The Journal of Practical Medicine >

2024 , Vol. 40 >Issue 5: 639 - 645

DOI: https://doi.org/10.3969/j.issn.1006-5725.2024.05.009

Basic Research

Experimental study on treatment of severe limb ischemia with Ad⁃hVEGF⁃hHGF gene

  • Rui ZHONG ,
  • Jianing WANG ,
  • Lei ZHANG ,
  • Lingyun GUO ,
  • Jianye YANG ,
  • Fei ZHENG ,
  • Yuwen YAN ,
  • Danli YU ,
  • Liguo. TAN
Expand
  • Department of Cardiology，People's Hospital Affiliated to Hubei University of Medicine，Shiyan 442000，China

Received date: 2023-06-20

  Online published: 2024-03-26

Fold

Abstract

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue. Methods 84 Kunming mice were randomly divided into sham group， control group， VEGF group， HGF group and VEGF+HGF group， and the left lower limb ischemia model was established. The blood supply of ischemic tissue was observed by rheometer， and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA. Immunohistochemical staining was used to detect angiogenesis （CD31， SMA） in ischemic tissues. Safety was assessed by side effects during treatment in mice. Results After the successful modeling， the blood flow velocity of the left lower limb in each group decreased significantly. On the 7th day after operation， the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation （P < 0.05）， and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups （P < 0.05）. On the 28th day after operation， the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized， the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups， and both VEGF group and HGF group were significantly better than the control group （P < 0.05）. On the 7th， 14th， and 28th days following surgery， HGF and VEGF protein levels in the Ad-HGF， Ad-VEGF， and Ad-VEGF-HGF groups were substantially greater than those in the control group （P < 0.05）. The expression level in the Ad-VEGF-HGF group peaked on the 14th day （all P < 0.001） and subsequently declined to preoperative levels on the 28th day after operation. Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level， encouraging angiogenesis after lower limb ischemia， increasing blood flow， and improving lower limb circulation.

Key words： vascular endothelial growth factor; stem cell growth factor; lower limb ischemia; adenovirus vector

Cite this article

Rui ZHONG , Jianing WANG , Lei ZHANG , Lingyun GUO , Jianye YANG , Fei ZHENG , Yuwen YAN , Danli YU , Liguo. TAN . Experimental study on treatment of severe limb ischemia with Ad⁃hVEGF⁃hHGF gene[J]. The Journal of Practical Medicine, 2024 , 40(5) : 639 -645 . DOI: 10.3969/j.issn.1006-5725.2024.05.009

