Apelin-13 Alleviates Hexavalent Chromium-Induced Myocardial Injury by Inhibiting ROS-mediated Oxidative Damage and Regulating Multiple Signals

doi:10.21203/rs.3.rs-2080260/v2

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Apelin-13 Alleviates Hexavalent Chromium-Induced Myocardial Injury by Inhibiting ROS-mediated Oxidative Damage and Regulating Multiple Signals

https://doi.org/10.21203/rs.3.rs-2080260/v2

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Hexavalent chromium (Cr (VI)) from environmental pollution shows severe toxicity towards human organs by causing oxidative damage. Apelin-13 may inhibit oxidative damage by blocking ROS accumulation and regulating multiple signals. However, whether apelin-13 can attenuate Cr (VI)-induced toxicity has not been explored. Herein, the protective effect and molecular mechanism of apelin-13 against K₂Cr₂O₇-induced cardiotoxicity in vitro and in vivo were evaluated. Results show that apelin-13 significantly inhibited K₂Cr₂O₇-induced H9c2 cytotoxicity and apoptosis, followed by the attenuated PARP cleavage and caspase activation. Further investigation revealed that apelin-13 co-treatment effectively suppressed K₂Cr₂O₇-induced oxidative damage by inhibiting ROS accumulation. Moreover, apelin-13 co-treatment dramatically normalized MAPKs and PI3K/AKT pathways in K₂Cr₂O₇-treated H9c2 cells. Importantly, apelin-13 administration in vivo effectively attenuated myocardial fibrosis, improved angiogenesis, and inhibited myocardial abnormal proliferation and apoptosis. Ultimately, it alleviated K₂Cr₂O₇-induced myocardial injury in rats. Taken together, our findings validated the strategy of the use of apelin-13 to effectively combat Cr (VI)-induced myocardial injury.

Hexavalent chromium

apelin-13

oxidative damage

myocardial injury

apoptosis

Heavy metal pollution is rapidly increasing in the environment, and its negative effects can largely be attributed to hexavalent chromium (Cr (VI)) [1]. Exposure to Cr (VI) can trigger oxidative damage, DNA damage, cell apoptosis, and inflammation [2]. Nevertheless, the mechanisms by which the metal pollutant of Cr (VI) mediate cardiovascular disease pathogenesis have not been completely understood.

K₂Cr₂O₇, as an extremely complicated factor, can lead to irreversible pathological change, which greatly induce the morbidity and mortality of many cardiovascular diseases [3]. MAPK and PI3K/Akt signaling pathways are closely related to cell proliferation, differentiation, survival, and apoptosis, especially under various pathological states, such as chronic Cr (VI) exposure-induced heart injury [4]. Therefore, apelin-13 may enhance the cardio-protective effect of heart under K₂Cr₂O₇-induced pathological states, which may be regulated by the MAPK and PI3K/Akt signaling pathways.

Apelin-13, as a ligand, is widely involved in regulating the cardiovascular process because of its strong biological activity of peptide hormone, and it can promote the inner walls of blood vessels and tube formation of microvascular endothelial cells and regulate the blood pressure and myocardial injury [5]. However, the protective effects and the underlying mechanism of apelin-13 against Cr (VI)-induced heart injury remains unclear. Hence, in the present study, we investigate the direct effects of apelin-13 on the K₂Cr₂O₇-induced cardiotoxicity in vitro and in vivo.

