Developing natural marine products for treating liver diseases

INTRODUCTION

Liver diseases are rapidly emerging as global health priorities.With increasingly urbanized lifestyles and dietary changes involving high caloric contents,the overall prevalence of non-alcoholic fatty liver disease(NAFLD)has increased dramatically.Non-alcoholic steatohepatitis(NASH)has become one of the leading causes of liver transplantation in the United States[1].NAFLD is associated with metabolic syndrome and the development of cardiovascular and kidney diseases.Alcoholic liver disease(ALD)is caused by heavy alcohol intake.Almost 50% of cirrhosis-related deaths are due to excessive alcohol consumption[2].Hepatitis B virus(HBV)infection is the most common cause of chronic hepatitis worldwide and remains the primary cause of cirrhosis and hepatocellular carcinoma(HCC)in most Asian nations.Hepatitis C virus(HCV)has infected 71 million people worldwide[3].HCC is one of the most common malignant tumors worldwide.HBV,HCV,NAFLD,and ALD are significant risk factors for HCC.Drug-induced liver injury(DILI)is an important cause of acute liver failure.All chronic liver diseases can lead to liver cirrhosis and decompensation,thus requiring effective and cost-efficient treatments.

The ocean accounts for 70% of the Earth’s surface area[4].Marine organisms are known for their ability to produce large amounts of bioactive compounds,whose biological activities could interfere with the pathogenesis of many diseases.Interest in marine organisms as a source of health-promoting agents has increased in recent decades.Marine organisms are classified as marine plants(,seaweeds and mangroves),marine animals(,sponges,corals,shellfish,krill,and ascidians),and marine microorganisms,according to their biological characteristics.They have been found to be rich sources of bioactive compounds with anti-inflammatory,antioxidant,antibacterial,antiviral,anti-tumor,and lipidlowering activities.This review discusses current applications of bioactive marine compounds in studying liver diseases(briefly summarized in Table 1).

NAFLD

NAFLD is a spectrum of common liver diseases and currently is responsible for a global disease epidemic with an estimated worldwide prevalence of 25%[5].The highest rates are reported in South America and the Middle East,followed by Asia,the USA,and Europe.NAFLD includes a range of diseases ranging from fatty liver to NASH,liver fibrosis,cirrhosis,and liver cancer.Insulin resistance,lipotoxicity,mitochondrial dysfunction,oxidative stress,intestinal microbiome disorders,and genetic and epigenetic factors are related to NAFLD pathogenesis[6].Data from several studies have shown improvements in patients with NASH after treatment with vitamin E,liraglutide,statins,glitazones,and pioglitazone[7].However,no special therapeutic medications have been approved by the Federal Drug Administration(FDA).In the absence of effective pharmacological agents for NAFLD,lifestyle interventions such as increased exercise and energy restriction,lowering hepatic lipid levels,and increasing insulin sensitivity are important measures.Several studies of bioactive marine substances have provided new therapeutic prospects for NAFLD.

Fish oil contains a variety of n-3 long-chain polyunsaturated fatty acids(PUFAs),including docosahexaenoic acid(DHA)and eicosapentaenoic acid(EPA),which can activate the peroxisome proliferator receptor(PPAR)and downregulate the expression of sterol regulatory element binding protein 1c(SREBP-1c)and carbohydrate response element-binding protein(ChREBP).N-3 PUFA can not only protect against dyslipidemia,insulin resistance,and obesity,but also has anti-inflammatory and antioxidant properties.Previous findings have shown that n-3 PUFA supplementation can prevent NAFLD[8].Functional lipids from the starfishoil,such as n-3 PUFA and carotenoids(which have antioxidant activities and can preserve insulin sensitivity),dose-dependently decreased liver lipid accumulation and improved liver steatosis in C57BL/6N mice fed a high-fat diet(HFD)[9].Krill represent a rich source of protein with essential amino acids and minerals.In fish oils,EPA and DHA are present in the form of triacylglycerol(TAG),whereas they are present as phospholipids(PLs)in krill,which has stronger anti-inflammatory and insulin hypersensitivity properties.A krill phospholipid-protein complex(PPC)fromcan reduce hepatic lipogenesis in rats,which is associated with an increased total antioxidant capacity.

