbial short-chain fatty acids (SCFA). Gum arabic was incorporated into the two porridges individually. Apparent viscosity of the two porridges was significantly increased, and their glucose productions during gastrointestinal digestion were notably lowered (p < 0.05). Diffused glucose amount was significantly decreased after gum arabic addition (p < 0.05). Furthermore, blood glucose rise after oral administration of porridges in mice was considerably lowered after fortified with gum arabic (p < 0.05). Microbial SCFA production during in vitro fermentation of porridges was significantly increased after gum arabic addition, which may also have beneficial effects on reducing postprandial glycemic response. Therefore, gum arabic may be a helpful ingredient, which could be added in porridges to have benefits for the reduction of postprandial glycemic response.

Polysaccharide from the seeds of Plantago asiatica L. was given via oral administration to mice (0.4 g/kg body weight, 30 days) to observe its effects on mouse nutrient metabolism and colon microbiota. It was found the polysaccharide intake could lower the apparent absorption of lipid. Total triglyceride, cholesterol, and atherogenic index in blood serum with total lipid and cholesterol levels in liver of polysaccharide group mice were all significantly lower than those of the control group (p < 0.05). Furthermore, the effect of the polysaccharide intake on mouse colon bacterial communities was investigated. Mice from the polysaccharide group showed a higher colon bacterial diversity than the control group. Bacteroides sp., Eubacterium sp., butyrate-producing bacteria Butyrivibrio sp., and probiotics Bifidobacterium bifidum , Lactobacillus fermentum , and Lactobacillus reuteri in mouse colon were all increased after polysaccharide intake. These indicated that the intake of polysaccharide from P. asiatica L. could be beneficial for lipid metabolism and colon microbiota.

Effects of microwave irradiation on microbial short-chain fatty acid production and the activites of extracellular enzymes during in vitro fermentation of the polysaccharide from Plantago asiatica L. were investigated in this study. It was found that the apparent viscosity, average molecular weight, and particle size of the polysaccharide decreased after microwave irradiation. Reducing sugar amount increased with molecular weight decrease, suggesting the degradation may derive from glycosidic bond rupture. The polysaccharide surface topography was changed from large flakelike structure to smaller chips. FT-IR showed that microwave irradiation did not alter the primary functional groups in the polysaccharide. However, short-chain fatty acid productions of the polysaccharide during in vitro fermentation significantly increased after microwave irradiation. Activities of microbial extracellular enzymes xylanase, arabinofuranosidase, xylosidase, and glucuronidase in fermentation cultures supplemented with microwave irradiation treated polysaccharide were also generally higher than those of untreated polysaccharide. This showed that microwave irradiation could be a promising degradation method for the production of value-added polysaccharides.

Mice (20.0 ± 2.0 g, n = 48 per group) were given 30 days oral administration of polysaccharide from Plantago asiatica L. seeds at the dose of 0.4 g/kg body weight by gavage to investigate the effects of the polysaccharide on mouse colon. Results showed that the concentrations of total short-chain fatty acids (SCFA), acetic, propionic, and n-butyric acids in mouse colonic content of polysaccharide treated group were all significantly higher than that of control group (water) (p < 0.05). In addition, moisture of mouse colonic content of polysaccharide treated group was also notably higher than that of the control group (p < 0.05) indicating the intake of polysaccharide from P. asiatica L. resulted in a stronger water-holding capacity for colonic content throughout the experimental period. Furthermore, a decreased pH (from 7.5 ± 0.1 to 7.2 ± 0.1) was observed in mouse colon of the polysaccharide treated group compared with the control group (pH from 7.5 ± 0.1 to 7.5 ± 0.1). These results suggested that the intake of the polysaccharide from P. asiatica L. might be beneficial for the colon health.

The present study aimed at investigating the mechanism of interactions between calcium and the psyllium polysaccharide. Plantago asiatica L. crude polysaccharide (PLCP) was subjected to ethylenediaminetetraacetic acid (EDTA) to yield calcium-depleted polysaccharide named PLCP-E. There was essentially no difference in the structure between PLCP-E and PLCP. However, PLCP-E exhibited a much lower apparent viscosity compared to that of PLCP. PLCP was treated with sodium hydroxide to deplete ferulic acid. The resultant material was named PLCP-FAS, which also exhibited lower viscosity. Adding Ca(2+) could both increase apparent viscosity of PLCP-E and PLCP-FAS, but only PLCP-E could keep the high viscosity when dialysis was carried out to remove free Ca(2+) in the solution. Thermal analysis showed that the thermal stability of the polysaccharide was reduced after EDTA chelation. Scanning electron microscopy (SEM) analysis showed that PLCP-E was flaky and curly aggregation, while PLCP was mostly filamentous in appearance. The results suggested that there are strong interactions between Ca(2+) and the polysaccharide. The interactions contributed to the high viscosity, weak gelling property, and thermal stability of the polysaccharide.

The aim of this study was to investigate the effects of (-)-epigallocatechin-3-gallate (EGCG) on the induction of apoptosis in hepatocarcinoma cell lines in vitro and further examine the molecular mechanisms of EGCG-induced apoptosis. In the present study, it was observed that EGCG rapidly induced apoptosis in hepatocarcinoma SMMC7721 cells. EGCG-induced apoptosis was in association with the attenuation of mitochondrial transmembrane potentials (Deltapsi(m)), the alteration of Bcl-2 family proteins, the release of cytochrome c from mitochondria into the cytosol, and the activation of caspase-3 and caspase-9. Elevation of intracellular reactive oxygen species (ROS) production was also shown during EGCG-induced apoptosis of hepatocarcinoma SMMC7721 cells. The antioxidant N-acetyl-l-cysteine (NAC) significantly reduced ROS production and EGCG-induced apoptosis, suggesting that ROS plays a key role in EGCG-induced apoptosis in hepatocarcinoma SMMC7721 cells. In summary, EGCG-induced apoptosis through mitochondrial pathways, and ROS affected EGCG-induced apoptosis in hepatocarcinoma SMMC7721 cells.