1. Materials and agents

H9c2 embryonic rat cardiac cells were obtained from the Chinese Academy of Sciences (Shanghai, China). Apelin-13 was obtained from GL Biochem Shanghai (Shanghai, China). K₂Cr₂O₇ was obtained from Tianjin Tianli Chemical Reagent (Tianjin, China). The following primary antibodies were obtained from Abcam: cleaved-PARP (1:1,000), active-Caspase-7 (1:1,000), Active-Caspase-3 (1:500), Ser15-p53 (1:1,000), Total-p53 (1:800), Ser139-H2A (1:5,000), Thr183-JNK(1:500), Total-JNK (1:1,000), Thr180-p38 (1:1,000), Total-p38 (1:1,000), Thr202-ERK (1:1,000), Total-ERK (1:1,000), Ser473-AKT (1:5,000), Total-AKT (1:500), eNOS (1:50), and β-actin (1:1,000). Active-Caspase-9 (1:500) was obtained from BioVision. Ser428-ATR (1:1,000) was obtained from CST. ROS (1:100), and superoxide anion (1:80) was obtained from Sigma.

2. Cell Culture And Drug Treatment

H9c2 cells were cultured in DMEM-F12 containing 20% FBS in an incubator 5% CO₂ at 37°C. To attain 70–80% confluence, we treated the cells with K₂Cr₂O₇ for 48 h. Some cultures were pre-treated for 1 h with apelin-13. The control cells were incubated without apelin-13 and K₂Cr₂O₇.

3. Cell Viability Assay

Cell viability assay was performed via Cell Counter Kit-8 (CCK-8) assay.

4. Detection Of Apoptosis

Treated cells were harvested, fixed, and permeabilized with 70% ethanol, washed with PBS, and incubated with 0.5 mL of 50 µg/mL PI solution containing 20 U/mL RNase A. Flow cytometry analysis was carried out using a FACS caliber.

5. Western Blot Analysis

Cells were homogenized in RIPA lysis buffer containing protease inhibitor PMSF. Total protein was measured using BCA, size separated by SDS-PAGE, and electrotransferred onto nitrocellulose membrane. The nitrocellulose blots were blocked with 5% non-fat dry milk in TBST buffer, and then incubated with the desired primary antibodies. After washing thrice with TBST buffer for 5 min, the nitrocellulose membranes were incubated with goat anti-rabbit IgG or goat anti-mouse IgG second antibody for 2 h. After washing thrice with TBST buffer for 5 min, the resulting immunocomplex bands were visualized using chemiluminescence photographic detection with Immobilon Western Chemiluminescent HRP Substrate.

6. Immunofluorescence Staining

Plated cells were fixed with pre-cooled methanol/acetone (v/v, 1:1) at room temperature for 5 min, and then rinsed in PBS. Fixed cells were blocked in 10% goat serum with 0.1% Triton X-100 for 30 min, and then incubated with primary antibodies overnight at 4°C. After washing thrice with PBS, cells were incubated with fluorescein-labeled (FITC) or rhodamine-labeled (TRITC) antibody for 1 h at room temperature. Cell nuclei were counterstained with 4′,6-diamidino-2-phenylindole. Samples were observed under a fluorescence microscope with a high-resolution digital camera.

7. In vivo study

The study was conducted on adult male Sprague Dawley (SD) rats with body weight of 180–220 g. The rats were contained in standard vivarium conditions and fed on a standard diet. Animal use in the experiment was performed according to the guidelines established by the Guide for the Care and Use of Laboratory Animals of The Affiliated Taian City Central Hospital of Qingdao University. The procedure was approved by the Local Animal Ethics Committee. Rats were maintained under conventional conditions (room temperature, 22.5–23.0°C; relative humidity, 50–70%; 12 h day/night cycle) with free access to food and water.

The animals were randomly divided into four groups (10 animals/group), namely, the control group (normal saline), K₂Cr₂O₇ group (8 mg/kg), K₂Cr₂O₇ (8 mg/kg) + apelin-13 (4 mg/kg) group, and apelin-13 (4 mg/kg) group. K₂Cr₂O₇ was dissolved in normal saline and administered via the caudal vein. Model group animals were exposed to K₂Cr₂O₇ for 2 weeks. Protective group animals were co-treated with K₂Cr₂O₇ (8 mg/kg) + apelin-13 (4 mg/kg) for 2 weeks. The control group received an equal volume of normal saline.