Sea cucumber sulfated polysaccharide(SCSP)inhibits the expression of the main regulatory mediator of liver lipid genesis,SREBP-1c,which leads to inhibited hepatic triglyceride synthesis.SCSP also significantly increases PPARα expression,thus promoting the β oxidation of fatty acids.SCSP is barely absorbed in the gut,which leads to modulation of the gut microbiota.Therefore,SCSP may have unique effects on NAFLD and other pathological liver diseases[10,11].

Carotenoids are natural pigments with strong antioxidant activities.Their benefits in treating liver diseases and related complications have been widely reported.Astaxanthin(AST)is an important xanthophyll carotenoid,which is mainly derived from marine organisms and algae.Its antioxidant effect is 10 times that of β-carotene and 100 times that of vitamin E.In addition to its strong antioxidant properties,AST can also regulate various signaling pathways,such as reducing the activities of JNK and ERK-1 to lower liver insulin resistance,inhibiting PPAR-γ expression to reduce liver fat synthesis,

downregulating TGF-β1/Smad3 expression to inhibit hepatic stellate cell(HSC)activation and liver fibrosis,and inhibiting the JAK/signal transducer and activator of transcription 3(STAT3)and Wnt/βcatenin signaling pathways to exert antitumor effects.Therefore,AST plays significant roles in preventing and treating NAFLD,liver fibrosis,HCC,DILI,and ALD[12].Siphonaxanthin(SPX)is a carotenoid derived from green marine algae that can significantly inhibit liver X receptor α(LXRα)activity and downregulate the expression of SREBP-1c and several related genes to inhibit liver adipogenesis[13].

Data from several studies showed that various bioactive components from brown algae can alleviate liver steatosis to a certain extent,especially fucoidan.Fucoidan is a sulfated polysaccharide extracted from brown marine algae that can regulate the ROS/JNK/Akt signaling pathways,reduce insulin resistance,inhibit sugar transport,regulate lipid metabolism and the gut microbiota,and reduce liver steatosis[14].Some other bioactive compounds from brown algae,such as indole-4-carboxaldehyde,unsaturated alginate oligosaccharides(UAOS),and diphlorethohydroxycarmalol(DPHC),can inhibit inflammation and lipid metabolism,but further clinical trials are needed to confirm their efficacies against NAFLD.Ananimal study showed that the red algaecan upregulate PPAR α expression,thereby activating fatty acid β oxidation and inhibiting lipid synthesis to improve liver steatosis[15].

Spirulina are cyanobacteria capable of photosynthesis,which implies that they are rich in antioxidants.PUFAs in a 95% ethanol extract of the microalga(SPL95)can regulate the gut microbiota,and reduce lipid synthesis and liver fat in rats fed an HFD by upregulating AMPK-α and downregulating members of the SREBP-1c-signaling pathway[16].

Asperlin is a natural fungal product isolated from the marine-derived fungusLZD-44-03.Asperlin improved lipid metabolism,ameliorated liver steatosis,and modulated the gut microbiota in mice fed an HFD[17].

is a marine animal that synthesizes bioactive components with anti-tumor,antibacterial,and antiviral effects.Previous data showed that the bioactive compounds of ascidians can inhibit HBV DNA replication and have potential therapeutic value against chronic HBV infection[36].Yamashita[37]also showed that metachromin A,a merosesquiterpene isolated from the marine sponge,can inhibit HBV production by impairing viral promoter activity.

In recent years,increasing attention has been paid to the anti-tumor activities of cockles.Some data have shown that polypeptides derived from cockles can exert anti-tumor activity by inhibiting the MAPK signaling pathway[55].Guo[56]found that the ASP-3 protein isolated frominhibited HepG2 cell proliferation.

