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内质网应激通路相关蛋白ATF4及CHOP在主动脉夹层中的表达以及意义

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目的 探讨主动脉夹层(AD)患者环磷酸腺苷反应元件结合转录因子同源蛋白(CHOP)
和活性转录因子4(ATF4)的表达及其意义。方法 选取2017年3月至2018年4月于武汉大学人民医院确
诊为StanfordⅠ型夹层并行胸主动脉夹层全弓置换患者为AD组(n=13),另选取5例于武汉大学人民医
院接受心脏移植的患者作为Donor组(此5例患者均无大血管疾病,其在心脏移植术中取下的主动脉血
管可作为相对正常主动脉血管组织)。通过Masson染色、间苯二酚染色研究两组患者主动脉血管结构
变化,通过TUNEL染色明确主动脉夹层发病后平滑肌细胞凋亡情况;通过RT-PCR及Western-blot分别
在mRNA水平及蛋白水平检测ATF4及CHOP的变化情况。在动物实验方面,20只SD大鼠随机分为两组:
Control组(n=10)及AD组(n=10)。Control组给予大鼠维持饲料,AD组给予含0.3%的β-异氨基丙腈酯
(BAPN)的大鼠维持饲料进行主动脉夹层动物模型构建。动物模型构建成功后,取两组大鼠主动脉组
织做石蜡包埋,标本切片后处理步骤及检测指标同人体标本。结果 在人体标本中,Masson染色及间苯二
酚染色发现,AD组患者主动脉血管标本胶原沉积较Donor组明显增多,弹力纤维断裂程度也更加严重;
TUNEL凋亡染色发现,AD组患者主动脉血管组织较Donor组正常血管组织细胞凋亡明显增多,差异有统
计学意义(P<0.01);RT-PCR及Western-blot实验发现,AD组患者主动脉血管ATF4、CHOP的表达在
RNA水平及蛋白水平均较Donor组正常主动脉血管明显增高,差异均有统计学意义(P<0.05)。在动物
实验中,Masson染色发现,AD组大鼠主动脉血管较Control组胶原沉积明显增多;间苯二酚染色发现,
AD组大鼠主动脉血管较Control组弹力纤维断裂程度明显加重,Control组基本无弹力纤维断裂;TUNEL
凋亡染色发现,AD组大鼠主动脉血管较Control组细胞凋亡明显增多,差异有统计学意义(P<0.05)。
RT-PCR及Western-blot实验亦发现,AD组大鼠主动脉血管ATF4、CHOP的表达在RNA水平及蛋白水平均
较Control组明显增高,差异均有统计学意义(P<0.05)。结论 CHOP和ATF4与内质网应激关系密切,
CHOP及ATF4在人主动脉夹层标本及主动脉夹层动物模型标本中均显著升高,说明在主动脉夹层中内质
网应激水平较高,这或许是主动脉血管平滑肌细胞发生凋亡的重要原因之一,进而在一定程度上参与主
动脉夹层的发生发展。

Abstract:

Objective To investigate the expression and significance of C/EBP-Homologous protein (CHOP)
and active transcription factor 4 (ATF4) in aortic dissection. Methods Patients who were diagnosed as Stanford type
I with a parallel thoracic aortic dissection from March 2017 to April 2018 were enrolled in the AD group (n=13),
and 5 patients were admitted for heart transplantation. The patients were treated as donors (these 5 patients had no
macrovascular disease, and the aortic vessels removed during heart transplantation were used as relatively normal
aortic vascular tissue). The aortic vascular structure changes were studied by mason staining and resorcinol staining.
The sharp muscle cells after aortic dissection were confirmed by TUNEL staining. ATF4 was detected by RT-PCR
and Western-blot at mRNA and protein levels, respectively. And changes in CHOP. In animal experiments, 20 SD
rats were randomly divided into control group: control group (n=10) and AD group (n=10). Rats in the control group
were given feed, and rats in the AD group were given a 0.3% beta-isopropylaminoglycolate (B APN)-maintained
feed for aortic dissection animal model establishment. After the animal model was successfully constructed, the rat
aortic tissue was implanted for paraffin embedding, and the post-slice treatment steps and detection indexes were
the same as the human specimens. Results In human specimens, Masson staining and resorcinol staining showed
that the collagen deposition in the aortic vascular specimens of the AD group was significantly higher than that of the
Donor group, and the degree of elastic fiber rupture was also more serious. TUNEL apoptosis staining revealed that
the aorta vessels of the AD group were aortic. The apoptosis of the normal vascular tissue was significantly higher
than that in the Donor group (P<0.01). RT-PCR and Western-blot experiments showed that the expression of ATF4
and CHOP in the aortic vessels of the AD group was at the RNA level and protein. The water level was significantly
higher than that of the normal aorta in the Donor group, and the difference was statistically significant (P<0.05). In
animal experiments, Masson staining showed that the aortic vessels of the AD group were significantly increased
compared with the control group. The resorcinol staining showed that the aortic vessels of the AD group were
significantly more severe than the control group. Basic non-elastic fiber rupture; TUNEL apoptosis staining showed
that the apoptosis of aortic vessels in AD group was significantly higher than that in Control group, the difference
was statistically significant (P<0.05). RT-PCR and Western-blot experiments also found that the expression of
ATF4 and CHOP in the aortic vessels of the AD group was significantly higher than that of the Control group (P<0.05).
Conclusion CHOP and ATF4 are closely related to endoplasmic reticulum stress. CHOP and ATF4 are significantly
elevated in human aortic dissection specimens and aortic dissection animal models, indicating higher endoplasmic
reticulum stress levels in the aortic dissection. It may be one of the important causes of apoptosis of aortic vascular
smooth muscle cells, and to some extent participate in the occurrence and development of aortic dissection.

