Berberine protects human renal proximal tubular cells from hypoxia/reoxygenation injury via inhibiting endoplasmic reticulum and mitochondrial stress pathways

Abstract

BackgroundIschemia/reperfusion injury plays a crucial role in renal transplantation, and represents a significant risk factor for acute renal failure and delayed graft function. The pathophysiological contribution of endoplasmic reticulum and mitochondria stress to ischemia/reperfusion injury has also been highlighted. Berberine (BBR) has been showed to attenuate ischemia/reperfusion injury by inhibiting oxidative stress. The study was carried out to investigate whether the pretreatment of BBR could reduce hypoxia/reoxygenation (H/R)-induced injury by inhibiting mitochondria stress and endoplasmic reticulum stress pathways.MethodsThe cultured human renal proximal tubular cell line HK-2 cells were exposed to 24 h hypoxia (5% CO2, 1% O2, 94% N2) followed by 3 h reoxygenation (5% CO2, 21% O2, 74% N2). And BBR was added to the culture medium 2h prior to the treatment. Then the cell viability, oxidative stress level, morphological change of apoptosis and apoptotic rate were determined. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax and cytochrome C involved in mitochondrial-dependent pathway and ER stress hallmarks such as glucose-regulated protein 78 and CCAAT/enhancer binding protein homologous protein.ResultsH/R produced dramatic injuries in HK-2 cells. The cell viability and the oxidative stress level in group H/R was significantly decreased. The classical morphological change of apoptosis was found, while the apoptotic rate and the expression of proteins involved in mitochondrial stress and endoplasmic reticulum stress pathways increased (p<0.05). Administration of BBR significantly inhibited these H/R induced changes (p<0.05).ConclusionThis study revealed that BBR pretreatment serves a protective role against H/R induced apoptosis of human renal proximal tubular cells, and the mechanism is related to suppression of mitochondrial stress and endoplasmic reticulum stress pathways.

Nephroprotective Role of Chrysophanol in Hypoxia/Reoxygenation-Induced Renal Cell Damage via Apoptosis, ER Stress, and Ferroptosis

“…As shown in Figure 2 A, H/R-induced ER stress was attenuated by chrysophanol treatment. Consistent with our findings, Huaier extract was found to downregulate the expression of CHOP and Bip in a thapsigargin-induced ER stress model of HK-2 cells [ 42 ]; the same result was also observed in other studies [ 27 ]. On the other hand, ferroptosis is well known to play a critical role in the pathology of AKI [ 19 ].…”

Nephroprotective Role of Chrysophanol in Hypoxia/Reoxygenation-Induced Renal Cell Damage via Apoptosis, ER Stress, and Ferroptosis

“…As shown in Figure 2 A, H/R-induced ER stress was attenuated by chrysophanol treatment. Consistent with our findings, Huaier extract was found to downregulate the expression of CHOP and Bip in a thapsigargin-induced ER stress model of HK-2 cells [ 42 ]; the same result was also observed in other studies [ 27 ]. On the other hand, ferroptosis is well known to play a critical role in the pathology of AKI [ 19 ].…”

A new combination of naringin and trimetazidine protect kidney Mitochondria dysfunction induced by renal Ischemia / Reperfusion injury in rat

“…These results were parallel to what was obtained for ROS generation with IR injury, showing the greatest damage. In vitro study in renal proximal cell demonstrated that hypoxia/reoxygenation injury caused mitochondrial stress and antioxidant treatment could reverse this change (Yu et al, 2013). The current study showed that renal IR was able to enhance ROS generation in kidney mitochondria.…”

Endoplasmic reticulum stress and its effects on renal tubular cells apoptosis in ischemic acute kidney injury

“…25 In parallel, two ER stress hallmarks, GRP78 and CHOP, showed increased protein levels after hypoxia and ER stress, and participated in hypoxia/reoxygenation induced apoptosis in human renal proximal tubular epithelial cell line (HK-2 cells). 49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis.…”

“…49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis. 48 There are other interventions being explored.…”

Nephroprotective Role of Chrysophanol in Hypoxia/Reoxygenation-Induced Renal Cell Damage via Apoptosis, ER Stress, and Ferroptosis

“…As shown in Figure 2 A, H/R-induced ER stress was attenuated by chrysophanol treatment. Consistent with our findings, Huaier extract was found to downregulate the expression of CHOP and Bip in a thapsigargin-induced ER stress model of HK-2 cells [ 42 ]; the same result was also observed in other studies [ 27 ]. On the other hand, ferroptosis is well known to play a critical role in the pathology of AKI [ 19 ].…”

A new combination of naringin and trimetazidine protect kidney Mitochondria dysfunction induced by renal Ischemia / Reperfusion injury in rat

“…These results were parallel to what was obtained for ROS generation with IR injury, showing the greatest damage. In vitro study in renal proximal cell demonstrated that hypoxia/reoxygenation injury caused mitochondrial stress and antioxidant treatment could reverse this change (Yu et al, 2013). The current study showed that renal IR was able to enhance ROS generation in kidney mitochondria.…”

Endoplasmic reticulum stress and its effects on renal tubular cells apoptosis in ischemic acute kidney injury

“…25 In parallel, two ER stress hallmarks, GRP78 and CHOP, showed increased protein levels after hypoxia and ER stress, and participated in hypoxia/reoxygenation induced apoptosis in human renal proximal tubular epithelial cell line (HK-2 cells). 49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis.…”

“…49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis. 48 There are other interventions being explored.…”

Nephroprotective Role of Chrysophanol in Hypoxia/Reoxygenation-Induced Renal Cell Damage via Apoptosis, ER Stress, and Ferroptosis

“…As shown in Figure 2 A, H/R-induced ER stress was attenuated by chrysophanol treatment. Consistent with our findings, Huaier extract was found to downregulate the expression of CHOP and Bip in a thapsigargin-induced ER stress model of HK-2 cells [ 42 ]; the same result was also observed in other studies [ 27 ]. On the other hand, ferroptosis is well known to play a critical role in the pathology of AKI [ 19 ].…”

A new combination of naringin and trimetazidine protect kidney Mitochondria dysfunction induced by renal Ischemia / Reperfusion injury in rat

“…These results were parallel to what was obtained for ROS generation with IR injury, showing the greatest damage. In vitro study in renal proximal cell demonstrated that hypoxia/reoxygenation injury caused mitochondrial stress and antioxidant treatment could reverse this change (Yu et al, 2013). The current study showed that renal IR was able to enhance ROS generation in kidney mitochondria.…”

Endoplasmic reticulum stress and its effects on renal tubular cells apoptosis in ischemic acute kidney injury

“…25 In parallel, two ER stress hallmarks, GRP78 and CHOP, showed increased protein levels after hypoxia and ER stress, and participated in hypoxia/reoxygenation induced apoptosis in human renal proximal tubular epithelial cell line (HK-2 cells). 49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis.…”

“…49 However, those parameters were significantly suppressed by berberine pretreatment. 49 It was also demonstrated that tauroursodeoxycholic acid (TUDCA) pretreatment had a nephroprotective effect on ischemia-induced AKI by inhibiting ER stress and by blocking GRP78 and CHOP expression, reducing Caspase 12 activation, and inhibiting cell apoptosis. 48 There are other interventions being explored.…”