垂体腺苷酸环化酶激活肽38对急性放射性心肌损伤的防护作用

doi:10.3969/j.issn.1674-8115.2021.02.001

上海交通大学学报(医学版) ›› 2021, Vol. 41 ›› Issue (2): 129-133.doi: 10.3969/j.issn.1674-8115.2021.02.001

垂体腺苷酸环化酶激活肽38对急性放射性心肌损伤的防护作用

李欢¹(), 易培强¹, 苏筠¹, 陈培战², 许赬¹, 曹璐¹, 陈佳艺¹, 李敏¹()

^1.上海交通大学医学院附属瑞金医院肿瘤放射治疗科，上海 200025
^2.上海交通大学医学院附属瑞金医院北院中心实验室，上海 201801

收稿日期:2020-06-01 出版日期:2021-02-28 发布日期:2021-02-28
通讯作者: 李敏 E-mail:lihuan3579@163.com;lm11866@rjh.com.cn
作者简介:李欢（1991—），女，住院医师，博士；电子信箱：lihuan3579@163.com。
基金资助:
国家重点研发计划(2016YFC0105409);国家自然科学基金面上项目(81602791);上海市科学技术委员会科技创新行动计划(19411950900);上海市教育委员会高峰高原学科建设计划(20171904)

Protective effect of pituitary adenylate cyclase-activating polypeptide 38 on acute radiation-induced myocardial injury

Huan LI¹(), Pei-qiang YI¹, Jun SU¹, Pei-zhan CHEN², Cheng XU¹, Lu CAO¹, Jia-yi CHEN¹, Min LI¹()

^1.Department of Radiation Oncology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
^2.Clinical Research Center, Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine, Shanghai 201801, China

Received:2020-06-01 Online:2021-02-28 Published:2021-02-28
Contact: Min LI E-mail:lihuan3579@163.com;lm11866@rjh.com.cn
Supported by:
National Key Research and Development Program of China(2016YFC0105409);National Natural Science Foundation of China(81602791);Scientific and Technological Innovation Action Plan of Science and Technology Commission of Shanghai Municipality(19411950900);Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Grant Support(20171904)

摘要/Abstract

摘要：

目的·探索垂体腺苷酸环化酶激活肽38（pituitary adenylate cyclase-activating polypeptide 38，PACAP38）对心肌细胞辐照敏感性的影响及对急性放射性心肌损伤的潜在防护作用。方法·采用H9C2大鼠心室肌母细胞和C57BL/6J小鼠应用6 MVX射线在体内外构建放射性心肌损伤的辐照模型。在辐照之前对H9C2细胞和C57BL/6J小鼠进行不同剂量的PACAP38预处理。PACAP38在H9C2细胞中的作用浓度为10^-9和10^-7mol/L，在辐照前2 h给予。在小鼠中，分别在辐照前2 h、辐照后24及48 h给予10 μg PACAP38（0.1 μg/μL×100 μL）腹腔注射。体内外放射性心肌损伤辐照模型分为对照组、PACAP38单独处理组、辐照（irradiation， IR）组和PACAP38+IR组。体外实验辐照剂量为2、4、8及12 Gy；体内实验辐照剂量为14 Gy，单次辐照。应用CCK-8实验检测细胞活力；通过克隆形成实验检测辐照敏感性变化；利用苏木精-伊红染色评估小鼠单心脏辐照1个月后心肌组织结构的病理改变。结果·PACAP38预处理H9C2心肌细胞组较单独IR组（12 Gy）的细胞活力显著提高，其中10^-7mol/L PACAP38+IR组和IR组分别为（98.63±2.70）%和（83.67±0.78）%，差异有统计学意义（P=0.000）。2 Gy时的细胞存活率在10^-9和10^-7mol/L PACAP38预处理后从0.53增加至0.63和0.70，放射增敏比在10^-9和10^-7mol/L PACAP38处理组分别为0.95和0.91。通过体内研究发现，PACAP38干预对辐照（14 Gy）后小鼠的心肌组织的病理损伤包括心肌细胞变性、嗜酸性增强、细胞质空泡化、细胞核固缩及心肌纤维扭曲均具有显著的缓解作用。结论·PACAP38干预后可明显降低心肌细胞的辐照敏感性，对急性放射性心肌损伤具有一定的防护作用。

关键词: 垂体腺苷酸环化酶激活肽38, 辐照模型, 放射性心肌损伤, 防护作用

Abstract:

Objective·To explore the effect of pituitary adenylate cyclase-activating polypeptide 38 (PACAP38) on radiosensitivity of myocardial cells and its potential protective effect on acute radiation-induced myocardial injury.

