The Mechanisms of the Herbal Components of CRSAS on HK‐2 Cells in a Hypoxia/Reoxygenation Model Based on Network Pharmacology

Ye, Naijing; Xie, Dengpiao; Yang, Bing; Li, Mingquan

doi:10.1155/2020/5352490

Cited by 6 publications

(5 citation statements)

References 27 publications

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“…Network pharmacology shows that the targets of P. igniarius in the treatment of HUA involve the regulation of the apoptotic process, ER stress and the ATF4 transcription factor. Furthermore, studies have shown that ROSinduced chronic ER stress produces a secondary increase in ROS, which usually leads to apoptosis of HK-2 cells [39][40][41]. ER stress is also closely associated with various renal cell injuries and is involved in mediating the development of renal diseases [42][43][44].…”

Section: Discussionmentioning

confidence: 99%

Network Pharmacology-Based Approach to Explore the Total Flavonoids of Phellinus igniarius against Hyperuricemia via Regulating the Nrf2/HO-1-ROS-ER Signaling Pathway

Lv,

Liu,

Tao

et al. 2023

Journal of Food Biochemistry

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Based on the antioxidant properties of Phellinus igniarius (DC. Ex Fr.) Quel (P. igniarius), this study aims to investigate the protective effect and mechanism of total flavonoids of Phellinus igniarius (TFPI) on the oxidative damage of HK-2 cells induced by monosodium urate (MSU). The GO and KEGG enrichment analyses predicted the potential targets and pathways of TFPI in the treatment of hyperuricaemia (HUA). We used MSU to stimulate HK-2 cells to establish a HUA model. Cell viability, lactate dehydrogenase (LDH) assay, and reactive oxygen species (ROS) assay were performed. Cell nuclear morphology was detected with DAPI and Annexin V-FITC. The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) was analyzed. The expression of Keap 1, Nrf2, HO-1, Bip, PERK, ATF4, CHOP, BAX, Bcl-2, and cleaved caspase 3 was performed with western blot. The results of network pharmacology showed that the potential targets and pathways were mainly associated with stimulus response, regulation of reactive oxygen species metabolic processes, response to oxidative stress, and regulation of the response to the endoplasmic reticulum (ER) stress pathway. The cell experiment proved that the survival rate of HK-2 cells was dramatically increased after treatment with TFPI. TFPI increased the activity of SOD and decreased the content of MDA and LDH, while scavenging ROS. TFPI increased the transfer of Nrf2 protein from the cytoplasm to the nucleus and decreased the expression level of Bip, PERK, ATF4, CHOP, BAX, and cleaved caspase 3 to attenuate apoptotic cells induced by MSU. TFPI has a good protective effect on MSU-induced oxidative damage in HK-2 cells and can reduce cell apoptosis by regulating ROS-mediated ER stress.

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Section: Discussionmentioning

confidence: 99%

Network Pharmacology-Based Approach to Explore the Total Flavonoids of Phellinus igniarius against Hyperuricemia via Regulating the Nrf2/HO-1-ROS-ER Signaling Pathway

Lv,

Liu,

Tao

et al. 2023

Journal of Food Biochemistry

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“…The appropriate IC50 concentration of TGP was determined according to the results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The hypoxia/reoxygenation (H/R) model in vitro was established by the following steps: 21 HK-2 cells were cultured in the medium free of glucose and serum under the conditions of 1% O 2 , 5% CO 2 and 94% N 2 for 24 h. Thereafter, the cells were cultured in the DMEM containing 10% FBS under the conditions of 21% O 2 , 5% CO 2 and 74% N 2 for 12 h.…”

Section: Methodsmentioning

confidence: 99%

Total Glucosides of Paeony Inhibited Autophagy and Improved Acute Kidney Injury Induced by Ischemia-Reperfusion via the lncRNA TUG1/miR-29a/PTEN Axis

Chang¹,

Zhang²,

Xu³

et al. 2021

DDDT

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Objective: Total glucosides of paeony (TGP) has been proven to affect anti-inflammatory, immunomodulatory and hypoxia tolerance. This study investigates the effect of TGP on autophagy in acute kidney injury (AKI) induced by ischemia-reperfusion (I/R). Methods: Rat model of AKI induced by I/R was established. Rats were administered with TGP at different doses by oral gavage. The contents of BUN, creatinine, NGAL, Kim-1 and IL-18 were detected. The levels of inflammatory factors (TNF-α, IL-1β and IL-6) and autophagy were measured. The expressions of lncRNA TUG1, miR-29a and PTEN were detected and their binding relationships were verified. I/R rat model with overexpressed TUG1 was established to explore the effect of TGP on kidney injury and autophagy. The hypoxia/reoxygenation (HR) model of HK-2 cells and the HR model of HK-2 cells overexpressing TUG1 and low-expressing PTEN were established. Results: TGP decreased the contents of BUN, creatinine, NGAL, Kim-1 and IL-18, and reduced the levels of inflammatory factors. LncRNA TUG1 and PTEN were downregulated, and miR-29a was upregulated in kidney tissues. The binding relationships between lncRNA TUG1 and miR-29a, and miR-29a and PTEN were confirmed. TGP suppressed PTEN expression via the lncRNA TUG1/miR-29a axis. Overexpressing lncRNA TUG1 attenuated the protective effect of TGP on AKI and autophagy in HK-2 cells. TGP improved cell viability and inhibited the autophagy in HR model of HK-2 cells via lncRNA TUG1/miR-29a/PTEN axis. Conclusion: TGP inhibited autophagy and improved AKI induced by I/R via the lncRNA TUG1/miR-29a/PTEN axis.