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Meretrix meretrix oligopeptides(MMOs)are substances with important medicinal value that are extracted from shellfish.anddata have shown that MMOs can reduce oxidative stress,improve mitochondrial dysfunction,and inhibit the activation of cell death-related pathways,thus exerting protective effects against NAFLD[18].

is one of the largest jellyfish species.The venom(NnV)contains highly selective dual inhibitors of the Akt and mTOR signaling pathways,which induce cytotoxicity and apoptosis in HepG2 cells,but not normal cells[58].Yet,NnV can inhibit the metastasis and invasion of HepG2 cells by inhibiting the epithelial–mesenchymal transition[59].

ALD

Alcohol abuse is the seventh leading risk factor for death globally,and the liver is the main organ involved in alcohol metabolism.Excessive alcohol intake can damage liver cells and cause ALD[19].The risk is increased in people who have heavy alcohol use(>3 drinks per day in men and >2 drinks in women)for >5 years.Heavy drinking increases intestinal permeability and the influx of lipopolysaccharide(LPS)to the liver,activates Kupffer cells,and leads to high Toll-like receptor 4(TLR4)expression,which in turn releases large amounts of ROS and tumor necrosis factor(TNF-α)or other inflammatory factors,leading to liver toxicity.Drinking can also reduce the PL levels in liver cell membranes.Environmental factors andandgene mutations can also induce ALD progression[20].Current treatments for ALD depend on ensuring lasting alcohol abstinence,and the treatment strategies beyond alcohol abstinence are largely those used for complications of cirrhosis,such as controlling ascites,treating and preventing hepatic encephalopathy recurrence and variceal bleeding,and monitoring for hepatocellular cancer[21].

Recent data have shown the effects of natural extracts on preventing and/or lessening alcoholic liver injury.AST may prevent ALD progression through pathways related to chemokine signaling,NOD-like receptor signaling,and TLR signaling[22].

Aplysin was extracted from the red algaand exerts a potent hepatoprotective effect against ALD by enhancing the antioxidant defense system,alleviating oxidative damage,and regulating apoptosis-related gene expression[23].

Fucoxanthin(Fx)is a red-orange carotenoid extracted from marine seaweed that has strong antiobesity,anti-inflammatory,and anti-cancer activities.data indicated that Fx attenuated alcoholinduced oxidative lesions and inflammatory responses by activating the nuclear factor erythrocyte-2-related factor 2(Nrf2)-mediated signaling pathway and downregulating the expression of members of the TLR4-mediated nuclear factor-kappa B(NF-κB)signaling pathway,respectively.

The woman was still raging, and did not seem likely to stop, when her husband, who by this time had forgotten all about her, drew out the chest from under his cloak, and opened it

Marine collagen peptides(MCPs)are derived from the skin of chum salmon()by enzymatic hydrolysis;MCPs can protect against early alcoholic liver injury in rats,based on their antioxidative activities and improvements in terms of lipid metabolism[24].

Krill()-derived peptides are renowned for their antioxidant activities,and peptide fractions from krill protein hydrolysates protect against alcohol-induced oxidative damage in BALB/c mice.This hepatoprotective effect might be attributed to activation of the Nrf2/HO-1 pathway.

HCV INFECTION

HCV results in an infectious liver disease,with multiple genotypes.HCV infection can lead to steatosis,liver cirrhosis,and HCC.Approximately 3% of the world’s population are infected with HCV[25].The most effective HCV treatment regimen depends on the genotype of the predominant viral strain in infected patients.Overall,there are 11 HCV genotypes,with genotypes 1–6 being the most common[26].Traditional therapy involves treatment with a combination of pegylated interferon alpha and ribavirin.The current FDA-approved direct acting antivirals are commonly used in combinations as pangenotypic to effectively inhibit HCV replication with minimal side effects.However,the occurrence of resistance(either natural or after failure)and drug-drug interactions can limit treatment effectiveness.Natural HCV inhibitors still need to be investigated.

Harzianoic acids A and B,isolated from the sponge-relatedfungus,inhibit viral activity by reducing RNA levels[27].Total extract and derived fractions from red seasponges exhibited inhibitory potential against HCV NS3 helicase and protease.Among.-derived phytochemicals,nakinadine B and 3,4-dihydro-6-hydroxymanzamine A were noted as promising anti-HCV drug candidates,warranting future clinical investigation[28].Fucoidan extracted from the marine alga()Tokida dose-dependently inhibited an HCV replicon system,suggesting that fucoidan may be a useful food additive with antiviral activity for treating chronic liver diseases[29].

