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 JBM  Vol.6 No.6 , June 2018
Effects of Endoplasmic Reticulum Stress on Pulmonary Hypertension in Rat Induced by Chronic Hypoxia and Hypercapnia
Abstract: Objective: To observe the pulmonary vascular remodeling in rats with pulmonary hypertension induced by hypoxia and hypercapnia, and to explore the role of endoplasmic reticulum stress in pulmonary hypertension. Methods: 1) 40 SD rats were randomly divided into four groups: normoxic control group (N), hypoxia hypercapnia group (HH), endoplasmic reticulum stress (ERS) inhibitor 4-phenyl butyric acid group (4-PBA), ERS pathway agonist tunicamycin group (TM). 2) The mean pulmonary arterial pressure (mPAP) and the right ventricular hypertrophy index (RV/(LV + S)) were measured in each group. 3) Identification of pulmonary artery smooth muscle cells (PASMCs) in each group by immunofluorescence α-SMA. 4) Morphological changes of lung tissue and pulmonary artery were observed by electron microscope. 5) The apoptotic index of PASMCs in each group was detected by TUNEL. 6) Reverse transcription polymerase chain reaction (RT-PCR) and Western Blot (WB) were used to detect the expression of ERS related protein and mRNA (GRP78, CHOP, JNK, Caspase-12) in each group. Results: 1) Compared with the N group, the mPAP, RV/(LV + S) and vascular wall area (WA)/total area (TA) value of HH group, 4-PBA group and TM group were increased (P < 0.01), and the vascular lumen area (LA)/TA values, PASMCs apoptosis index were significantly decreased. GRP78, CHOP, JNK, Caspase-12 expression were increased, and the differences were statistically significant. 2) Compared with the HH group, the mPAP, RV/(LV + S) and WA/TA of 4-PBA group were decreased (P < 0.01); the LA/TA value and PASMCs apoptosis index were increased (P < 0.05); and the mRNA and protein expression of CHOP, JNK, Caspase-12 and GRP78 had a significant decrease (P < 0.05). 3) Compared with the HH, the mPAP, RV/(LV + S) and WA/TA of TM group were increased (P < 0.05, P < 0.01), while LA/TA were decreased (P < 0.01); and  PASMCs apoptotic index was increased (P < 0.01). Meanwhile, the mRNA expression of Caspase-12, CHOP, JNK and GRP78 was increased to varying degrees (P < 0.05), and the protein expression of Caspase-12, CHOP and JNK was also increased significantly (P < 0.01). Conclusion: Hypoxia and hypercapnia induced pulmonary vascular remodeling may be related to the proliferation of PASMCs, and ERS related factors (JNK, Caspase12 and CHOP) are involved in the regulation of hypoxic hypercapnia.
Cite this paper: Zhang, C. , Zhang, J. , Wu, Y. , Dai, Y. , Ying, L. and Wang, W. (2018) Effects of Endoplasmic Reticulum Stress on Pulmonary Hypertension in Rat Induced by Chronic Hypoxia and Hypercapnia. Journal of Biosciences and Medicines, 6, 53-67. doi: 10.4236/jbm.2018.66004.
References

[1]   Farha, S., Hu, B., Comhair, S., et al. (2016) Mitochondrial Haplogroups and Risk of Pulmonary Arterial Hypertension. PLoS ONE, 11, e0156042.
https://doi.org/10.1371/journal.pone.0156042

[2]   O’Connell, C., Montani, D., Savale, L., et al. (2015) Chronic Thromboembolic Pulmonary Hypertension. Presse Medicale, 44, e409-e416.
https://doi.org/10.1016/j.lpm.2015.10.010

[3]   Hao, Y., Thakkar, V., Stevens, W., et al. (2015) A Comparison of the Predictive Accuracy of Three Screening Models for Pulmonary Arterial Hypertension in Systemic Sclerosis. Arthritis Research & Therapy, 17, 7.
https://doi.org/10.1186/s13075-015-0517-5

[4]   Baker, S.C., Shabir, S., Georgopoulos, N.T., et al. (2016) Ketamine-Induced Apoptosis in Normal Human Urothelial Cells: A Direct, N-Methyl-d-Aspartate Receptor-Independent Pathway Characterized by Mitochondrial Stress. American Journal of Pathology, 186, 1267-1277.
https://doi.org/10.1016/j.ajpath.2015.12.014

