基于LncRNA NANCI-NKX2.1信号通路探讨清肺养阴活血方保护放射性肺损伤的作用研究
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摘要
目的探讨清肺养阴活血方对放射性肺损伤小鼠模型的保护作用以及对长非编码RNA NANCI-NKX2.1信号通路的影响。方法将C57BL/6小鼠分为空白对照组(A组)、中药对照组(B组)、模型对照组(C组)和中药模型组(D组),每组各16只。建模后次日开始给予清肺养阴活血方干预。给药10周后,将动物颈椎脱臼处死,观察肺组织病理学变化,并进行H&E染色和Masson染色。采用qRT-PCR法和Western blot法检测长链非编码RNA NANCI和NKX2.1 mRNA和蛋白的表达水平。结果 A组和B组动物体重变化率基本一致(P>0.05)。直至实验结束时,虽然D组动物体重一直低于A组和B组动物(P<0.05);但是自给药7 d后,D组动物体重逐渐高于C组动物(P<0.05)。经H&E染色和Masson染色,A组和B组肺组织结构完整,C组可见明显的炎性细胞浸润,大量胶原纤维沉着,而D组动物肺组织病理学变化情况较C组明显改善。D组动物肺组织损伤评分为(3.875±1.746)分,明显高于A组和B组动物评分,但是低于C组动物评分(P<0.05)。另外,D组动物肺组织lncRNA NANCI、NKX2.1 mRNA和蛋白相对表达量明显低于A组和B组动物表达量,但是高于C组动物表达量(P<0.05)。且与肺组织损伤评分呈负相关性(r=-0.510,-0.786,P<0.05)。结论清肺养阴活血方可能通过正性调控lncRNA NANCI-NKX2.1信号通路保护放射性肺组织损伤。
OBJECTIVE To discuss the effect of Qingfei-yangyin-huoxue recipe on radiation-induced pulmonary injury by regulating the lncRNA NANCI-NKX2. 1 pathway. METHODS The C57 BL/6 mice were randomly divided into normal control group( A group,n = 16),drug only group( B group,n = 16),model group( C group,n = 16) and drug model group( D group,n = 16). After exposure to Qingfei-yangyin-huoxue recipe for 10 w,the pathological change of lung tissue was examined by H&E and Masson staining. The expressions of lncRNA NANCI and NKX2. 1 mRNA in lung tissues were detected by qRTPCR. And the NKX2. 1 protein expressions were detected by Western blot. RESULTS The mean animal weight in D groups was less than A and B group,but more than C group after treatment of 7 d( P < 0. 05). There were marked interstitial edema and inflammatory cells,fibrocytes accumulation in C group but not in A and B groups by H&E and Masson stain. The alveolitis and fibrosis changes in D group were better than C group. And the mean radiation-induced pulmonary injury score in D group was( 3. 875 ± 1. 746),which was less than C group,but more than A and B groups( P < 0. 05). The expression of lncRNA NANCI and NKX2. 1 in D group was higher than C group,but lower than A and B groups( P < 0. 05). Besides,the radiation-induced pulmonary injury score was negative related with lncRNA NANCI and NKX2. 1( r =-0. 510,-0. 786,P < 0. 05). CONCLUSION There are significant evidences that Qingfei-yangyin-huoxue recipe could protect radiation-induced pulmonary injury by up-regulation lncRNA NANCI-NKX2. 1 pathway.
OBJECTIVE To discuss the effect of Qingfei-yangyin-huoxue recipe on radiation-induced pulmonary injury by regulating the lncRNA NANCI-NKX2. 1 pathway. METHODS The C57 BL/6 mice were randomly divided into normal control group( A group,n = 16),drug only group( B group,n = 16),model group( C group,n = 16) and drug model group( D group,n = 16). After exposure to Qingfei-yangyin-huoxue recipe for 10 w,the pathological change of lung tissue was examined by H&E and Masson staining. The expressions of lncRNA NANCI and NKX2. 1 mRNA in lung tissues were detected by qRTPCR. And the NKX2. 1 protein expressions were detected by Western blot. RESULTS The mean animal weight in D groups was less than A and B group,but more than C group after treatment of 7 d( P < 0. 05). There were marked interstitial edema and inflammatory cells,fibrocytes accumulation in C group but not in A and B groups by H&E and Masson stain. The alveolitis and fibrosis changes in D group were better than C group. And the mean radiation-induced pulmonary injury score in D group was( 3. 875 ± 1. 746),which was less than C group,but more than A and B groups( P < 0. 05). The expression of lncRNA NANCI and NKX2. 1 in D group was higher than C group,but lower than A and B groups( P < 0. 05). Besides,the radiation-induced pulmonary injury score was negative related with lncRNA NANCI and NKX2. 1( r =-0. 510,-0. 786,P < 0. 05). CONCLUSION There are significant evidences that Qingfei-yangyin-huoxue recipe could protect radiation-induced pulmonary injury by up-regulation lncRNA NANCI-NKX2. 1 pathway.