Lobohedleolide isolated from the formosan soft coral,significantly reduced HCV replication by suppressing cyclooxygenase-2(COX-2)expression[30].

Meanwhile the mother dove sat on the edge of her nest, crying bitterly, when a heron, who was flying slowly past the rock, was filled with pity for her, and stopped to ask, What is the matter, you poor dove? And the dove answered, A jackal came by, and asked me to give him one of my little ones, and said that if I refused he would jump on my nest and eat us all up

"The real challenge of deep learning is that it's not modeling, necessarily, the world around it. It's modeling the data it's getting," Fernandez said. "And that modeling often includes bias and problematic correlations. It can include nonsensical correlations. And all those things can find [their] way into the behavior of the system."

Various active components isolated from the soft coral,such as 11-epi-sinulariolide acetate and sinulariolide,could inhibit HCC cell migration and invasion by reducing matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),and urokinase-type plasminogen activator(uPA)expression[53].Sinularin induced DNA damage,G2/M phase arrest,and apoptosis[54].

Fucoidan fromwas found to protect against alcohol-induced liver damage in mice.The associated mechanism potentially involved suppressing hepatic production of inflammatory cytokines,such as TGF-β1,COX-2,and NO,and enhancing oxidant defense systems by activating the HO-1 pathway.

DILI

The main elimination mechanisms of exogenous drugs involve the liver,kidney,and bile.Sixty percent of drugs are metabolized by the liver.DILI is a type of liver disease caused by drugs and their metabolites.Severe cases are life-threatening.The incidence of clinically significant DILI varies from country to country.Despite its rarity(<1%,as determined with most patient series),it has been found to be the most common cause of acute liver failure in both Europe and the United States[31].The most important initial step in terms of managing suspected DILI is to discontinue the implicated agent,as ongoing or even worsening injury can occur despite withdrawal of the causative agent.Drugs presently used to treat DILI are mainly those that protect liver cells,scavenge free radicals,inhibit oxidation,stabilize cell membranes,promote detoxification,lower enzymes,and promote immune regulation,including ursodeoxycholic acid,N-acetylcysteine,various steroids,and glutathione,among others.However,there is still a lack of drugs that treat DILI specifically.

Fucoidan displayed a hepatoprotective effect on acetaminophen overdose-induced liver toxicity,based on the suppression of CYP2E1,one of the enzymes that metabolizes acetaminophen.Fucoidan also exerts anti-oxidant,anti-apoptotic,and anti-inflammatory activities by increasing the production and expression of glutathione,superoxide dismutase,glutathione peroxidase,and Bcl-2,but decreasing the expression of Bax,cleaved caspase-3,and inflammatory mediators,including TNF-α,IL-1β,and iNOS[32].An ethanolic extract of(red algae)was found to possess antioxidant,antitumor,and antimicrobial activities and to exhibit hepatoprotective activity against CCl-induced toxicity in rats[33].

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Fish oil can reduce liver damage caused by lipopolysaccharides,cisplatin,and acetaminophen by inhibiting TLR4 and nucleotide-binding oligomerization domain protein signaling pathways,and their antioxidant properties[34,35].

We were stopped and waiting for another rider to be helped when my young student reached out and touched my hair. My hand was on his leg, so I knew he was steady, even though my eyes weren t on him. I looked around and knew he was trying to tell me something. The horse stood motionless, as if he knew his movement could distract6 or confuse his rider. What? I asked. It was unusual for him to reach out and touch, to even control his hands enough to do so. He reached out again and stroked my hair, as he sometimes did to the horse s mane on good days.

OTHER LIVER DISEASES

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The farnesoid X receptor(FXR)can mediate bile acid secretion,and theonellasterol(isolated from the marine sponge)is a highly selective FXR antagonist that can protect against liver injury in cholestasis[38].