Methods·The radiation-related cardiac injury models were established by using 6 MV X-ray with H9C2 cardiomyocytes and male C57BL/6J mice which were pre-treated with different doses of PACAP38 prior to radiation exposure. H9C2 cells were treated with 10^-9 and 10^-7 mol/L PACAP38 2 h before irradiation. 10 μg of 0.1 μg/μL PACAP38 was administered to C57BL/6J mice intraperitoneally at 2 h before irradiation and additional doses were given at 24 h and 48 h after irradiation. In vivo and in vitro myocardial radiation injury models were divided into control group, PACAP38 group, irradiation group (IR) and PACAP38+IR group. The specific irradiation doses in vitro were 2, 4, 8 and 12 Gy. The specific irradiation dose in vivo were 14 Gy, one fraction irradiation. CCK-8 and clonogenesis assays were used to examine cell viability and radiosensitivity, respectively. Hematoxylin and eosin staining was used to evaluate the pathological changes of myocardial tissue in mice after one-month irradiation.

Results·The cell viability of H9C2 cardiomyocytes pretreated with PACAP38 was significantly higher than that of irradiation (12 Gy) alone [10^-7mol/L PACAP38+IR group vs IR group, (98.63±2.70)% vs (83.67±0.78)%, P=0.000]. The survival fraction at 2 Gy increased from 0.53 to 0.63 and 0.70 after 10^-9 and 10^-7 mol/L PACAP38 pretreatment, and the sensitivity enhancement ratio of 10^-9 and 10^-7 mol/L PACAP38 pretreatment groups were 0.95 and 0.91, respectively. In vivo studies showed that PACAP38 could significantly alleviate the pathological damage of myocardial tissue after irradiation (14 Gy), including cardiomyocyte degeneration, eosinophilic enhancement, cytoplasmic vacuolation, nuclear pyknosis and myocardial fiber distortion.

Conclusion·PACAP38 can significantly reduce the radiosensitivity of myocardial cells and has a certain protective effect on acute radiation-induced myocardial injury.

Key words: pituitary adenylate cyclase-activating polypeptide 38 (PACAP38), irradiation model, radiation-induced myocardial injury, cardioprotectant

中图分类号:

R730

李欢, 易培强, 苏筠, 陈培战, 许赬, 曹璐, 陈佳艺, 李敏. 垂体腺苷酸环化酶激活肽38对急性放射性心肌损伤的防护作用[J]. 上海交通大学学报(医学版), 2021, 41(2): 129-133.

Huan LI, Pei-qiang YI, Jun SU, Pei-zhan CHEN, Cheng XU, Lu CAO, Jia-yi CHEN, Min LI. Protective effect of pituitary adenylate cyclase-activating polypeptide 38 on acute radiation-induced myocardial injury[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(2): 129-133.

图/表 4

图1 PACAP38对辐照诱导的H9C2心肌细胞毒性的防护作用Note: ①P=0.000, compared with the control group; ②P=0.000, ③P=0.014, compared with the IR group.

Fig 1 Protective role of PACAP38 against IR-induced cytotoxicity in H9C2 cells

图2 PACAP38干预对H9C2心肌细胞SF2的影响

Fig 2 Effect of PACAP38 intervention on SF2 of H9C2 cells

表1 PACAP38对H9C2心肌细胞辐照敏感性的影响(xˉ±s)

Tab 1 Effect of PACAP38 on radiosensitivity of H9C2 cells (xˉ±s)

Group	D₀/Gy	D_q/Gy	SF2	SER
Control	1.59±0.01	1.27±0.07	0.53±0.02	1.00
PACAP38(10^-9 mol·L^-1)	1.67±0.03	1.70±0.06	0.63±0.01	0.95±0.02
PACAP38(10^-7 mol·L^-1)	1.76±0.06	1.99±0.11	0.70±0.02	0.91±0.03

图3 PACAP38干预对辐照小鼠心肌组织的保护作用（H-E染色，×400）Note:IR group showed severe cardiac injury characterized by cardiomyocyte degeneration, eosinophilic enhancement (thick arrow), cytoplasmic vacuolization (middle arrow), nuclear condensation and myocardial fiber contorted (thin arrow). Bar=50 μm.

