1.Resveratrol protects against oxidative stress damage during sperm cryopreservation by mediating the NRF2 signaling pathway
Lei XIN ; Yutong ZHANG ; Liting LIU ; Jing TAN ; Jingjing ZHANG ; Zongliu DUAN ; Yang GAO ; Jun YANG ; Biao YU ; Qunshan SHEN ; Xiaojin HE
Chinese Journal of Reproduction and Contraception 2025;45(10):1038-1046
Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.
2.Resveratrol protects against oxidative stress damage during sperm cryopreservation by mediating the NRF2 signaling pathway
Lei XIN ; Yutong ZHANG ; Liting LIU ; Jing TAN ; Jingjing ZHANG ; Zongliu DUAN ; Yang GAO ; Jun YANG ; Biao YU ; Qunshan SHEN ; Xiaojin HE
Chinese Journal of Reproduction and Contraception 2025;45(10):1038-1046
Objective:To investigate the protective effects of resveratrol (RSV) on human sperm cryopreservation and explore its underlying protective mechanisms.Methods:A total of 165 normal fresh semen samples were collected from the Reproductive Medicine Center, Department of Obstetrics and Gynecology and Human Sperm Bank of the First Affiliated Hospital of Anhui Medical University between December 2022 and December 2024. Among them, 65 samples were used to obtain semen parameters before and after conventional freezing. Each sample of the other 104 samples was mixed at a 2∶1 volume ratio with cryoprotectant containing 0, 10 -?, 10 -?, or 10 -? mol/L RSV, followed by cryopreservation in liquid nitrogen for 24 h. Post-thaw assessments included routine sperm parameters, sperm DNA fragmentation index (DFI) evaluated by sperm chromatin dispersion assay, reactive oxygen species (ROS) levels measured via flow cytometry, RSV and nuclear factor erythroid 2-related factor 2 (NRF2) interactions examined by molecular docking and cellular thermal shift assay (CETSA), NRF2 protein contents analyzed by immunofluorescence and Western blotting, mRNA levels of NRF2 and downstream antioxidant proteins Heme Oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1) quantified by qRT-PCR and effects of NRF2 inhibitor ML385 on sperm parameters. Results:Compared with fresh samples, conventional cryopreservation significantly reduced sperm motility (all P<0.001). The addition of 10 -? mol/L RSV significantly improved the percentage of forward motile sperm after freezing (26.98%±8.98% vs. 19.61%±8.03%, P<0.001) while reducing DFI (9.84%±3.81% vs. 15.06%±4.22%, P<0.001) and ROS levels ( P<0.001) compared with the post-freezing group without the addition of RSV. Both molecular docking analysis and CETSA confirmed that RSV interacted with NRF2. Notably, sperm cryopreserved with 10 -? mol/L RSV exhibited significantly higher contents of NRF2 and its downstream effectors HO-1 and NQO1 compared with the post-freezing group without the addition of RSV (all P<0.001). This protective effect was markedly attenuated by co-treatment with the NRF2 inhibitor ML385, as evidenced by a significant decline in sperm motility ( P<0.001). Conclusion:RSV exerts cryoprotective effects likely through NRF2-mediated antioxidant pathways, reducing oxidative stress and enhancing post-thaw sperm quality.
3. Relationship between bile acid metabolism and chronic non - cholestatic liver disease
Liting YAN ; Junfeng LI ; Zhongping DUAN ; Qingfeng CHEN
Chinese Journal of Hepatology 2018;26(2):151-154
Bile acid is a general term for a large class of cholic acid in bile and exerts its unique physiological functions by binding the relevant receptors and bile acid transporters. Bile acids not only promote the absorption of nutrients in the human body, but also as an important signaling molecule in the regulation of inflammatory processes and liver regeneration. Several studies have found that bile acid metabolism is involved in the occurrence and development of chronic non-cholestatic liver diseases. In this article, the latest research results are mentioned, and the relationship between nuclear receptors, membrane receptors, and bile acid transporters and chronic non-cholestatic liver disease that play a key role in the bile acid metabolism were emphatically reviewed.
4.Effect of chronic high-fat diet on predation behavior in rats
Wenhao CHEN ; Yan JIA ; Shuhui CAO ; Yaru CHEN ; Liting DUAN ; Changqi LI
Journal of Central South University(Medical Sciences) 2014;(12):1266-1270
Objective: To observe the eff ect and mechanism of chronic high-fat diet on predation behavior in rats. Methods: Ten female SD rats with 4-week-old were randomly divided into a normal control group (NC group,n=5) and a chronic high-fat diet group (HF group,n=5). The rats in the NC group received the regular diet while rats in the HF group were fed with high-fat diet. Fitf een weeks later, the predation behavior of rats was evaluated by open if eld test and food foraging tests. At the end of experiments, the rats were killed and brain tissues were collected for evaluation of c-Fos protein expression in anterior cingulate cortex by immunohistochemical assay. Results: hT e predation behavior of rats in the HF group was signiif cantly impaired in the competitive or non-competitive food foraging test compared with the control rats (P<0.001). hT e c-fos protein expression in anterior cingulate cortex of rats from the HF group was signiif cantly decreased (P<0.001). Conclusion: Long time high-fat diet can aff ect the predation behavior of rats, which is related todysfunction of neuron in anterior cingulate cortex.

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