LIVER FIBROSIS

Liver fibrosis is a scar-repair process that occurs after liver injury caused by various factors.It is characterized by liver myofibroblast cell(MFC)activation and excessive accumulation of extracellular matrix(ECM)proteins,and is pathologically characterized by the formation of regenerative nodules of hepatocytes,which can lead to cirrhosis and liver failure.Chronic HBV and HCV infections,alcoholic steatohepatitis,and NASH are the main causes of chronic progressive liver disease,leading to the onset of liver cirrhosis and decompensation.During chronic liver injury,silent HSCs are activated to become highly proliferative MFCs at the cellular level,resulting in α-smooth muscle actin(α-SMA)expression and excessive production of type I and type III collagen,as well as other scar tissue components.At the molecular level,pro-fibrotic factors,including transforming growth factor-β1(TGF-β1),platelet-derived growth factor,and connective tissue growth factor;multiple signaling pathways such as the TLR4,and damage due to reactive oxygen species(ROS)play key roles in this process[39].

Owing to the unique chemical properties of sulfur atoms,sulfur-containing compounds are powerful antioxidants that exhibit promising activities for treating liver fibrosis.For example,ovothiol A(a sulfur-containing molecule)isolated from sea urchin eggs was found to have an anti-fibrotic effect on mice with carbon tetrachloride(CCl)-induced liver fibrosis.This anti-fibrotic effect may be related to reduced expression of mediators involved in the progression of liver fibrosis,such as TGF-β,α-SMA,and tissue inhibitor of metalloproteinase(TIMP-1)[40].

In addition,Nakazato[41]found that fucoidan fromTokida reduced N-nitrosodiethylamine-induced liver fibrosis.It exerted an anti-fibrotic effect by downregulating TGF-β1 and CXCL12 expression and reducing lipid peroxidation.

Spirulina liquid extract(SLE),a patented water extract of,protects against hepatic fibrosis by inhibiting inflammation,oxidative stress,and whole-body insulin resistance in a mouse model of Western diet-induced NASH[42].Astaxanthin(AST)from starfish and algae exerts anti-fibrotic effects through the TGF-β1/Smad3 signaling pathway in hepatic stellate cells[43].

HCC

HCC is the most common primary liver cancer,accounting for approximately 75%-85% of such cases[44],and the third leading cause of cancer-related mortality worldwide.HCC has a high fatality rate,with a 5-year survival rate of only 30%-40%[45].An estimated 70%-90% of HCC cases arise in the setting of cirrhosis[46].HCC treatment mainly includes liver resection,liver transplantation,radiofrequency or microwave ablation,radiotherapy,and chemotherapy.Sorafenib,a multi-kinase VEGF inhibitor,is the most widely used systemic chemotherapeutic drug approved as a first-line agent for unresectable or advanced HCC.Other small-molecule inhibitors such as sunitinib,brivanib,and erlotinib have been studied for their efficacy in treating advanced HCC[47].However,these anti-tumor drugs still have disadvantages related to drug resistance,poor efficacy,and large side effects.In recent decades,investigators have become committed to researching natural products as new anti-tumor drugs.

Sponges host diverse microbial communities,such as fungi,bacteria,and microalgae,and are rich in bioactive peptides that are important candidates for drug development.Data from many studies have shown that various sponge metabolites have anti-tumor activities in liver cancer cells.Ceratamine A and B isolates fromsponges can behave as antimitotic agents by disrupting microtubule dynamics[48].Similarly,(+)terrein,isolated from the marine spongestrain PF-26,can inhibit human hepatoma Bel7402 proliferation by blocking the expression of genes related to cell cycle progression and changing the cell morphology[49].Crambescidin-816,purified from the sponge,exerted an anti-tumor effect by inhibiting cell–cell adhesion,interfering with tight junction formation and cell–matrix adhesion,negatively affecting focal adhesions,and altering cytoskeletal dynamics[50].Actinomycetales isolated from marine sponges of threegenera collected in Portuguese waters showed anti-cancer activity against HCC cell line[51].,the marine-derived steroid methyl spongoate(MESP)molecule from Sanya soft coralpotently induced apoptosis by activating a proapoptotic caspase cascade and relieving suppression of antiapoptotic STAT3 signaling[52].

