Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation.
<h4>Aims</h4>Reactive oxygen species (ROS) can induce renal damage when renal ischemia-reperfusion occurs as a result of factors such as renal transplant and shock. It is well established that oxidant damage plays an important role in the mechanism of ischemia-reperfusion. Resveratrol (R...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2025-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0327135 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849340410821869568 |
|---|---|
| author | Shicheng Luo Yuning Chen Xuyi Ma Feng Tian Xiaozhou He |
| author_facet | Shicheng Luo Yuning Chen Xuyi Ma Feng Tian Xiaozhou He |
| author_sort | Shicheng Luo |
| collection | DOAJ |
| description | <h4>Aims</h4>Reactive oxygen species (ROS) can induce renal damage when renal ischemia-reperfusion occurs as a result of factors such as renal transplant and shock. It is well established that oxidant damage plays an important role in the mechanism of ischemia-reperfusion. Resveratrol (RSV) is known for its antioxidant properties. However, in our investigations we found that RSV could aggravate human proximal tubular cells (HK-2 cells) damage when exposed to H2O2. Consequently, our findings suggested that RSV may potentially aggravate renal damage in the context of renal ischemia-reperfusion.<h4>Methods</h4>CCK-8 reagent kits were utilized to assess cell proliferation. Western blot analysis was employed to determine the expression level of protein about γ-H2ax, PTEN, AKT, P-AKT and Cleave-caspase3. Additionally, the process of apoptosis was examined using flow cytometry.<h4>Result</h4>The CCK-8 assays results revealed a dose-dependent inhibition of cell proliferation when HK-2 cells were exposed to varying concentrations of H2O2. Furthermore, when HK-2 cells were cultured with 250uM H2O2 and different concentrations of RSV, the CCK-8 results indicated a significant reduction in HK-2 cell viability with increasing RSV dosage. Flow cytometry analysis of HK-2 cells exposed to 250uM H2O2 and 10uM RSV demonstrated that RSV exacerbated apoptosis in H2O2-induced HK-2 cells. Western blot analysis revealed that RSV inhibited the expression of P-AKT and exacerbated damage to H2O2-induced HK-2 cells. Subsequently, we selected 10uM MK for further investigation, with CCK-8 results showed a significant reduction in HK-2 cell viability when the AKT phosphorylation process was inhibited by MK. Western blot analysis indicated that the cleave-caspase-3 was further activated when the AKT phosphorylation process was inhibited by MK. Flow cytometry demonstrated a significant increase in the apoptosis of HK-2 cells when the AKT phosphorylation process was inhibited by MK. In the case of P-AKT overexpression, Western blot analysis showed a significant increase in P-AKT expression, while a significant decrease in cleave-caspase-3. Flow cytometry revealed a significant decrease in the apoptosis of HK-2 cells. Finally, Western blot analysis demonstrated a decrease in P-AKT levels along with the expression of PTEN.<h4>Conclusions</h4>Our research substantiated that RSV could aggravate H2O2 induced HK-2 cells damage by upregulating the expression of PTEN and inhibiting AKT phosphorylation. It suggested that RSV may deteriorate renal damage in the context of renal ischemia-reperfusion injury. |
| format | Article |
| id | doaj-art-b7abfa5e6d224f2a93f6d7dc6eb0226d |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-b7abfa5e6d224f2a93f6d7dc6eb0226d2025-08-20T03:43:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01207e032713510.1371/journal.pone.0327135Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation.Shicheng LuoYuning ChenXuyi MaFeng TianXiaozhou He<h4>Aims</h4>Reactive oxygen species (ROS) can induce renal damage when renal ischemia-reperfusion occurs as a result of factors such as renal transplant and shock. It is well established that oxidant damage plays an important role in the mechanism of ischemia-reperfusion. Resveratrol (RSV) is known for its antioxidant properties. However, in our investigations we found that RSV could aggravate human proximal tubular cells (HK-2 cells) damage when exposed to H2O2. Consequently, our findings suggested that RSV may potentially aggravate renal damage in the context of renal ischemia-reperfusion.<h4>Methods</h4>CCK-8 reagent kits were utilized to assess cell proliferation. Western blot analysis was employed to determine the expression level of protein about γ-H2ax, PTEN, AKT, P-AKT and Cleave-caspase3. Additionally, the process of apoptosis was examined using flow cytometry.<h4>Result</h4>The CCK-8 assays results revealed a dose-dependent inhibition of cell proliferation when HK-2 cells were exposed to varying concentrations of H2O2. Furthermore, when HK-2 cells were cultured with 250uM H2O2 and different concentrations of RSV, the CCK-8 results indicated a significant reduction in HK-2 cell viability with increasing RSV dosage. Flow cytometry analysis of HK-2 cells exposed to 250uM H2O2 and 10uM RSV demonstrated that RSV exacerbated apoptosis in H2O2-induced HK-2 cells. Western blot analysis revealed that RSV inhibited the expression of P-AKT and exacerbated damage to H2O2-induced HK-2 cells. Subsequently, we selected 10uM MK for further investigation, with CCK-8 results showed a significant reduction in HK-2 cell viability when the AKT phosphorylation process was inhibited by MK. Western blot analysis indicated that the cleave-caspase-3 was further activated when the AKT phosphorylation process was inhibited by MK. Flow cytometry demonstrated a significant increase in the apoptosis of HK-2 cells when the AKT phosphorylation process was inhibited by MK. In the case of P-AKT overexpression, Western blot analysis showed a significant increase in P-AKT expression, while a significant decrease in cleave-caspase-3. Flow cytometry revealed a significant decrease in the apoptosis of HK-2 cells. Finally, Western blot analysis demonstrated a decrease in P-AKT levels along with the expression of PTEN.<h4>Conclusions</h4>Our research substantiated that RSV could aggravate H2O2 induced HK-2 cells damage by upregulating the expression of PTEN and inhibiting AKT phosphorylation. It suggested that RSV may deteriorate renal damage in the context of renal ischemia-reperfusion injury.https://doi.org/10.1371/journal.pone.0327135 |
| spellingShingle | Shicheng Luo Yuning Chen Xuyi Ma Feng Tian Xiaozhou He Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. PLoS ONE |
| title | Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. |
| title_full | Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. |
| title_fullStr | Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. |
| title_full_unstemmed | Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. |
| title_short | Resveratrol aggravated H2O2-induced the HK-2 cell damage by inhibiting AKT phosphorylation. |
| title_sort | resveratrol aggravated h2o2 induced the hk 2 cell damage by inhibiting akt phosphorylation |
| url | https://doi.org/10.1371/journal.pone.0327135 |
| work_keys_str_mv | AT shichengluo resveratrolaggravatedh2o2inducedthehk2celldamagebyinhibitingaktphosphorylation AT yuningchen resveratrolaggravatedh2o2inducedthehk2celldamagebyinhibitingaktphosphorylation AT xuyima resveratrolaggravatedh2o2inducedthehk2celldamagebyinhibitingaktphosphorylation AT fengtian resveratrolaggravatedh2o2inducedthehk2celldamagebyinhibitingaktphosphorylation AT xiaozhouhe resveratrolaggravatedh2o2inducedthehk2celldamagebyinhibitingaktphosphorylation |