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“…e renal tubular epithelial HK-2 cell line was purchased from Shanghai Aiyan Biotechnology Co., Ltd. (Shanghai, China). e HK-2 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum; then, they were placed in incubators at 37°C with 5% CO 2 and passaged 1 time every 3-5 days [15]. After being synchronized with culture for 24 h, the HK-2 cells were placed in 24-well plates at a density of 2 × 10 4 -5 × 10 4 cells per well, and the medium then was covered with tape and sealed for 5 h to simulate the process of ischemia-reperfusion injury.…”

Section: Establishment Model Of H/r and Groupingmentioning

confidence: 99%

“…ere are few reports on the mechanism of SE for the treatment of AKI. To the best of our knowledge, only one study has reported that the SE components of rhubarb, Salvia miltiorrhiza, Astragalus membranaceus, and Safflower (CRSAS) can reduce HK-2 cell apoptosis by inhibiting the expression of CHOP [15].…”

Section: Introductionmentioning

confidence: 99%

Shenshuaikang Enema, a Chinese Herbal Remedy, Inhibited Hypoxia and Reoxygenation‐Induced Apoptosis in Renal Tubular Epithelial Cells by Inhibiting Oxidative Damage‐Dependent JNK/Caspase‐3 Signaling Pathways Using Network Pharmacology

Lü

Luo

et al. 2020

Evidence-Based Complementary and Alternative Medicine

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Background. Acute kidney injury (AKI) is a common clinically critical illness with serious consequences for the patients. Shenshuaikang enema (SE) is a Chinese herbal compound that is used to treat AKI in clinical practice. However, its mechanism of action remains unclear. Aim. The aim of this study was to investigate the therapeutic effect of SE and explore the molecular mechanisms using network pharmacology and in vitro experiments. Materials and Methods. The herb-component-target network was constructed based on network pharmacology. The predicted targets and pathways were validated using in vitro experiments. A renal tubular epithelial cell line (HK-2 cells) was exposed to hypoxia and reoxygenation (H/R) using air-tight conditions for five hours and treated with different concentrations of SE (25%, 50%, and 75%) to assess cell viability and apoptosis and determine the optimal experimental dose. Subsequently, H/R-injured HK-2 cells were pretreated with the optimal SE dose and then randomly divided into three groups, the SE, SE-SP600125 (inhibitor of JNK), and SE-NAC (antioxidant) groups. The cell vitality, apoptosis, and death were evaluated using the cell counting kit 8 (CCK8) and carboxyfluorescein succinimidyl ester/propidium iodide (CFSF/PI) staining. The apoptosis-related protein JNK and Caspase-3 were assessed by Western blot. Expression of JNK and Caspase-3 genes was analyzed using real-time quantitative polymerase chain reaction (RT-qPCR). Results. 123 active components and 226 targets were identified from four herbs that composed the herb-compound-target network based on transcriptomics and network pharmacology analyses. The KEGG pathway analyses revealed that the mitochondrial apoptosis pathway was involved in the therapeutic AKI effects of SE. Cell vitality of H/R-induced HK-2 cells was obviously increased when treating them with SE, and the apoptosis was significantly inhibited, especially in the SE (50%) group at 4 and 12 h after modeling. Pretreatment with antioxidant NAC obviously prevented cell death compared to the SE (50%) group, while no obvious reduction of apoptosis was observed in the SP600125 group. JNK expression level was significantly increased in the SE (50%) group compared to the SP600125 ( P < 0.01 ) and the NAC group ( P < 0.05 ). Caspase-3 was downregulated in the SE (50%) group compared to the SP600125 ( P < 0.01 ) and NAC group ( P < 0.05 ). Caspase-3 activation in the SP600125 group was higher than that in the NAC group ( P < 0.05 ). Moreover, the oxidative damage-dependent JNK/Caspase-3 pathway was identified in the H/R-injured HK-2 cells by inhibiting the JNK activation and oxidative damage. Conclusions. Our findings suggested that the H/R-triggered apoptosis in HK-2 cells was abrogated by SE by upregulating the oxidative damage-dependent JNK to trigger suppression of Caspase-3.

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The Mechanisms of the Herbal Components of CRSAS on HK‐2 Cells in a Hypoxia/Reoxygenation Model Based on Network Pharmacology

Cited by 6 publications

References 27 publications

Network Pharmacology-Based Approach to Explore the Total Flavonoids of Phellinus igniarius against Hyperuricemia via Regulating the Nrf2/HO-1-ROS-ER Signaling Pathway

Network Pharmacology-Based Approach to Explore the Total Flavonoids of Phellinus igniarius against Hyperuricemia via Regulating the Nrf2/HO-1-ROS-ER Signaling Pathway

Total Glucosides of Paeony Inhibited Autophagy and Improved Acute Kidney Injury Induced by Ischemia-Reperfusion via the lncRNA TUG1/miR-29a/PTEN Axis

Shenshuaikang Enema, a Chinese Herbal Remedy, Inhibited Hypoxia and Reoxygenation‐Induced Apoptosis in Renal Tubular Epithelial Cells by Inhibiting Oxidative Damage‐Dependent JNK/Caspase‐3 Signaling Pathways Using Network Pharmacology

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