[5]   Touzet, O. and Philips, A. (2010) Resveratrol Protects against Protease Inhibitor-Induced Reactive Oxygen Species Production, Reticulum Stress and Lipid Raft Perturbation. AIDS, 24, 1437-1447.
https://doi.org/10.1097/QAD.0b013e32833a6114

[6]   Inan, S. and Wei, H. (2010) The Cytoprotective Effects of Dantrolene: A Ryanodine Receptor Antagonist. Anesthesia & Analgesia, 111, 1400-1410.
https://doi.org/10.1213/ANE.0b013e3181f7181c

[7]   Chida, A., Shintani, M., Matsushita, Y., et al. (2014) Mutations of NOTCH3 in Childhood Pulmonary Arterial Hypertension. Molecular Genetics & Genomic Medicine, 2, 229-239.
https://doi.org/10.1002/mgg3.58

[8]   Liu, J., Xiao, M., Li, J., et al. (2017) Activation of UPR Signaling Pathway Is Associated with the Malignant Progression and Poor Prognosis in Prostate Cancer. Prostate, 77, 274-281.
https://doi.org/10.1002/pros.23264

[9]   Poletto, V., Dragoni, S., Lim, D., et al. (2016) Endoplasmic Reticulum Ca(2+) Handling and Apoptotic Resistance in Tumor-Derived Endothelial Colony Forming Cells. Journal of Cellular Biochemistry, 117, 2260-2271.
https://doi.org/10.1002/jcb.25524

[10]   Xue, Q., Li, C., Chen, J., et al. (2016) The Protective Effect of the Endoplasmic Reticulum Stress-Related Factors BiP/GRP78 and CHOP/Gadd153 on Noise-Induced Hearing Loss in Guinea Pigs. Noise Health, 18, 247-255.
https://doi.org/10.4103/1463-1741.192481

[11]   Tardif, N., Salles, J., Guillet, C., et al. (2014) Muscle Ectopic Fat Deposition Contributes to Anabolic Resistance in Obese Sarcopenic Old Rats through eIF2alpha Activation. Aging Cell, 13, 1001-1011.
https://doi.org/10.1111/acel.12263

[12]   Kim, K., Kim, Y.H., Lee, S.H., et al. (2014) Effect of Exercise Intensity on Unfolded Protein Response in Skeletal Muscle of Rat. The Korean Journal of Physiology & Pharmacology, 18, 211-216.
https://doi.org/10.4196/kjpp.2014.18.3.211

[13]   Chen, H., Yang, H., Pan, L., et al. (2016) The Molecular Mechanisms of XBP-1 Gene Silencing on IRE1alpha-TRAF2-ASK1-JNK Pathways in Oral Squamous Cell Carcinoma under Endoplasmic Reticulum Stress. Biomedicine & Pharmacotherapy, 77, 108-113.
https://doi.org/10.1016/j.biopha.2015.12.010

[14]   Lakshmanan, A.P., Thandavarayan, R.A., Palaniyandi, S.S., et al. (2011) Modulation of AT-1R/CHOP-JNK-Caspase12 Pathway by Olmesartan Treatment Attenuates ER Stress-Induced Renal Apoptosis in Streptozotocin-Induced Diabetic Mice. European Journal of Pharmaceutical Sciences, 44, 627-634.
https://doi.org/10.1016/j.ejps.2011.10.009

[15]   Sari, F.R., Watanabe, K., Widyantoro, B., et al. (2010) Partial Inactivation of Cardiac 14-3-3 Protein in Vivo Elicits Endoplasmic Reticulum Stress (ERS) and Activates ERS-Initiated Apoptosis in ERS-Induced Mice. Cellular Physiology and Biochemistry, 26, 167-178.
https://doi.org/10.1159/000320548

[16]   Denzel, M.S., Storm, N.J., Gutschmidt, A., et al. (2014) Hexosamine Pathway Metabolites Enhance Protein Quality Control and Prolong Life. Cell, 156, 1167-1178.
https://doi.org/10.1016/j.cell.2014.01.061

[17]   De Juan-Sanz, J., Holt, G.T., Schreiter, E.R., et al. (2017) Axonal Endoplasmic Reticulum Ca2+ Content Controls Release Probability in CNS Nerve Terminals. Neuron, 93, 867-881.e6.
https://doi.org/10.1016/j.neuron.2017.01.010