8. Histopathological Evaluation

The tissue specimens were fixed with 10% neutral formalin and processed according to the standard histological methods. In total, 5–7-micron paraffin-embedded sections were stained with hematoxylin-eosin and picrofuchsin. The specimens were studied under a light microscope AxioLab A1 at 200× magnification.

In all experimental groups, a blinded semi-quantitative four-point scale from 0 to 3 (0 ¼ none, 1 ¼ weak, 2 ¼ moderate, and 3 ¼ strong) was assigned to calculate the severity of K₂Cr₂O₇-induced heart tissue in terms of the fibrotic area, cardiomyocyte hypertrophy area, eNOS, inflammatory cells, active-caspase-3, microvessel density, and Ki67.

9. Statistical Analyses

For multiple comparisons, statistical analysis was performed using one-way ANOVA, followed by Tukey’s multiple comparison tests.

1. Apelin-13 inhibited Cr (VI)-induced cytotoxicity of H9c2 cells

K₂Cr₂O₇-induced cell death was observed in the H9c2 cardiomyoblast cell line. The concentration of K₂Cr₂O₇ was set to 8 µM for all experiments, and nearly 50% of the K₂Cr₂O₇-treated cells died within 48 h (Fig. 1A). Cells were treated with various concentrations of apelin-13 in the group treated with 8 µM K₂Cr₂O₇. Apelin-13 exhibited a protective effect in a dose-dependent manner on the cell viability against the damage caused by K₂Cr₂O₇ (Fig. 1B). Results show that treatment with apelin-13 for 1 h markedly reversed the morphological damage caused by K₂Cr₂O₇ compared with that of the control group, and this result is consistent with the findings of CCK-8 assay (Fig. 1C). These results indicate that apelin-13 can block K₂Cr₂O₇-induced H9c2 cell death and morphological damage.

2. Apelin-13 Attenuated Cr (Vi)-induced Oxidative Damage And Apoptosis In H9c2 Cells

Results indicate DNA damage and abnormal apoptosis of H9c2 cells related to K₂Cr₂O₇, which are potential factors that induced cardiotoxicity. As shown in Figs. 2A and 2B, K₂Cr₂O₇ treatment alone induced an increase in DNA damage and apoptosis cytokines levels compared with the control group in a dose-dependent manner, whereas treatment with apelin-13 decreased the Cleaved-PARP, Active-Caspase-7, Active-Caspase-9, and Active-Caspase-3 levels (Fig. 2C).

3. Apelin-13 Blocked Cr (Vi)-induced Ros Stimulation

To further explore the mechanism of K₂Cr₂O₇-induced toxicity of H9c2 cells, we examined ROS generation and superoxide anion levels. In the K₂Cr₂O₇-induced toxicity in H9c2 cells, the cells displayed an increase in ROS and superoxide anion levels in a time-dependent manner (Fig. 3A). K₂Cr₂O₇ also remarkably increased in ATR, p53, and H2A in a time-dependent manner compared with the control group (Fig. 3B). However, Western blot analysis results confirmed that apelin-13 treatment blocked ATR, p53, and H2A (Fig. 3C). These results revealed that apelin-13 may suppress ROS-mediated dysfunction of K₂Cr₂O₇-treated H9c2 cells.

4. Apelin-13 Normalized The Mapk And Pi3k/akt Pathway

MAPKs and PI3K/AKT play important role in the proliferation, survival, and apoptosis of H9c2, which contribute to K₂Cr₂O₇-induced cardiotoxicity. As shown in Figs. 4A and 4B, K₂Cr₂O₇ treatment significantly stimulated the expression of p-JNK, p-p38, and p-ERK and downregulated the expression of p-Akt in a time-dependent manner. However, apelin-13 significantly decreased the phosphorylation of JNK, p38, and ERK and increased the phosphorylation of p-Akt in H9c2 cells (Figs. 4C and 4D). Results suggest that apelin-13 may attenuate K₂Cr₂O₇-induced cardiotoxicity by inhibiting ROS-mediated DNA damage and regulating MAPK and PI3K/AKT pathways. Thus, apelin-13 maybe a novel way for the treatment of K₂Cr₂O₇-mediated cardiotoxicity.