引文
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[20]SUN F,ZHANG J X,YANG C Y,et al.The genetic characteristics of congenital hypothyroidism in China by comprehensive screening of 21 candidate genes[J].Eur J Endocrinol,2018,178(6):623-633.
[2]LIU F,WANG C,HU T,et al.S-1-based concurrent chemoradiotherapy in the treatment of locally advanced non-small cell lungcancer:a systematic review and Meta-analysis protocol[J].Medicine(Baltimore),2018,97(15):e0397.
[3]YAZBECK V Y,VILLARUZ L,HALEY M,et al.Management of normal tissue toxicity associated with chemoradiation(primary skin,esophagusandlung)[J].Cancer J,2013,19(3):231-237.
[4]ZHAO J,YORKE E D,LI L,et al.Simple factors associatedwithradiation-inducedlung toxicity after stereotactic body radiation therapy of the thorax:apooledanalysis of 88 studies[J].Int J Radiat Oncol Biol Phys,2016,95(5):1357-1366.
[5]YOU F M,ZHENG C,ZHU J,et al.Study of treatment of radioactive-induced lung injury[J].J Tradit Chin Med(中医杂志),2015,60(21):1819-1821.
[6]YANG X X,TIAN J H,ZHANG J,et al.Clinical study and mechanism of Yiqi yangyin methodin treatment of cancer[J].Acta Univ Tradit Med Sin Pharmacol Shanghai(上海中医药大学学报),2015,29(4):87-90.
[7]HERRIGES M J,SWARR D T,MORLEY M P,et al.Long noncoding RNAs are spatially correlated with transcription factors and regulate lung development[J].Genes Dev,2014,28(12):1363-1379.
[8]ARKOVITZ M S,GARCIA V F,SZABO C,et al.Decreased pulmonary compliance is an early indicator of pulmonary oxygen injury[J].J Surg Res,1997,67(2):193-198.
[9]SHI Y K,SUN Y,YU J M,et al.China experts consensus on the diagnosis and treatment of advanced stage primary lung cancer(2016 version)[J].Chin J Lung Cancer(中国肺癌杂志),2016,19(1):1-15.
[10]SENAN S,RUSTHOVEN C G,SLOTMAN B J,et al.Progress in radiotherapy for regional and oligometastatic disease in 2017[J].J Thorac Oncol,2018,13(4):488-496.
[11]SIMONE C B 2ND.Thoracic radiation normal tissue injury[J].Semin Radiat Oncol,2017,27(4):370-377.
[12]LIU R F,WEI S H,ZHANG Q N,et al.Influence of different radio-chemotherapy modes for incidence of radiation pneumonitis in patients with lung cancer[J].Cancer Res Prev Treat(肿瘤防治研究),2018,46(6):5.
[13]LU F L,CHANG L,JIN H Y,et al.Risk factors for radioactive pneumonia[J].Mod Oncol,2015,23(11):1606-1609.
[14]LI C,LEI Z,LU H D,et al.Initial analysis on correlation between lung paralysis,astrophic lung and radiation-induced lung injury[J].Chin Arch Tradit Chin Med(中华中医药学刊),2018,37(1):103-105.
[15]LI C,XIAO X,LEI Z,et al.Analysis of radiation-induced pulmonary injury based on theory of collaterals disease[J].Chin Arch Tradit Chin Med(中华中医药学刊),2017,36(8):2003-2006.
[16]DONG G T,QI X,LI Z,et al.Mechanism debate of warm disease‘Yangyin qingfei huoxue method’on preventing and controlling of radioactive lung injury[J].J Liaoning Univ Tradit Chin Med(辽宁中医药大学学报),2017,19(7):78-81.
[17]ZHANG Z M,PANG D.Research progress of long non-coding RNA regulates cancer cell signaling pathways[J].Pract Oncol J,2018,33(2):184-188.
[18]ZHANG Y,CHENG H P,BAO T P,et al.Expression of long non-coding RNA NANCI in lung tissues of neonatal mice with hyperoxia-induced lung injury and its regulatory effect on NKX2.1[J].Chin J Contemp Pediatr(中国当代儿科杂志),2017,19(2):215-221.
[19]HERRIGE M J,TISCHFIELD D J,CUI Z,et al.The NANCINkx2.1 gene duplex buffers Nkx2.1 expression to maintain lung development and homeostasis[J].Genes Dev,2017,31(9):889-903.
[20]SUN F,ZHANG J X,YANG C Y,et al.The genetic characteristics of congenital hypothyroidism in China by comprehensive screening of 21 candidate genes[J].Eur J Endocrinol,2018,178(6):623-633.