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Chitosan oligosaccharide(COS),a natural polysaccharide hydrolyzed from shrimp shell chitosan,has attracted extensive attention because of its potential use in various promising biomedical applications,including those related to anti-oxidation,anti-inflammation,immune stimulation,and antihypertension.An enzymatically digested product of COS,known as COS23,can reduce hepatotoxic lipid levels,inhibit the expression of FFA synthesis-related genes and inflammatory-related genes,regulate the gut microbiota,and up-regulate the expression of tight junction-related genes to improve intestinal barrier dysfunction,thereby improving diet-induced NAFLD.

Late one evening, as my husband snoozed in front of the television and I was catching7 up on my e-mail, an instant message appeared on the screen. It was Margo, my oldest step-daughter, also up late and sitting in front of her computer five hours away. As we had done in the past, we sent several messages back and forth8, exchanging the latest news. When we would chat like that, she wouldn t necessarily know if it was me or her dad on the other end of the keyboard--that is unless she asked. That night she didn t ask and I didn t identify myself either. After hearing the latest volleyball scores, the details about an upcoming dance at her school, and a history project that was in the works, I commented that it was late and I should get to sleep. Her return message read, Okay, talk to you later! Love you!

Marine microorganisms can produce various metabolites with unique structures and pharmacological activities,such as SZ-685C[57],which is a natural biologically active substance isolated from secondary metabolites of marine mangrove endophytic fungus number 1403 and could induce apoptosis through the Akt/FOXO pathway.EPS11,a bacterial polysaccharide extracted from11,can inhibit the growth and metastasis of HCV-related liver cancer cells(Huh7.5 cells)by blocking their adhesion and destroying the formation of filamentous structures.

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Fucoidan extracted from the brown seaweedinduces apoptosis in human HCC SMMC-7721 cells by increasing ROS production and inducing mitochondrial oxidative damage,mitochondrial membrane potential depolarization,and caspase activation.Fucoidan can also reduce lymphangiogenesis and tumor lymphatic(by suppressing HIF-1α/VEGF-C signaling),and then attenuate the PI3K/Akt/mTOR signaling pathways.Phycobiliproteins are components of red algae that include phycoerythrin,phycoerycyanin,phycocyanin,and allophycocyanin,which can have antioxidative,anti-viral,anti-tumor,immunity-enhancing,and anti-inflammatory effects[60].Park[61]demonstrated that dietary RPE could modulate the gut microbiota of H22 HCC cell-bearing mice.experiments have revealed that COS showed significant antitumor activity against HepG2 tumor cells.

Ovothiol A,isolated fromoocytes,can inhibit HepG2 cell proliferation by activating an autophagic process.

CONCLUSION

Treatments for chronic liver diseases,,etiological treatment,protective treatment for liver injury,antifibrotic treatment,and treatment of decompensated complications of liver cirrhosis,are all effective and of great significance in preventing the progression of liver disease,maintaining liver function,reducing complications of portal hypertension,and preventing liver cancer.In recent years,intense research of marine resources has brought to light new prospects for developing marine-based drugs.Marine-derived bioactive compounds show great potential in health products and medicine.With the discovery of bioactive marine compounds and in-depth discussions of their therapeutic mechanism,their applications will continue to expand,and ultimately benefit more patients and humans.However,most of the information discussed above has only been demonstratedor in animal studies.The effects of these compounds in humans have yet to be fully characterized.Further research,including clinical trials,must be carried out before such marine compounds can be applied therapeutically.

FOOTNOTES

Qian W drafted the manuscript;Guo JS conceptualized and revised the manuscript;all authors have read and approved the final manuscript.

the National Natural Science Foundation of China,No.91129705,No.81070340,and No.30570825.

The authors declare no conflicts of interest for this article.

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China

Qian Wei 0000-0003-2702-0207;Jin-Sheng Guo 0000-0002-9980-8725.

American Association for the Study of Liver Diseases.

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Yan JP