5. Apelin-13 ameliorates Cr (VI)-induced-induced heart injury in vivo

To determine whether apelin-13 is involved in K₂Cr₂O₇-induced cardiotoxicity in vivo, we used Masson staining, WGA staining, eNOS immunohistochemical, and H&E staining to explore the role of apelin-13 in the reversal of K₂Cr₂O₇-induced toxicity in heart tissues and the possible mechanism of action. Results show that apelin-13 remarkably inhibited the K₂Cr₂O₇-induced fibrotic area of heart tissue by Masson staining (Fig. 5A). Subsequently, apelin-13 effectively attenuated K₂Cr₂O₇-induced cardiotoxicity by inhibiting cardiomyocyte hypertrophy area by WGA staining (Fig. 5B). Further investigation revealed that apelin-13 dramatically suppressed K₂Cr₂O₇-induced inflammatory response by increasing eNOS (Fig. 5C). H&E staining was used to evaluate the cardiac inflammatory cell level, and apelin-13 substantially inhibited K₂Cr₂O₇-induced inflammatory cells by H&E staining, thus confirming apelin-13 anti-inflammation (Fig. 5D). These results suggest that apelin-13 may prevent K₂Cr₂O₇-induced cardiotoxicity.

6. Apelin-13 Attenuates Kcro-induced Heart Tissue Death

In the present study, we examined whether apelin-13 could reverse K₂Cr₂O₇-induced cardiotoxicity in vivo, studied the changes in active-caspase-3 positive cells, and determined CD34 and Ki67 positive cells. The results indicated that apelin-13 effectively attenuated K₂Cr₂O₇-induced dysfunction of active-caspase-3 (Fig. 6A). Moreover, apelin-13 treatment displayed improved K₂Cr₂O₇-induced microvessel density (Fig. 6B). In addition, apelin-13 substantially attenuated K₂Cr₂O₇-induced Ki67 (Fig. 6C). These results indicate that apelin-13 may inhibit K₂Cr₂O₇-induced heart tissue toxicity.

7. Apelin-13 Showed No Side Effects On The Main Organs Of Rats

To determine whether apelin-13 itself promotes heart, liver, spleen, lung, kidney, and brain in the absence of other stimuli, we assessed the growth of these tissues in response to various physiologically relevant concentrations of apelin-13 (2, 4, and 8mg/kg) by H&E staining in SD rats. Results show that apelin-13 treatment showed no toxicity towards heart, liver, spleen, lung, kidney, and brain after 48 h (Fig. 7).

In the present study, K₂Cr₂O₇-induced toxicity in cardiovascular disease was used to further investigate the mechanisms for the cardio-protective effects of apelin-13 in H9c2 cardiomyocyte cell cultures and adult male SD rats. Results show that apelin-13 may attenuate K₂Cr₂O₇-induced toxicity in cardiac tissue by inhibiting ROS-mediated DNA damage and regulating ROS initiated multiple signaling pathways, such as MAPKs and PI3K/AKT.

K₂Cr₂O₇-induced toxicity in H9c2 cells is a common cell model of Cr (VI) exposure to evaluate the cardio-protective profiles of bioactive reagents [6]. Apelin-13 can regulate the cardiovascular system [7]. Hence, the potential of apelin-13 was tested for its myocardiaprotective effect against toxicity induced by the toxins K₂Cr₂O₇ in H9c2 cells. We assessed K₂Cr₂O₇-induced cardiotoxicity through CCK-8 and confirmed that K₂Cr₂O₇ significantly decreased cell viability. However, treatment with different concentrations of apelin-13 markedly attenuated the decrease in cell viability. H9c2 displays cell shrinkage and nuclear condensation under K₂Cr₂O₇ stimuli, and these apoptotic features were detected by phase contract imaging. Interestingly, in the present study, apelin-13 treatment completely inhibited these apoptotic features. The results indicate that apelin-13 may inhibit K₂Cr₂O₇-induced apoptosis.

