投稿系统
Direct evidence of VEGF-mediated neuroregulation and afferent explanation of blood pressure dysregulation during angiogenic therapy
doi: 10.2478/fzm-2021-0015
-
Abstract:
Objective Oncocardiology is increasingly hot research field/topic in the clinical management of cancer with anti-angiogenic therapy of vascular endothelial growth factor (VEGF) that may cause cardiovascular toxicity, such as hypertension via vascular dysfunction and attenuation of eNOS/NO signaling in the baroreflex afferent pathway. The aim of the current study was to evaluate the potential roles of VEGF/VEGF receptors (VEGFRs) expressed in the baroreflex afferent pathway in autonomic control of blood pressure (BP) regulation. Methods The distribution and expression of VEGF/VEGFRs were detected in the nodose ganglia (NG) and nucleus of tractus solitary (NTS) using immunostaining and molecular approaches. The direct role of VEGF was tested by NG microinjection under physiological and hypertensive conditions. Results Immunostaining data showed that either VEGF or VEGFR2/VEGFR3 was clearly detected in the NG and NTS of adult male rats. Microinjection of VEGF directly into the NG reduced the mean blood pressure (MBP) dose-dependently, which was less dramatic in renovascular hypertension (RVH) rats, suggesting the VEGF-mediated depressor response by direct activation of the 1st-order baroreceptor neurons in the NG under both normal and disease conditions. Notably, this reduced depressor response in RVH rats was directly caused by the downregulation of VEGFR2, which compensated the up regulation of VEGF/VEGFR3 in the NG during the development of hypertension. Conclusion It demonstrated for the first time that the BP-lowering property of VEGF/VEGFRs signaling via the activation of baroreflex afferent function may be a common target/pathway leading to BP dysregulation in anti-angiogenic therapy. -
Figure 1. Immunohistochemical characterization of the expression and distribution of VEGF, VEGFR2 and VEGFR3 in the NG and NTS.
(A) The results of conventional immunohistochemical experiments carried out with specific antibodies against DAPI (blue), HCN1 (red), and VEGF, VEGFR2 or VEGFR3 (green) to detect the cellular/subcellular distributions of VEGF, VEGFR2 and VEGFR3 in the NG of adult male SD rats. Yellow and white arrowheads represent HCN1-positive (myelinated A-type afferents) and HCN1-negative (unmyelinated C-type afferents) neurons, respectively. Scale bar = 50 μm. (B) The representative immunostaining images of whole brainstem section showing the distribution of VEGF, VEGFR2 and VEGFR3 in the NTS. Sol: solitary tract; the scale bar = 2.0 mm. SolL: nucleus of the solitary tract (Sol); SolVL: -ventrolateral part; SolV: -ventral part; SolM: -medial part; SolDM: -dorsomedial part; SolIM: -intermediate part. The fluorescent bar represents the expression level. The negative staining as a control for both NG and NTS is presented in supplemental materials.
Figure 2. VEGF-mediated BP reduction in control and RVH rats following direct microinjection of VEGF into the NG.
(A and B) Changes in BP and body weight of Ctrl and RVH rats during 4 weeks after surgery; (C) Representative recordings of VEGF-medicated BP reduction following administration of a series of doses of VEGF via direct microinjection into the left side of the NG in control (Ctrl) and RVH model rats. Dotted lines represent the time points of microinjections; (D) Summarized data of VEGF-mediated BP reduction with different doses of VEGF; (E) Summarized data showing the recovery time of VEGF-mediated BP reduction. The data are expressed as mean ± SD and n = 5 rats for each group, *P < 0.05 and **P < 0.01 vs. Ctrl.
Figure 3. Changes in cardiac function, blood pressure, and baroreflex sensitivity in RVH rats.
(A) Cardiac parameters determined by echocardiograph in control and RVH rats. Data are presented as mean ± SD, n = 4, *P < 0.05 vs. control (Ctrl). PSLAX: parasternal long axis; LVIDs/LVIDd: systolic/diastolic left ventricular internal diameter; IVSs/IVSd: systolic/diastolic left ventricular septum thickness; LVPWs/LVPWd: systolic/diastolic left ventricular posterior wall; EF: ejection fraction; FS: fractional shortening. (B) Representative recordings of mean arterial blood pressure (MAP, purple) and heart rate (blue) collected from Ctrl (n = 5) and RVH (n = 5) rats treated with 1, 3, and 10 μg/kg of SNP and PE, respectively. Scale bars are applied to all recordings. (C-D) The summarized changes of BRS (ΔHR/ΔMABP, bpm/mmHg) after treatment with SNP or PE at different concentrations. Data are expressed as means ± SD. *P < 0.05 and **P < 0.01 vs. Ctrl.
Figure 4. Relative mRNA and protein expression levels of VEGF, VEGFR2 and VEGFR3 in the NG and NTS.