References

1 王书凡，赫玉香，程小龙，等. 杜鹃花总黄酮通过激活大鼠脑基底动脉血管内皮细胞TRPV4、Kca3.1和Kca2.3通道改善缺血性脑损伤［J］. 皖南医学院学报，2020，39（6）：516-520. doi:10.3969/j.issn.1002-0217.2020.06.002
2 王爱红，石鸿雁，许樟荣. 基于临床症状和体征评估糖尿病足溃疡肢体缺血严重程度的价值［J］. 中华糖尿病杂志，2020，12（8）：585-588. doi:10.3760/cma.j.cn115791-20200506-00268
3 梁永红，冯超，梁顺添，等. 血管腔内介入治疗下肢严重缺血后急性肾损伤危险因素［J］. 中国介入影像与治疗学，2021，18（2）：91-94.
4 REIS P E， DE CARVALHO L P， YASUMURA E， et al. Impact of angiogenic therapy in the treatment of critical lower limb ischemia in an animal model［J］. Vasc Endovascular Surg， 2014，48（3）：207-216. doi:10.1177/1538574413518119
5 李志娟，庄百溪，马鲁波. 慢性威胁肢体性缺血中医药介入治疗研究进展［J］. 中西医结合心脑血管病杂志，2021，19（6）：956-958. doi:10.12102/j.issn.1672-1349.2021.06.016
6 血管外科学会、欧洲血管外科学会和世界血管学会联盟全球血管指南编写小组. 慢性肢体威胁性缺血治疗的全球血管指南（全译）［J］. 中华血管外科杂志，2021，6（Z1）：1-108，F3.
7 杨川，郭发才，周栋. 重症下肢缺血治疗的研究进展［J］.重庆医学，2015，44（4）：544-547.
8 WATANABE M， NISHIKAWAJI Y， KAWAKAMI H， et al. Adenovirus biology，recombinant adenovirus，and adenovirus usage in gene therapy［J］.Viruses，2021，13（12）：2502. doi:10.3390/v13122502
9 SAKURAI F， TACHIBANA M， MIZUGUCHI H. Adenovirus vector-based vaccine for infectious diseases［J］. Drug Metab Pharmacokinet， 2022，42：100432. doi:10.1016/j.dmpk.2021.100432
10 杨宇，董继红，张荣荣，等. 年龄相关性黄斑变性患者抗VEGF治疗视力预后的影响因素研究［J］. 皖南医学院学报，2022，41（1）：36-39. doi:10.3969/j.issn.1002-0217.2022.01.010
11 OZGUIR E， HEIDENREICH A， DAGTEKIN O， et al. Distribution of EphB4 and EphrinB2 in normal and malignant urogenitaltissue［J］. Urol Oncol，2011，29（ 1）：78-84. doi:10.1016/j.urolonc.2008.12.020
12 VIJAYAKUMAR A， TIWARI R， KUMAR PRABHUSWAMY V.Throm-boangiitis obliterans （Buerger' s disease）-current practices［J］. Int J Inflam，2013，20（13）：1569.
13 SHIGEMATSU H， YASUDA K， IWAI T， et al. Randomized，double-blind， placebo-controlled clinical trial of hepato-cyte growth factor plasmid for critical limb ischemia［J］. Gene Ther，2010，17（9）：1152-1161. doi:10.1038/gt.2010.51
14 MORISHITA R， MAKINO H， AOKI M，et al. Phase I/IIa clinicaltrial of therapeutic angiogenesis using hepatocyte growthfactor gene transfer to treat critical limb ischemia［J］. Arterioscler Throm Vas Biol， 2011，31（3）：713-720. doi:10.1161/atvbaha.110.219550
15 FARBER A， EBERHARDT R T. The current state of critical limb ischemia： a systematic review［J］. JAMA Surg， 2016， 151（11）： 1070-1077. doi:10.1001/jamasurg.2016.2018
16 FARD B， DIJKSTRA P U， NEDA Study Group， et al. Mortality， reamputation， and preoperative comorbidities in patients undergoing dysvascular lower limb amputation［J］. Ann Vasc Surg， 2020， 64： 228-238. doi:10.1016/j.avsg.2019.09.010
17 FABIANI I， CALOGERO E， PUGLIESE N R， et al. Critical limb ischemia： a practical up-to-date review［J］. Angiology， 2018， 69（6）： 465-474. doi:10.1177/0003319717739387
18 HAGHIGHAT L， IONESCU C N， REGAN C J， et al. Review of the current basic science strategies to treat critical limb ischemia［J］. Vasc Endovascular Surg， 2019， 53（4）： 316-324. doi:10.1177/1538574419831489
19 郭平凡. 下肢慢性缺血性疾病的诊治［J］. 中国血管外科杂志（电子版），2021，13（4）：299-301，305.
20 JOHNSON T， ZHAO L， MANUEL G， et al. Approaches to therapeutic angiogenesis for ischemic heart disease［J］. J Mol Med， 2019， 97（2）： 141-151. doi:10.1007/s00109-018-1729-3
21 YL?-HERTTUALA S. Gene therapy of critical limb ischemia enters clinical use［J］. Mol Ther， 2019， 27（12）： 2053. doi:10.1016/j.ymthe.2019.11.001
22 M?KINEN K， MANNINEN H， HEDMAN M， et al. Increased vascularity detected by digital subtraction angiography after VEGF gene transfer to human lower limb artery： a randomized， placebo-controlled， double-blinded phase Ⅱ study［J］. Mol Ther， 2002， 6（1）： 127-133. doi:10.1006/mthe.2002.0638
23 BAR? P， ANTKIEWICZ M， ?LIWA B， et al. Double VEGF/HGF gene therapy in critical limb ischemia complicated by diabetes mellitus［J］. J Cardiovasc Transl Res， 2021，14： 409-415. doi:10.1007/s12265-020-10066-9
24 李存娣，曹伟娅，王健. 血管内皮生长因子受体与肝细胞癌侵袭转移的关系［J］. 临床肝胆病杂志，2019，35（7）：1616-1620. doi:10.3969/j.issn.1001-5256.2019.07.042
25 RIAUD M， HILAIRET G， SINDJI L， et al. Pharmacology active microcarriers delivering HGF associated with extracellular vesicles for myocardial repair［J］. Eur J Pharm Biopharm， 2021， 169： 268-279. doi:10.1016/j.ejpb.2021.10.018
26 ROSENGART T K， LEE L Y， PATEL S R， et al.Angiogenesis Gene Therapy Phase IAssessment of Direct Intramyocardial Administration of an Adenovirus VectorExpressing VEGF12i cDNA to Individuals With Clinically Significant SevereCoronary Artery Disease［J］. Circulation， 1999，100：468-474. doi:10.1161/01.cir.100.5.468
Options
Outlines

/

〈 〉