Oxidative stress is as an essential incentive of K₂Cr₂O₇-induced cardiac injury [8]. DNA damage activates Cleaved-PARP and induces and accelerates the apoptosis of myocardial cells [9]. In response to the caspase stimulus, caspase-9, caspase-3, and caspase-7 have distinct roles during intrinsic myocardial cells apoptosis [10]. “Initiator” caspase-9 is required for ROS production and can activate “effector” caspase-3 and caspase-7, while “effector” caspase-3, as the dominant executioner caspase, results in efficient apoptosis execution, and “effector” caspase-7 is responsible for cell detachment by cleaving various actin and cytoskeleton components during intrinsic myocardial cell death [11]. Thus, cleaved-PARP, caspase-9, caspase-3, and caspase-7 form a feedback loop to regulate upstream and downstream functions [12]. In these situations, apoptotic stimulation results in DNA damage [13]. Results show that apelin-13 treatment dramatically reversed K₂Cr₂O₇-induced DNA damage by inhibiting the expression levels of Cleaved-PARP and caspase activation.

DNA damage and myocardial cell apoptosis stimulate ROS overproduction [14]. Free radicals can cause tissue damage and worsen the Cr (VI) exposure of myocardial cells [15]. ROS induced by K₂Cr₂O₇ impairs Cleaved-PARP and caspases, resulting in the collapse of superoxide anion, thereby leading to ATR, which constitutes a significant factor for the activation of p53; subsequently, H₂A is activated, thus inducing DNA damage to inhibit cell viability and leading to myocardial cell apoptosis [16]. According to the results of the present study, apelin-13 ameliorated the deleterious effects of ROS by inhibiting the expression levels of ATR, p53, and H₂A in H9c2 cells treated with K₂Cr₂O₇.

Further investigation indicated that apelin-13 remarkably suppressed K₂Cr₂O₇-induced cardiotoxicity and blocked or activated the downstream signaling pathways. ROS overproduction activates the MAPK signaling pathway, and both pathways modulate cardiac pathophysiological signal transduction [17]. An increasing number of studies has proposed that p38 MAPK as a key target involve in cell biological response to an external stimulus, such as the activation of the p53 apoptotic pathway [18]. The JNK MAPK pathway contributes to functional activation of ATR in response to oxidative stress, resulting in the lack of p53 [19]. Similar to p38 and JNK MAPK, ERK MAPK plays a pivotal role in various cellular responses to ROS overproduction and DNA damage and regulates the activity of H₂A to conduct cell apoptosis [20]. The PI3K/Akt signaling pathway participates in vascular homeostasis and angiogenesis, can block the fibrotic area, and protect cardiomyocyte area; subsequently, eNOS phosphorylation occurs, leading to the regulation of myocardial inflammatory responses [21]. The PI3K/Akt signaling pathway plays an important role in the Cr (VI) exposure of heart, in which the elimination of p-Akt protein promotes cell death. The PI3K/Akt pathway plays a crucial role in the regulation of energy homeostasis, cell growth, and apoptosis, especially in cardioprotection [22]. In the present study, K₂Cr₂O₇ treatment causes the phosphorylation of JNK, p38, and ERK but inhibits the phosphorylation of AKT. Apelin-13 treatment effectively could K₂Cr₂O₇-induced dysfunction of MAPKs and PI3K/Akt signaling pathways. The results suggest that apelin-13 prevents K₂Cr₂O₇-induced toxicity via the inactivation of the MAPK and the activation of the PI3K/Akt pathways.

Further studies are required to evaluate the functional effect of apelin-13 against K₂Cr₂O₇ in vivo and to further test its cardio-protective potential. SD rats were used to explore the protective role of apelin-13 against the cardiotoxicity of K₂Cr₂O₇ exposure and reveal the potential molecular mechanism. In the present study, apelin-13 inhibited inflammation, further ameliorating heart damage, and ultimately protecting the heart from Cr (VI) poisoning.

