Cardioprotective effect of recombinant adeno-associated virus mediated nerve growth factor gene transfection in diabetic rats
(2.Dept of Anesthesiology, the Second Hospital of Shanxi Medical University, Taiyuan, China 030001)
【Abstract】 Aim To investigate the feasibility of recombinant adeno-associated virus serotype9 (rAAV9) mediated nerve growth factor (NGF) transfection in diabetic rats by intramyocardial injection, and to confirm the protective effect of this method on diabetic cardiac autonomic neuropathy. Methods At the 4th week after the establishment of diabetic model, the recombinant adeno-associated virus (rAAV9-NGF) carrying NGF gene was injected into the heart of rats. Five weeks later, the distributions of NGF, calcitonin gene-related peptide (calcitonin gene-related peptide, CGRP) and nerve fibers were detected by immunofluorescence. Hemodynamic parameters were used to evaluate cardiac function, and ELISA method was used to detect the expression of NGF, CGRP in myocardium. Results The exogenous gene NGF mediated by rAAV9 could be stably transfected and over-expressed in the myocardium of diabetic rats. The left ventricular systolic pressure, heart rate, and maximal rate of increase of left ventricular pressure significantly decreased, the left ventricular end-diastolic pressure and the maximal decrease rate of left ventricular pressure significantly increased, and the cardiac function was improved. The NGF, CGRP proteins in myocardial tissues were up-regulated, and the immunofluorescence showed that the decrease of NGF, CGRP and nerve fibers in myocardial tissues could be partially reversed after transfection. Conclusions Intramyocardial injection of NGF gene can effectively enhance the expression of NGF and promote the growth of nerve fiber, exerting a protective effect in diabetic rat heart.
【Keywords】 nerve growth factor; calcitonin gene-related peptide; transfection; recombinant adeno-associated virus; diabetes peripheral neuropathy; heart disease;
 Fisher V L, Tahrani A A. Cardiac autonomic neuropathy in patients with diabetes mellitus: current perspectives [J]. Diabetes Metab Syndr Obes, 2017, 10: 419–34.
 Marlin M C, Li G. Biogenesis and function of the NGF/TrkA signaling endosome [J]. Int Rev Cell Mol Biol, 2015, 314 (6): 239–57.
 Ieda M, Kanazawa H, Ieda Y, et al. Nerve growth factor is critical for cardiac sensory innervation and rescues neuropathy in diabetic hearts [J]. Circulation, 2006, 114 (22): 2351–63.
 Sheng Y, Zhu L. The crosstalk between autonomic nervous system and blood vessels [J]. Int J Physiol Pathophysiol Pharmacol, 2018, 10 (1): 17.
 Que W Z, Chen J M. Gateway technical supported construction of a recombinant adenovirus vector pAd-NK4 [J]. Chin Pharmacol Bull, 2011, 27 (4): 462–7 (in Chinese).
 Pacak C A, Mah C S, Thattaliyath B D, et al. Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo[J]. Circ Res, 2006, 99 (4): 3–9.
 Li L, Liu Z Z.Progress in the study of nerve growth factor and diabetic peripheral neuropathy [J]. Section Endocrinol Foreign Med Sci, 2004, 24 (z1): 30–3 (in Chinese).
 Chen Y F, Zhang L K, Wang R L, et al. Role of adenovirus-mediated cardiac-specific RIP140 overexpression in cardiac function and inflammation pathway [J]. Chin Pharmacol Bull, 2017, 33 (8): 1068–72 (in Chinese).
 Oakley I, Emond L. Diabetic cardiac autonomic neuropathy and anesthetic management: review of the literature [J]. AANA J, 2011, 79 (6): 473–9.
 Landowski L M, Dyck P J, Engelstad J, et al. Axonopathy in peripheral neuropathies: mechanisms and therapeutic approaches for regeneration [J]. J Chem Neuroanat, 2016, 76 (Pt A): 19–27.
 Lu D, Liu Y X, Jing L, et al. Curative effect evaluation and pharmacological analysis of mouse nerve growth factor for diabetic peripheral neuropathy [J]. Chin J Mod Drug Appl, 2016, 10 (9): 16–7 (in Chinese).
 Hirose M, Kuroda Y, Murata E. NGF/TrkA signaling as a therapeutic target for pain [J]. Pain Pract, 2016, 16 (2): 175–82.
 Yue W. The expression of CGRP and NGF at stage of acute myocardial infarction [D]. Taiyuan: Shanxi Medical University, 2011 (in Chinese).