Fig 3 Protective effect of PACAP38 treatment on myocardial tissue after irradiation（H-E staining, ×400）

参考文献 20

1	Delaney G, Jacob S, Featherstone C, et al. The role of radiotherapy in cancer treatment: estimating optimal utilization from a review of evidence-based clinical guidelines[J]. Cancer, 2005, 104(6): 1129-1137.
2	van Nimwegen FA, Schaapveld M, Janus CPM, et al. Cardiovascular disease after Hodgkin lymphoma treatment: 40-year disease risk[J]. JAMA Intern Med, 2015, 175(6): 1007-1017.
3	van Nimwegen FA, Ntentas G, Darby SC, et al. Risk of heart failure in survivors of Hodgkin lymphoma: effects of cardiac exposure to radiation and anthracyclines[J]. Blood, 2017, 129(16): 2257-2265.
4	Clarke M, Collins R, Darby S, et al. Effects of radiotherapy and of differences in the extent of surgery for early breast cancer on local recurrence and 15-year survival: an overview of the randomised trials[J]. Lancet, 2005, 366(9503): 2087-2106.
5	McGale P, Darby SC, Hall P, et al. Incidence of heart disease in 35 000 women treated with radiotherapy for breast cancer in Denmark and Sweden[J]. Radiother Oncol, 2011, 100(2): 167-175.
6	Desai MY, Jellis CL, Kotecha R, et al. Radiation-associated cardiac disease: a practical approach to diagnosis and management[J]. JACC Cardiovasc Imaging, 2018, 11(8): 1132-1149.
7	Vaudry D, Falluel-Morel A, Bourgault S, et al. Pituitary adenylate cyclase-activating polypeptide and its receptors: 20 years after the discovery[J]. Pharmacol Rev, 2009, 61(3): 283-357.
8	Farnham MMJ, Inglott MA, Pilowsky PM. Intrathecal PACAP-38 causes increases in sympathetic nerve activity and heart rate but not blood pressure in the spontaneously hypertensive rat[J]. Am J Physiol Heart Circ Physiol, 2011, 300(1): H214-H222.
9	Khan AM, Li M, Brant E, et al. Renoprotection with pituitary adenylate cyclase-activating polypeptide in cyclosporine A-induced nephrotoxicity[J]. J Investig Med, 2011, 59(5): 793-802.
10	Kouvaris JR, Kouloulias VE, Vlahos LJ. Amifostine: the first selective-target and broad-spectrum radioprotector[J]. Oncologist, 2007, 12(6): 738-747.
11	Demiral AN, Yerebakan O, Simşir V, et al. Amifostine-induced toxic epidermal necrolysis during radiotherapy: a case report[J]. Jpn J Clin Oncol, 2002, 32(11): 477-479.
12	van der Veen SJ, Ghobadi G, de Boer RA, et al. ACE inhibition attenuates radiation-induced cardiopulmonary damage[J]. Radiother Oncol, 2015, 114(1): 96-103.
13	Reglodi D, Vaczy A, Rubio-Beltran E, et al. Protective effects of PACAP in ischemia[J]. J Headache Pain, 2018, 19(1): 19.
14	Racz B, Reglodi D, Horvath G, et al. Protective effect of PACAP against doxorubicin-induced cell death in cardiomyocyte culture[J]. J Mol Neurosci, 2010, 42(3): 419-427.
15	Seaborn T, Masmoudi-Kouli O, Fournier A, et al. Protective effects of pituitary adenylate cyclase-activating polypeptide (PACAP) against apoptosis[J]. Curr Pharm Des, 2011, 17(3): 204-214.
16	Kim EJ, Lee J, Jung YR, et al. Involvement of corin downregulation in ionizing radiation-induced senescence of myocardial cells[J]. Int J Mol Med, 2015, 35(3): 731-738.
17	Dai C, He L, Ma B, et al. Facile nanolization strategy for therapeutic Ganoderma lucidum spore oil to achieve enhanced protection against radiation-induced heart disease[J]. Small, 2019, 15(36): e1902642.
18	Lauk S, Kiszel Z, Buschmann J, et al. Radiation-induced heart disease in rats[J]. Int J Radiat Oncol Biol Phys, 1985, 11(4): 801-808.
19	Hoving S, Heeneman S, Gijbels MJJ, et al. Single-dose and fractionated irradiation promote initiation and progression of atherosclerosis and induce an inflammatory plaque phenotype in ApoE^-/- mice[J]. Int J Radiat Oncol Biol Phys, 2008, 71(3): 848-857.
20	Rezaeyan A, Haddadi GH, Hosseinzadeh M, et al. Radioprotective effects of hesperidin on oxidative damages and histopathological changes induced by X-irradiation in rats heart tissue[J]. J Med Phys, 2016, 41(3): 182-191.