(A-C) Relative mRNA and protein expression levels (fold change) and representative protein bends of VEGF, VEGFR2, and VEGFR3 in the NG of control and RVH model rats compared with internal control (GAPDH); (D-F) Relative mRNA and protein expression levels (fold change) and representative protein bends of VEGF, VEGFR2, and VEGFR3 in the NTS of control and RVH model rats compared with internal control (GAPDH). Data are presented as mean ± SD; *P < 0.05 and **P < 0.01 vs. Ctrl, n = 4 duplicated tests and tissue specimens collected from 6 rats for both PCR and Western blot analyses.
Figure 5. The expression alterations of VEGF, VEGFR2 and VEGFR3 in the kidney.
(A) Expression alterations (WB and PCR) of VEGF in the kidney of control and RVH rats compared with the internal control (GAPDH). Data are presented as mean ± SD; *P < 0.05 vs. control, n = 4; (B) Expression changes (WB and PCR) of VEGFR2 in the kidney of control and RVH rats compared with the internal control (GAPDH). Data are presented as mean ± SD; *P < 0.05 vs. control, n = 4; (C) Expression alterations (WB and PCR) of VEGFR3 in the kidney of control and RVH rats compared with the internal control (GAPDH). Data are presented as mean ± SD; *P < 0.05 vs. control, n = 4.
-
[1] Itatani Y, Yamamoto T, Zhong C, et al. Suppressing neutrophil-dependent angiogenesis abrogates resistance to anti-VEGF antibody in a genetic model of colorectal cancer. Proc Natl Acad Sci USA, 2020; 117(35): 21598-21608. doi: 10.1073/pnas.2008112117 [2] Chong W Q, Lim C M, Sinha A K, et al. Integration of antiangiogenic therapy with cisplatin and gemcitabine chemotherapy in patients with nasopharyngeal carcinoma. Clin Cancer Res, 2020; 26(20): 5320-5328. doi: 10.1158/1078-0432.CCR-20-1727 [3] Stitzlein L, Rao P, Dudley R. Emerging oral VEGF inhibitors for the treatment of renal cell carcinoma. Expert Opin Investig Drugs, 2019; 28(2): 121-130. doi: 10.1080/13543784.2019.1559296 [4] Versmissen J, Mirabito Colafella K M, Koolen S L W, et al. Vascular cardio-oncology: vascular endothelial growth factor inhibitors and hypertension. Cardiovasc Res, 2019; 115(5): 904-914. doi: 10.1093/cvr/cvz022 [5] Touyz R M, Lang N N, Herrmann J, et al. Recent advances in hypertension and cardiovascular toxicities with vascular endothelial growth factor inhibition. Hypertension, 2017; 70(2): 220-226. doi: 10.1161/HYPERTENSIONAHA.117.08856 [6] Jiang L, Ping L, Yan H, et al. Cardiovascular toxicity induced by anti-VEGF/VEGFR agents: a special focus on definitions, diagnoses, mechanisms and management. Expert Opin Drug Metab Toxicol, 2020; 16(4): 823-835. doi: 10.1080/17425255.2020.1787986?scroll=top [7] Balci S, Sahin O, Ozcaliskan S, et al. Immediate changes in blood pressure during intravitreal anti-VEGF agents' applications in exudative age-related macular degeneration patients. Int Ophthalmol, 2020; 40(10): 2515-2522. doi: 10.1007/s10792-020-01431-3 [8] Dobbin S J H, Cameron A C, Petrie M C, et al. Toxicity of cancer therapy: what the cardiologist needs to know about angiogenesis inhibitors. Heart, 2018; 104(24): 1995-2002. doi: 10.1136/heartjnl-2018-313726 [9] Yang R, Ogasawara A K, Zioncheck T F, et al. Exaggerated hypotensive effect of vascular endothelial growth factor in spontaneously hypertensive rats. Hypertension, 2002; 39(3): 815-820. doi: 10.1161/hy0302.105398 [10] Hariawala M D, Horowitz J R, Esakof D, et al. VEGF improves myocardial blood flow but produces EDRF-mediated hypotension in porcine hearts. J Surg Res, 1996; 63(1): 77-82. doi: 10.1006/jsre.1996.0226 [11] Horowitz J R, Rivard A, Van der Zee R, et al. Vascular endothelial growth factor/vascular permeability factor produces nitric oxide-dependent hypotension. Evidence for a maintenance role in quiescent adult endothelium. Arterioscler Thromb Vasc Biol, 1997; 17(11): 2793-2800. doi: 10.1161/01.ATV.17.11.2793 [12] Murohara T, Horowitz J R, Silver M, et al. Vascular endothelial growth factor/vascular permeability factor enhances vascular permeability via nitric oxide and prostacyclin. Circulation, 1998; 97(1): 99-107. doi: 10.1161/01.CIR.97.1.99 [13] Dong Z C, Wu M M, Zhang Y L, et al. The vascular endothelial growth factor trap aflibercept induces vascular dysfunction and hypertension via attenuation of eNOS/NO signaling in mice. Acta Pharmacol Sin, 2020; Dec 