基金项目:

国家自然科学基金(8157020938)

参考文献:

[1] 吴琪,王志维. 血管中膜与主动脉夹层发病机制的研究进展[J].
中国循证心血管医学杂志,2017,9(5):625-8.
[2] He R,Guo DC,Estrera AL,et al. Characterization of the inflammatory
and apoptotic cells in the aortas of patients with ascending
thoracic aortic aneurysms and dissections[J]. J Thorac Cardiovasc
Surg,2006,131(3):671-8.
[3] Hetz C. The unfolded protein response: controlling cell fate
decisions under ER stress and beyond[J]. Nat Rev Mol Cell
Biol,2012,13(2):89-102.
[4] Wang Y,Huang HY,Bian GL,et al. A Functional Variant of SMAD4
Enhances Thoracic Aortic Aneurysm and Dissection Risk through
Promoting Smooth Muscle Cell Apoptosis and Proteoglycan
Degradation[J]. EBioMedicine,2017,21:197-205.
[5] Gawinecka J,Sch?nrath F,von Eckardstein A. Acute aortic
dissection: pathogenesis, risk factors and diagnosis[J]. Swiss Med
Wkly,2017,147:w14489.
[6] Hetz C,Papa FR. The Unfolded Protein Response and Cell Fate
Control[J]. Mol Cell,2018,69(2):169-81.
[7] Han J,Back SH,Hur J,et al. ER-stress-induced transcriptional
regulation increases protein synthesis leading to cell death[J]. Nat
Cell Biol,2013,15(5):481-90.
[8] Inoue Y,Kawachi S,Ohkubo T,et al. The CDK inhibitor p21 is a
novel target gene of ATF4 and contributes to cell survival under ER
stress[J]. FEBS Lett,2017,591(21):3682-91.
[9] Fu HY,Okada KI,Liao Y,et al. Ablation of C/EBP Homologous Protein
Attenuates Endoplasmic Reticulum-Mediated Apoptosis and Cardiac
Dysfunction Induced by Pressure Overload[J]. Circulation,2010,122
(4):361-9.
[10] Bruhat A,Jousse C,Carraro V,et al. Amino Acids Control Mammalian
Gene Transcription: Activating Transcription Factor 2 Is Essential for
the Amino Acid, Responsiveness of the CHOP Promoter[J]. Mol Cell
Biol,2000,20(19):7192-204.
[11] Marciniak SJ,Yun CY,Oyadomari S,et al. CHOP induces death by
promoting protein synthesis and oxidation in the stressed endoplasmic
reticulum[J]. Genes Dev,2004,18(24): 3066-77.
[12] Morse E,Schroth J,You YH,et al. TRB3 is stimulated in diabetic
kidneys, regulated by the ER stress marker CHOP, and is a
suppressor of podocyte MCP-1[J]. Am J Physiol Renal Physiol,
2010,299(5):F965-72.
[13] Urra H,Dufey E,Lisbona F,et al. When ER stress reaches a dead
end[J]. Biochim Biophys Acta,2013,1833(12):3507-17.
[14] Bruchmann A,Roller C,Walther TV,et al. Bcl-2 associated athanogene
5 (Bag5) is overexpressed in prostate cancer and inhibits ER-stress
induced apoptosis[J]. BMC Cancer,2013,13:96.
[15] Wang XZ,Kuroda M,Sok J,et al. Identification of novel stress-induced
genes downstream of chop[J]. EMBO J,1998,17(13):3619-30.
[16] Silva RM,Ries V,Oo TF,et al. CHOP/GADD153 is a mediator
of apoptotic death in substantia nigra dopamine neurons in an
in vivo neurotoxin model of parkinsonism[J]. J Neurochem,
2010,95(4):974-86.
[17] 程宝山,王爱玲. 脉压差与急性主动脉夹层院内预后关系的研究
[J]. 中国心血管病研究,2017,15(2):146-9.
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