Zearalenone(ZEA)is widely derived from moldy cereal grain,which has adverse effects on animal reproduction.In particular,pigs are more sensitive to ZEA-induced toxicity than other animals.Iso-rhamnetin has extensive pharmacological activity.However,the role of isorhamnetin in ZEA-induced cytotoxicity remains unclear.This study was designed to investigate the therapeutic effect of iso-rhamnetin on ZEA-induced damage in porcine ovarian granulosa cells and elucidate its molecular mechanism.Two experiments were conducted,where a minimum of 3 biological replicates were used for each treatment.In Exp.1,ovarian granulosa cells were treated with different concentrations of iso-rhamnetin(1,5,10,20 and 30 μmol/L)and ZEA(0,10,30,60,90 and 120 μmol/L)for 24 h.Our results indicated that 60 μmol/L ZEA(half-maximal inhibitory concentration value)and 20 μmol/L isorhamnetin(the most effective concentration against ZEA-induced cytotoxicity)were optimum concentrations.In Exp.2,ovarian granulosa cells were treated with isorhamnetin(20 μmol/L)for 2 h,before treatment with ZEA(60 μmol/L)for 24 h.Apoptosis,endoplasmic reticulum stress,oxidative stress,proliferation and hormone secretion of ovarian granulosa cells were detected.Our findings showed that isorhamnetin suppressed(P＜0.05)ZEA-induced apoptosis by altering mitochondrial membrane potential and apoptosis-related proteins(B-cell lymphoma-2[Bcl-2],Bcl2-associated x[Bax]and cleaved caspase-3[C-Casp3]).Changes in intracellular Ca2+levels and C/EBP homologous protein(CHOP),recombinant activating transcription factor 6(ATF6),glucose regulated protein78 kD(GRP78)indicated that iso-rhamnetin rescued(P＜0.05)ZEA-induced endoplasmic reticulum stress.Furthermore,isorhamnetin prevented(P＜0.05)ZEA-induced oxidative stress via the mitogen-activated protein kinase(P38)signaling pathway.Mechanistically,isorhamnetin stimulated(P＜0.05)the expression of proliferating cell nuclear antigen(PCNA)and cyclin D,thereby increasing the ratio of S phase cells in response to ZEA-induced apoptosis via phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)signaling pathway.Isorhamnetin also recovered(P＜0.05)ZEA-induced steroidogenesis disorder by regulating steroidogenic enzyme gene and proteins(follicle-stimulating hormone receptor[FSHR]and cytochrome P450 family 19 subfamily a member 1[CYP19A1]).Collectively,these findings show that isorhamnetin protects ovarian granulosa cells from ZEA-induced damage,which promotes proliferation,alleviates apoptosis,endo-plasmic reticulum stress,oxidative stress,and steroidogenesis disorder.

Xiaoya Li;Huali Chen;Zelin Zhang;Jiaxin Duan;Rongmao Hua;Xiaodi Li;Li Yang;Jianyong Cheng;Qingwang Li

College of Animal Science and Technology,Northwest A&F University,Yangling,712100,China;School of Life Science and Engineering,Southwest University of Science and Technology,Mianyang 621000,China;College of Basic Medicine,Hebei University of Chinese Medicine,Shijiazhuang,050000,China

动物营养（英文）

[1]Xiaoya Li;Huali Chen;Zelin Zhang;Jiaxin Duan;Rongmao Hua;Xiaodi Li;Li Yang;Jianyong Cheng;Qingwang Li-.Isorhamnetin protects zearalenone-induced damage via the PI3K/Akt signaling pathway in porcine ovarian granulosa cells)[J].动物营养（英文）,2022(04):381-390

rhamnetin,Ca2+levels,protein78

Isorhamnetin,protects,zearalenone,induced,damage,PI3K,Akt,signaling,pathway,porcine,ovarian,granulosa,cells,Zearalenone,ZEA,widely,derived,from,moldy,cereal,grain,which,adverse,effects,reproduction,In,particular,pigs,are,sensitive,than,other,animals,extensive,pharmacological,activity,However,role,isorhamnetin,cytotoxicity,remains,unclear,This,study,was,designed,investigate,therapeutic,elucidate,its,molecular,mechanism,Two,experiments,were,conducted,where,minimum,biological,replicates,used,each,treatment,Exp,treated,different,concentrations,Our,results,indicated,that,half,maximal,inhibitory,value,most,effective,against,optimum,before,Apoptosis,endoplasmic,reticulum,stress,oxidative,proliferation,hormone,secretion,detected,findings,showed,suppressed,apoptosis,altering,mitochondrial,membrane,potential,related,proteins,lymphoma,Bcl2,associated,Bax,cleaved,caspase,Casp3,Changes,intracellular,EBP,homologous,CHOP,recombinant,activating,transcription,ATF6,glucose,regulated,kD,GRP78,rescued,Furthermore,prevented,mitogen,activated,kinase,P38,Mechanistically,stimulated,expression,proliferating,nuclear,antigen,PCNA,cyclin,thereby,increasing,phase,response,phosphatidylinositol,also,recovered,steroidogenesis,disorder,regulating,steroidogenic,enzyme,follicle,stimulating,receptor,FSHR,cytochrome,P450,subfamily,member,CYP19A1,Collectively,these,promotes,alleviates

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