10. doi10.1038/s41401-020-00596-1. Online ahead of print. [14] Santa Cruz Chavez G C, Li B Y, Glazebrook P A, et al. An afferent explanation for sexual dimorphism in the aortic baroreflex of rat. Am J Physiol Heart Circ Physiol, 2014; 307: H910-H921. doi: 10.1152/ajpheart.00332.2014 [15] Liu Y, Zhou J Y, Zhou Y H, et al. Unique expression of angiotensin type-2 receptor in sex-specific distribution of myelinated Ah-type baroreceptor neuron contributing to sex-dimorphic neurocontrol of circulation. Hypertension, 2016; 67: 783-791. doi: 10.1161/HYPERTENSIONAHA.115.06815 [16] Wen X, Yu X, Huo R, et al. Serotonin-mediated cardiac analgesia via Ah-type baroreceptor activation contributes to silent angina and asymptomatic infarction. Neuroscience, 2019; 411: 150-163. doi: 10.1016/j.neuroscience.2019.05.045 [17] Chen P, Xu B, Feng Y, et al. FGF-21 ameliorates essential hypertension of SHR via baroreflex afferent function. Brain Res Bull, 2020; 154: 9-20. doi: 10.1016/j.brainresbull.2019.10.003 [18] Feng Y, Liu Y, Cao P X, et al. Estrogen-dependent microRNA-504 expression and related baroreflex afferent neuroexcitation via negative regulation on KCNMB4 and KCa1.1 beta4-subunit expression. Neuroscience, 2020; 442: 168-182. doi: 10.1016/j.neuroscience.2020.07.003 [19] Li Y, Feng Y, Liu L, et al. The baroreflex afferent pathway plays a critical role in H2S-mediated autonomic control of blood pressure regulation under physiological and hypertensive conditions. Acta Pharmacol Sin, 2020; 42(6): 898-908. http://www.nature.com/articles/s41401-020-00549-5?utm_content=null&utm_medium=cpc&utm_source=trendmd [20] Liu Y, Zhao S Y, Feng Y, et al. Contribution of baroreflex afferent pathway to NPY-mediated regulation of blood pressure in rats. Neurosci Bull, 2020; 36(4): 396-406. doi: 10.1007/s12264-019-00438-w [21] Wang X, Li G, Liu L, et al. Effects of extreme temperatures on cause-specific cardiovascular mortality in China. Int J Envir Res Public Health, 2015; 12(12): 16136-16156. doi: 10.3390/ijerph121215042 [22] Yang L, Li L, Lewington S, et al. Outdoor temperature, blood pressure, and cardiovascular disease mortality among 23 000 individuals with diagnosed cardiovascular diseases from China. Eur Heart J, 2015; 36(19): 1178-1185. doi: 10.1093/eurheartj/ehv023 [23] Sharma A, Sharma T, Panwar M S, et al. Colder environments are associated with a greater cancer incidence in the female population of the United States. Tumour Biol, 2017; 39(10): 1010428317724784. http://europepmc.org/abstract/MED/29022494 [24] Gomez-Acebo I, Llorca J, Dierssen T. Cold-related mortality due to cardiovascular diseases, respiratory diseases and cancer: a case-crossover study. Public Health, 2013; 127(3): 252-258. doi: 10.1016/j.puhe.2012.12.014 [25] Guan J, Zhao M, He C, et al. Anti-hypertensive action of fenofibrate via UCP2 upregulation mediated by PPAR activation in baroreflex afferent pathway. Neurosci Bull, 2019; 35(1): 15-24. doi: 10.1007/s12264-018-0271-1 [26] Zeng W Z, Marshall K L, Min S, et al. PIEZOs mediate neuronal sensing of blood pressure and the baroreceptor reflex. Science, 2018; 362(6413): 464-467. doi: 10.1126/science.aau6324 [27] Li B Y, Schild J H. Electrophysiological and pharmacological validation of vagal afferent fiber type of neurons enzymatically isolated from rat nodose ganglia. J Neurosci Methods, 2007; 164(1): 75-85. doi: 10.1016/j.jneumeth.2007.04.003 [28] Li B Y, Schild J H. Patch clamp electrophysiology in nodose ganglia of adult rat. J Neurosci Methods, 2002; 115(2): 157-167. doi: 10.1016/S0165-0270(02)00010-9 [29] Andresen M C, Kunze D L. Nucleus tractus solitarius--gateway to neural circulatory control. Ann Rev Physiol, 1994; 56(1): 93-116. doi: 10.1146/annurev.ph.56.030194.000521 [30] Zhang Y Y, Yan Z Y, Qu M Y, et al. KCa1.1 is potential marker for distinguishing Ah-type baroreceptor neurons in NTS and contributes to sex-specific presynaptic neurotransmission in baroreflex afferent pathway. Neurosci Lett, 2015; 604: 1-6. doi: 10.1016/j.neulet.2015.07.023 [31] Xu W X, Yu J L, Feng Y, et al. Spontaneous activities in baroreflex afferent pathway contribute dominant role in parasympathetic neurocontrol of blood pressure regulation. CNS Neurosci Ther, 2018; 24: 1219-1230. doi: 10.1111/cns.13039 -
下载:
点击查看大图
计量
- 文章访问数: 64
- HTML全文浏览量: 35
- PDF下载量: 7
- 被引次数: 0
