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Effects of Mitochondrial Uncoupling Protein 2 Inhibition by Genipin in Human Cumulus Cells.

Ge H, Zhang F, Shan D, Chen H, Wang X, Ling C, Xi H, Huang J, Zhu C, Lv J - Biomed Res Int (2015)

Bottom Line: There were no significant differences in ATP and mitochondrial membrane potential levels in cumulus cells from UCP2 inhibiting groups as compared with the control.However, the ratio of reduced GSH to GSSG significantly declined after treatment with Genipin.But there was no significant difference in estradiol concentrations.

View Article: PubMed Central - PubMed

Affiliation: Reproductive Health Center, Second Affiliated Hospital of Wenzhou Medical University, 109 Xueyuan Road, Wenzhou, Zhejiang 325000, China.

ABSTRACT
UCP2 plays a physiological role by regulating mitochondrial biogenesis, maintaining energy balance, ROS elimination, and regulating cellular autophagy in numerous tissues. But the exact roles of UCP2 in cumulus cells are still not clear. Genipin, a special UCP2 inhibitor, was added into the cultural medium to explore the roles of UCP2 in human cumulus cells. There were no significant differences in ATP and mitochondrial membrane potential levels in cumulus cells from UCP2 inhibiting groups as compared with the control. The levels of ROS and Mn-SOD were markedly elevated after UCP2 inhibited Genipin. However, the ratio of reduced GSH to GSSG significantly declined after treatment with Genipin. UCP2 inhibition by Genipin also resulted in obvious increase in the active caspase-3, which accompanied the decline of caspase-3 mRNA. The level of progesterone in culture medium declined obviously after Genipin treatment. But there was no significant difference in estradiol concentrations. This study indicated that UCP2 is expressed in human cumulus cells and plays important roles on mediate ROS production, apoptotic process, and steroidogenesis, suggesting UCP2 may be involved in regulation of follicle development and oocyte maturation and quality.

No MeSH data available.


Effects of UCP2 inhibited by Genipin on CCs apoptosis. (a) The caspase-3 protein was quantified by Western blotting. The proactive caspase-3 declined obviously, and the active caspase-3 markedly increased after UCP2 inhibition. (b) Caspase-3 gene was quantified by RT-PCR, which markedly increase after UCP2 inhibition. Con means control, asterisks indicate significant difference between control and treatment group at P < 0.05.
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fig3: Effects of UCP2 inhibited by Genipin on CCs apoptosis. (a) The caspase-3 protein was quantified by Western blotting. The proactive caspase-3 declined obviously, and the active caspase-3 markedly increased after UCP2 inhibition. (b) Caspase-3 gene was quantified by RT-PCR, which markedly increase after UCP2 inhibition. Con means control, asterisks indicate significant difference between control and treatment group at P < 0.05.

Mentions: To determine effects of UCP2 on cellular redox status in CCs, Mn-SOD activity and the levels of the ROS, ratio of GSH/GSSG, and MDA concentration were evaluated, respectively. The intracellular ROS generation in CCs was markedly elevated after CCs were treated with 20 and 50 μM Genipin (P < 0.05, Figure 2(a)), which might feedback induce an increase of Mn-SOD in 20 and 50 μM Genipin treated groups (P < 0.05, Figure 3(b)). And obviously decreases were detected in the ratio of reduced GSH to GSSG in 20 and 50 μM Genipin treated groups too (P < 0.05, Figure 2(d)). In contrast, the MDA concentration increased significantly after being treated with 50 μM Genipin as compared with control (P < 0.05, Figure 2(c)).


Effects of Mitochondrial Uncoupling Protein 2 Inhibition by Genipin in Human Cumulus Cells.

Ge H, Zhang F, Shan D, Chen H, Wang X, Ling C, Xi H, Huang J, Zhu C, Lv J - Biomed Res Int (2015)

Effects of UCP2 inhibited by Genipin on CCs apoptosis. (a) The caspase-3 protein was quantified by Western blotting. The proactive caspase-3 declined obviously, and the active caspase-3 markedly increased after UCP2 inhibition. (b) Caspase-3 gene was quantified by RT-PCR, which markedly increase after UCP2 inhibition. Con means control, asterisks indicate significant difference between control and treatment group at P < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556840&req=5

fig3: Effects of UCP2 inhibited by Genipin on CCs apoptosis. (a) The caspase-3 protein was quantified by Western blotting. The proactive caspase-3 declined obviously, and the active caspase-3 markedly increased after UCP2 inhibition. (b) Caspase-3 gene was quantified by RT-PCR, which markedly increase after UCP2 inhibition. Con means control, asterisks indicate significant difference between control and treatment group at P < 0.05.
Mentions: To determine effects of UCP2 on cellular redox status in CCs, Mn-SOD activity and the levels of the ROS, ratio of GSH/GSSG, and MDA concentration were evaluated, respectively. The intracellular ROS generation in CCs was markedly elevated after CCs were treated with 20 and 50 μM Genipin (P < 0.05, Figure 2(a)), which might feedback induce an increase of Mn-SOD in 20 and 50 μM Genipin treated groups (P < 0.05, Figure 3(b)). And obviously decreases were detected in the ratio of reduced GSH to GSSG in 20 and 50 μM Genipin treated groups too (P < 0.05, Figure 2(d)). In contrast, the MDA concentration increased significantly after being treated with 50 μM Genipin as compared with control (P < 0.05, Figure 2(c)).

Bottom Line: There were no significant differences in ATP and mitochondrial membrane potential levels in cumulus cells from UCP2 inhibiting groups as compared with the control.However, the ratio of reduced GSH to GSSG significantly declined after treatment with Genipin.But there was no significant difference in estradiol concentrations.

View Article: PubMed Central - PubMed

Affiliation: Reproductive Health Center, Second Affiliated Hospital of Wenzhou Medical University, 109 Xueyuan Road, Wenzhou, Zhejiang 325000, China.

ABSTRACT
UCP2 plays a physiological role by regulating mitochondrial biogenesis, maintaining energy balance, ROS elimination, and regulating cellular autophagy in numerous tissues. But the exact roles of UCP2 in cumulus cells are still not clear. Genipin, a special UCP2 inhibitor, was added into the cultural medium to explore the roles of UCP2 in human cumulus cells. There were no significant differences in ATP and mitochondrial membrane potential levels in cumulus cells from UCP2 inhibiting groups as compared with the control. The levels of ROS and Mn-SOD were markedly elevated after UCP2 inhibited Genipin. However, the ratio of reduced GSH to GSSG significantly declined after treatment with Genipin. UCP2 inhibition by Genipin also resulted in obvious increase in the active caspase-3, which accompanied the decline of caspase-3 mRNA. The level of progesterone in culture medium declined obviously after Genipin treatment. But there was no significant difference in estradiol concentrations. This study indicated that UCP2 is expressed in human cumulus cells and plays important roles on mediate ROS production, apoptotic process, and steroidogenesis, suggesting UCP2 may be involved in regulation of follicle development and oocyte maturation and quality.

No MeSH data available.