Apoptotic heart tissue can form fibrotic area and destroy the cardiomyocyte area, which further promote heart tissue inflammatory response. Our studies have confirmed that apelin-13 treatment obviously attenuated K₂Cr₂O₇-induced fibrotic area and cardiomyocyte hypertrophy area.

Cardiac inflammation is important in the pathogenesis of Cr (VI) exposure [23]. eNOS plays a major role in the toxicity of heart tissue [24]. The results demonstrated that apelin-13 treatment counteracted the K₂Cr₂O₇-induced decrease in eNOS expression. Treatment with apelin-13 could substantially inhibit inflammatory cells. Our data show that apelin-13 alleviated active caspase-3 and Ki67 and increased microvessel density induced by K₂Cr₂O₇. These results confirm the protection of apelin-13 against K₂Cr₂O₇ exposure and provide a new perspective to understand the mechanism of apelin-13 to protect against heart injury.

Subsequently, the effect of apelin-13 toward the heart, liver, spleen, lung, kidney, and brain in the absence of other stimuli were examined. Importantly, apelin-13 alone do not cause these tissues injury.

In conclusion, results show that apelin-13 exerts its cardioprotective effect on Cr (VI)-induced death in cardiac tissue through direct scavenging of intracellular ROS induced by K₂Cr₂O₇ and inhibiting ROS-mediated DNA damage and downstream signaling pathways MAPK activation and PI3K/Akt inhibition (Fig. 8). Our findings validated the strategy of the use of apelin-13 is a potential novel pharmaceutical target for Cr (VI)-induced cardiovascular disease.

Acknowledgments

This work was supported by the Natural Science Foundation of Shandong Province under Grant number ZR2021MH089.

Author Contribution Statement

Pu Zhang, Rong Fu, and Hengshu Liu are the first authors as well as wrote the main manuscript text and contributed equally to this study. Jianfeng Xue, Luhua Yin, and Chao Cui are the correspond authors and guided the experiment. Wang Li, Yingying Zhang, and Na Song prepared figures 1-6. Hui Liu and Na Yao prepared figures 7-8. All authors reviewed the manuscript.

Competing Interest Statement

The authors have declared no competing interest.

Data Availability Statement

All data included in this study are available upon reasonable request by contact with the corresponding author.

