Cold stress aggravates reproductive dysfunction in rats with polycystic ovary syndrome
Objective To investigate the effects of cold stress stimulation on reproductive function of rats with polycystic ovary syndrome(PCOS)and the underlying mechanism.Methods Rats were randomly divided into control group,model group,cold stress group,inhibitor group(HSP90 inhibitor 50mg/kg)and combination group(cold stress combined with HSP90 inhibitor),with 10 in each group.Serum sex hormone and oxidative stress levels were detected by enzyme-linked immunosorbent assay(ELISA).Ovarian morphology was observed by hematoxylin and eosin(H&E)staining.Apoptosis of ovarian cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).Protein expressions of heat shock protein 90(HSP90),nuclear factor-kappaB(NF-kappaB)P65,inhibitor kB(IkB),transforming growth factor-beta-activated kinase 1(TAK1)and RNA immunoprecipitation(RIP)were detected by Western blot.Results Follicle-stimulating hormone(FSH,[7.21±1.54]mU/mL vs[7.25±1.51]mU/mL vs[16.37±2.14]mU/mL,P<0.05)in the model group and combination group was significantly lower than that of the control group.Luteinizing hormone(LH,[20.34±2.81]nmoL/L vs[20.28±2.85]nmoL/L vs[9.32±1.46]nmoL/L,P<0.05)and testosterone(T,[1.58±0.26]nmoL/L vs[1.56±0.27]nmoL/L vs[1.23±0.17]nmoL/L,P<0.05)in the model group and combination group were significantly higher than those of the control group.Compared with those of the model group,the inhibitor group showed significantly reduced FSH([7.21±1.54]mU/mL vs[6.28±1.24]mU/mL,P<0.05),and significantly increased LH([20.34±2.81]nmoL/L vs[22.08±2.73]nmoL/L,P<0.05)and T([1.58±0.26]nmoL/L vs[1.64±0.25]nmoL/L,P<0.05).Compared with those of the inhibitor group,the cold stress group showed significantly reduced FSH([6.28±1.24]mU/mL vs[5.34±0.67]mU/mL,P<0.05),and significantly increased LH([22.08±2.73]nmoL/L vs[23.17±2.95]nmoL/L,P<0.05)and T([1.64±0.25]nmoL/L vs[1.79±0.31]nmoL/L,P<0.05).Compared with those of the control group,the model group and the combination group showed significantly reduced superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and significantly increased malondialdehyde(MDA)(P<0.05).Compared with those of the model group,the inhibitor group showed significantly increased SOD and GSH-Px,and significantly decreased MDA(P<0.05).Compared with those of the inhibitor group,the cold stress group showed significantly reduced SOD and GSH-Px,and significantly increased MDA(P<0.05).Rat ovarian tissue in the control group was normal.Rat ovarian tissue in the model group and combination group showed reduced cortical corpus luteum and follicles,cystic expansion of follicles,and atresia.Rat ovarian tissue in the cold stress group and inhibitor group mainly showed a large number of small antral follicles,cystic follicles and enlarged theca cell layers,which were much severer in the cold stress group.The cell apoptotic rate of the control group,model group,inhibitor group,cold stress group and combination group was(5.62±0.58)%,(41.38±3.92)%,(52.67±4.22)%,(63.48±5.71)%and(40.62±4.28)%,respectively.The cell apoptotic rate in the ovarian tissue of the control group was significantly lower than that of the model group and combination group(F=168.200,P<0.001).The cell apoptotic rate in the ovarian tissue of the inhibitor group was significantly higher than that of the model group(t=6.213,P<0.001).The cell apoptotic rate in the cold stress group was significantly higher than that of the inhibitor group(t=4.815,P<0.001).Compared with those of the control group,the expression levels of HSP90,NF-κB P65,IkB,TAK-1 and RIP in rat ovarian tissue of the model group were significantly upregulated(P<0.05).Compared with those of the model group,the expression levels of HSP90,NF-κB P65,IkB,TAK-1 and RIP in rat ovarian tissue of the cold stress group and control group were significantly upregulated(P<0.05).There was a positive correlation between the expressions of HSP90 and PI3K in rat ovarian tissue after cold stress stimulation(r = 0.581,P<0.001).Conclusion Cold stress aggravates reproductive dysfunction in PCOS rats.
heat shock protein 90(HSP90)-related signalcold stress responsepolycystic ovary syndrome(PCOS)ratsreproductive function
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维普