[18]   To, M., Peterson, C.W., Roberts, M.A., et al. (2017) Lipid Disequilibrium Disrupts ER Proteostasis by Im-pairing ERAD Substrate Glycan Trimming and Dislocation. Molecular Biology of the Cell, 28, 270-284.
https://doi.org/10.1091/mbc.e16-07-0483

[19]   Han, J., Back, S.H., Hur, J., et al. (2013) ER-Stress-Induced Transcriptional Regulation Increases Protein Synthesis Leading to Cell Death. Nature Cell Biology, 15, 481-490.
https://doi.org/10.1038/ncb2738

[20]   Vega, H., Agellon, L.B. and Michalak, M. (2016) The Rise of Proteostasis Promoters. IUBMB Life, 68, 943-954.
https://doi.org/10.1002/iub.1576

[21]   Grall, S., Prunier-Mirebeau, D., Tamareille, S., et al. (2013) Endoplasmic Reticulum Stress Pathway Involvement in Local and Remote Myocardial Ischemic Conditioning. Shock, 39, 433-439.
https://doi.org/10.1097/SHK.0b013e31828e4f80

[22]   Hong, Y.P., Guo, W.Y., Wang, W.X., et al. (2016) 4-Phenylbutyric Acid Attenuates Pancreatic Beta-Cell Injury in Rats with Experimental Severe Acute Pancreatitis. International Journal of Endocrinology, 2016, Article ID: 4592346.

[23]   Conrad, N., Schwager, S.L., Carmona, A.K., et al. (2016) The Effect of Structural Motifs on the Ectodomain Shedding of Human Angiotensin-Converting Enzyme. Biochemical and Biophysical Research Communications, 481, 111-116.
https://doi.org/10.1016/j.bbrc.2016.10.155

[24]   Beretta, G.L., Benedetti, V., Cossa, G., et al. (2010) Increased Levels and Defective Glycosylation of MRPs in Ovarian Carcinoma Cells Resistant to Oxaliplatin. Biochemical Pharmacology, 79, 1108-1117.
https://doi.org/10.1016/j.bcp.2009.12.002

[25]   Liu, Z., Gu, H., Gan, L., et al. (2016) Reducing Smad3/ATF4 Was Essential for Sirt1 Inhibiting ER Stress-Induced Apoptosis in Mice Brown Adipose Tissue. Oncotarget, 8, 9267-9279.

[26]   Lopez-Sambrooks, C., Shrimal, S., Khodier, C., et al. (2016) Oli-gosaccharyltransferase Inhibition Induces Senescence in RTK-Driven Tumor Cells. Nature Chemical Biology, 12, 1023-1030.
https://doi.org/10.1038/nchembio.2194

[27]   Ling, Y.H., Li, T., Perez-Soler, R., et al. (2009) Activation of ER Stress and Inhibition of EGFR N-Glycosylation by Tunicamycin Enhances Susceptibility of Human Non-Small Cell Lung Cancer Cells to Erlotinib. Cancer Chemotherapy and Pharmacology, 64, 539-548.
https://doi.org/10.1007/s00280-008-0902-8

[28]   Han, X., Zhang, X., Li, H., et al. (2015) Tunicamycin Enhances the Antitumor Activity of Trastuzumab on Breast Cancer in Vitro and in Vivo. Oncotarget, 6, 38912-38925.
https://doi.org/10.18632/oncotarget.5334

[29]   Kim, H., Baek, C.H., Lee, R.B., et al. (2017) Anti-Fibrotic Effect of Losartan, an Angiotensin II Receptor Blocker, Is Mediated through Inhibition of ER Stress via Up-Regulation of SIRT1, Followed by Induction of HO-1 and Thioredoxin. International Journal of Molecular Sciences, 18, 305.
https://doi.org/10.3390/ijms18020305

[30]   Suyama, K., Watanabe, M., Sakabe, K., et al. (2011) Overexpression of GRP78 Protects Glial Cells from Endoplasmic Reticulum Stress. Neuroscience Letters, 504, 271-276.
https://doi.org/10.1016/j.neulet.2011.09.045

[31]   Pinto, B.A., Melo, T.M., Flister, K.F., et al. (2016) Early and Sustained Exposure to High-Sucrose Diet Triggers Hippocampal ER Stress in Young Rats. Metabolic Brain Disease, 31, 917-927.
https://doi.org/10.1007/s11011-016-9830-1

 
 
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