Fu Y, Liu Y, Liu Y, Wang Y, Zhu M, Lin W, Li M, Liu Y, He M, Yu L, Wang J. Relationship between cumulative exposure to metal mixtures and heart rate among Chinese preschoolers. Chemosphere. 2022 Aug; 300: 134548.
Morvaridzadeh M, Estêvão MD, Qorbani M, Heydari H, Hosseini AS, Fazelian S, Belančić A, Persad E, Rezamand G, Heshmati J. The effect of chromium intake on oxidative stress parameters: A systematic review and meta-analysis. J Trace Elem Med Biol. 2022 Jan; 69: 126879.
Yang D, Yang Q, Fu N, Li S, Han B, Liu Y, Tang Y, Guo X, Lv Z, Zhang Z. Hexavalent Chromium Causes Apoptosis and Autophagy by Inducing Mitochondrial Dysfunction and Oxidative Stress in Broiler Cardiomyocytes. Chemosphere. 2021 Feb; 264(Pt 2): 128547.
Zhang P, Wang AP, Yang HP, Ai L, Zhang HJ, Wang YM, Bi YL, Fan HH, Gao J, Zhang HY, Liu JZ. Apelin-13 attenuates high glucose-induced calcifification of MOVAS cells by regulating MAPKs and PI3K/AKT pathways and ROS-mediated signals. Biomed Pharmacother. 2020 Aug; 128: 110271.
Yang M, Song JJ, Yang XC, Zhong GZ, Zhong JC. MiRNA-122-5p inhibitor abolishes angiotensin II-mediated loss of autophagy and promotion of apoptosis in rat cardiofibroblasts by modulation of the apelin-AMPK-mTOR signaling. In Vitro Cell Dev Biol Anim. 2022 Feb; 58(2): 136-148.
Li J, Zheng X, Ma X, Xu X, Du Y, Lv Q, Li X, Wu Y, Sun H, Yu L, Zhang Z. Melatonin protects against chromium(VI)-induced cardiac injury via activating the AMPK/Nrf2 pathway. J Inorg Biochem. 2019 Aug; 197: 110698.
Hu J, Huo S, Dou S, Jiang W, Li Y. Protective effect of Apelin/APJ system on lipopolysaccharide-related cardiac dysfunction. Gen Physiol Biophys. 2021 May; 40(3): 161-171.
Awoyomi OV, Adeoye YD, Oyagbemi AA, Ajibade TO, Asenuga ER, Gbadamosi IT, Ogunpolu BS, Falayi OO, Hassan FO, Omobowale TO, Arojojoye OA, Ola-Davies OE, Saba AB, Adedapo AA, Oguntibeju OO, Yakubu MA. Luteolin mitigates potassium dichromate-induced nephrotoxicity, cardiotoxicity and genotoxicity through modulation of Kim-1/Nrf2 signaling pathways. Environ Toxicol. 2021 Nov; 36(11): 2146-2160.
Ma M, Ma X, Cui J, Guo Y, Tang X, Chen C, Zhu Y, Cui C, Wang G. Cyclosporin A Protected Cardiomyocytes Against Oxidative Stress Injury by Inhibition of NF-κB Signaling Pathway. Cardiovasc Eng Technol. 2019 Jun;10(2):329-343.
Zeng C, Jiang W, Yang X, He C, Wang W, Xing J. Pretreatment with Total Flavonoid Extract from Dracocephalum Moldavica L. Attenuates Ischemia Reperfusion-induced Apoptosis. Sci Rep. 2018 Nov 30; 8(1): 17491.
Van Ngo H, Robertin S, Brokatzky D, Bielecka MK, Lobato-Márquez D, Torraca V, Mostowy S. Septins promote caspase activity and coordinate mitochondrial apoptosis. Cytoskeleton (Hoboken). 2022 Apr 23.
Song P, Huang H, Ma Y, Wu C, Yang X, Choi HY. Davidone C Induces the Death of Hepatocellular Carcinoma Cells by Promoting Apoptosis and Autophagy. Molecules. 2021 Aug 28; 26(17): 5219.
Alves R, Gonçalves AC, Jorge J, Almeida AM, Sarmento-Ribeiro AB. Combination of Elacridar with Imatinib Modulates Resistance Associated with Drug Efflux Transporters in Chronic Myeloid Leukemia. Biomedicines. 2022 May 17; 10(5): 1158.
Xiong L, Huang J, Wang S, Yuan Q, Yang D, Zheng Z, Wu Y, Wu C, Gao Y, Zou L, Hu G. Yttrium chloride-induced cytotoxicity and DNA damage response via ROS generation and inhibition of Nrf2/PPARγ pathways in H9c2 cardiomyocytes. Arch Toxicol. 2022 Mar; 96(3): 767-781.