[1]	张凤春, 张硕渊, 陈天恩, 徐海燕, 马越, 刘照南, 徐迎春. 男性肿瘤患者生育力保存认知现状调查[J]. 上海交通大学学报(医学版), 2021, 41(1): 82-88.
[2]	张凤春, 张硕渊, 陈天恩, 徐海燕, 马越, 刘照南 #, 徐迎春 #. 男性肿瘤患者生育力保存认知现状调查[J]. 上海交通大学学报(医学版), 0, (): 1-7.
[3]	滕家俊，聂蔚，高志强，徐建林，孙加源，钟华. 国产经胸壁穿刺诊疗定位系统辅助肺结节穿刺活检的随机对照研究[J]. 上海交通大学学报(医学版), 2020, 40(09): 1218-1221.
[4]	赵伟光，刘志宏. 肿瘤相关成纤维细胞调控肿瘤免疫炎症微环境的研究进展[J]. 上海交通大学学报(医学版), 2020, 40(09): 1288-1293.
[5]	仇晓霞1，金光鑫2，郭艳1，代孟君2，胡嘉娴1，张学彬2. 肿瘤患者上臂植入输液港相关感染发生率及危险因素研究[J]. 上海交通大学学报（医学版）, 2019, 39(10): 1183-.
[6]	张欣，黄锣，陶丹，李丛，谢林，王雨晴，龙斌. 注册于中国临床试验注册中心的放射治疗临床试验特征分析[J]. 上海交通大学学报（医学版）, 2019, 39(7): 789-.
[7]	许开宇 1，魏玉虾 2，王大业 3，孙萍萍 3，周全 3，丁磊 1. 多发性恶性血管球瘤 1例临床病理分析[J]. 上海交通大学学报（医学版）, 2018, 38(12): 1499-.
[8]	赵玲，高润霖，刘强，沈艳莹. 5例甲状腺内胸腺癌临床病理分析[J]. 上海交通大学学报（医学版）, 2018, 38(10): 1272-.
[9]	张义朋 1，黄华艳 1，仰昳婕 1，徐懂懂 1，张可人 1,2，朱亮 1, 2, 3. 受体酪氨酸激酶AXL在肿瘤耐药中的作用研究进展[J]. 上海交通大学学报（医学版）, 2018, 38(7): 819-.
[10]	张凤春 1, 2，马越3，林玉梅 4*，刘照南 3，闫宁宁 1, 2，徐海燕 1，徐迎春 3，王理伟 3. 三级甲等医院肿瘤相关科室医师对于患者生育力保存认知度的调查[J]. 上海交通大学学报（医学版）, 2018, 38(5): 514-.
[11]	梁倩 1, 2，沈瑛 1, 2. 磷酸甘油酸变位酶1在肿瘤发生及发展中的作用[J]. 上海交通大学学报（医学版）, 2018, 38(4): 463-.
[12]	梁倩 1, 2，沈瑛 1, 2. 磷酸甘油酸变位酶1在肿瘤发生及发展中的作用[J]. 上海交通大学学报（医学版）, 0, (): 463-.
[13]	郝亚娟, 刘颖斌, . RNA m6A甲基化修饰酶调控肿瘤的功能及其抑制剂的研究进展[J]. 上海交通大学学报(医学版), 0, (): 1-6.

垂体腺苷酸环化酶激活肽38对急性放射性心肌损伤的防护作用

Protective effect of pituitary adenylate cyclase-activating polypeptide 38 on acute radiation-induced myocardial injury

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 20

相关文章 13

编辑推荐

Metrics