Li H, Shi J, Gao H, Yang X, Fu Y, Peng Y, Xia Y, Zhou D. Hexavalent Chromium Causes Apoptosis and Autophagy by Inducing Mitochondrial Dysfunction and Oxidative Stress in Broiler Cardiomyocytes. Biol Trace Elem Res. 2022 Jun; 200(6): 2866-2875.
Men H, Cai H, Cheng Q, Zhou W, Wang X, Huang S, Zheng Y, Cai L. The regulatory roles of p53 in cardiovascular health and disease. Cell Mol Life Sci. 2021 Mar; 78(5): 2001-2018.
Wu HH, Meng TT, Chen JM, Meng FL, Wang SY, Liu RH, Chen JN, Ning B, Li Y, Su GH. Asenapine maleate inhibits angiotensin II-induced proliferation and activation of cardiac fibroblasts via the ROS/TGFβ1/MAPK signaling pathway. Biochem Biophys Res Commun. 2021 May 14; 553: 172-179.
Liu F, Fan LM, Geng L, Li JM. p47 phox-Dependent Oxidant Signalling through ASK1, MKK3/6 and MAPKs in Angiotensin II-Induced Cardiac Hypertrophy and Apoptosis. Antioxidants (Basel). 2021 Aug 26; 10(9): 1363.
Shi T, van Soest DMK, Polderman PE, Burgering BMT, Dansen TB. DNA damage and oxidant stress activate p53 through differential upstream signaling pathways. Free Radic Biol Med. 2021 Aug 20; 172:298-311.
Long Q, Xiao X, Yi P, Liu Y, Varier KM, Rao Q, Song J, Qiu J, Wang C, Liu W, Gajendran B, He Z, Liu S, Li Y. L20, a Calothrixin B analog, induces intrinsic apoptosis on HEL cells through ROS/γ-H2AX/p38 MAPK pathway. Biomed Pharmacother. 2021 May; 137: 111336.
Ning W, Li S, Yang W, Yang B, Xin C, Ping X, Huang C, Gu Y, Guo L. Blocking exosomal miRNA-153-3p derived from bone marrow mesenchymal stem cells ameliorates hypoxia-induced myocardial and microvascular damage by targeting the ANGPT1-mediated VEGF/PI3k/Akt/eNOS pathway. Cell Signal. 2021 Jan; 77:109812.
Xing JJ, Mi XJ, Hou JG, Cai EB, Zheng SW, Wang SH, Wang Z, Chen C, Li W. Maltol mitigates cisplatin-evoked cardiotoxicity via inhibiting the PI3K/Akt signaling pathway in rodents in vivo and in vitro. Phytother Res. 2022 Apr; 36(4): 1724-1735.
Al-Rasheed NM, Attia HA, Mohamed RA, Al-Rasheed NM, Al-Amin MA. Preventive effects of selenium yeast, chromium picolinate, zinc sulfate and their combination on oxidative stress, inflammation, impaired angiogenesis and atherogenesis in myocardial infarction in rats. J Pharm Pharm Sci. 2013;16(5):848-67.
Musicante M, Kim HH, Chen Y, Liao F, Bhattacharya SK, Lu L, Sun Y.Regulation of endothelial nitric oxide synthase in cardiac remodeling. Int J Cardiol. 2022 May 30:S0167-5273(22)00664-7.

No competing interests reported.

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Apelin-13 Alleviates Hexavalent Chromium-Induced Myocardial Injury by Inhibiting ROS-mediated Oxidative Damage and Regulating Multiple Signals

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Abstract

Figures

Introduction

Material And Methods

1. Materials and agents

2. Cell Culture And Drug Treatment

3. Cell Viability Assay

4. Detection Of Apoptosis

5. Western Blot Analysis

6. Immunofluorescence Staining

7. In vivo study

8. Histopathological Evaluation

9. Statistical Analyses

Results

1. Apelin-13 inhibited Cr (VI)-induced cytotoxicity of H9c2 cells

2. Apelin-13 Attenuated Cr (Vi)-induced Oxidative Damage And Apoptosis In H9c2 Cells

3. Apelin-13 Blocked Cr (Vi)-induced Ros Stimulation

4. Apelin-13 Normalized The Mapk And Pi3k/akt Pathway

5. Apelin-13 ameliorates Cr (VI)-induced-induced heart injury in vivo

6. Apelin-13 Attenuates Kcro-induced Heart Tissue Death

7. Apelin-13 Showed No Side Effects On The Main Organs Of Rats

Discussion

Declarations

References

Additional Declarations

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