QUANTITATIVE IMMUNOHISTOCHEMICAL EVALUATION OF THE EFFICACY OF RADIOFREQUENCY PULMONARY ARTERY DENERVATION

Radiofrequency pulmonary artery denervation (RFA-PA) is increasingly used by cardiac surgeons worldwide for the simultaneous surgical correction of cardiac defects and arrhythmias in patients with severe pulmonary hypertension. The aim of this study was to morphologically evaluate the efficiency of pulmonary artery radiofrequency denervation using immunohistochemical staining of ganglionic nerve plexus and autonomic nerve fibres with antibodies against S-100 protein. The study material consisted of 30 lung trunks from people who had died of extra-cardiac pathology, aged between 31 and 65 years (section material obtained at routine autopsies). Immunohistochemical staining was then performed on Leica Bond MAX immunostainer with polyclonal antibody against S-100 protein. The relative area of nerve fibres (Sotn.) was used as a statistical value as the ratio of the average area of stained nerve fibres in the field of view to the area of the whole field of view. Pulmonary nerve fibres after staining for S-100 protein were detected mainly along the left lateral edge of the pulmonary trunk, median Sotn. = 5.72% (95% DI 4.27-8.95). A small number of fine nerve fibres were found in the anterior and posterior walls of the lung trunk, Sotn = 0.99% (95% CI 0.48-0.93), p < 0.0000. In sections of the pulmonary trunk in areas subjected to radiofrequency denervation, nerve fibres did not stain for S-100 protein. This is due to the fact that thermal coagulation of the proteins resulted in the loss of specific three-dimensional structure and antigenic properties, indicating the destruction of autonomic nerve fibres of the pulmonary trunk. Quantitative immunohistochemical evaluation of nerve fibre damage using antibodies against S-100 protein allows the efficiency of RFA of DLA to be estimated.

1. Ablyatsiya ganglionarnykh spletenii legochnoi arterii pri khirurgicheskom lechenii poro-kov mitral'nogo klapana u patsientov s vysokoi legochnoi gipertenziei / Bogachev–Prokop'ev A.V., Zheleznev S.I., Afanas'ev A.V. et all. // Patologiya krovoobrashcheniya i kardiokhirurgiya. 2015. vol. 19, № 4. S. 19–25.

2. Goncharov P.N., Sakibaev K.Sh., Goncharov N.I. Illyustratsii k innervatsii serdtsa: pamyati Vladimira Petrovicha Vorob'eva (1876–1937) [Illustrations for the innervation of the heart: in memory of Vladimir Petrovich Vorobyov] Morfologiya. 2008. vol. 133, № 2. pp. 34.

3. Guseva I.A., Shabalin A.V., Kazarinova Yu.L. Svyaz' osobennostei innervatsii serdtsa s mekhanizmami razvitiya supraventrikulyarnykh takhikardii i s posledstviyami radiochastotnoi ablyatsii aritmogennykh ochagov [The relationship of the features of cardiac innervation with the mechanisms of development of supraventricular tachycardia and with the consequences of radiofrequency ablation of arrhythmogenic foci] Vestnik aritmologii. 2001. № 22. pp. 69–74.

4. Denervatsiya legochnogo stvola i ust'ev legochnykh arterii u patsientov s khirurgicheskoi korrektsiei patologii mitral'nogo klapana na fone legochnoi gipertenzii / Trofimov N.A., Medvedev A.P., Dragunov A.G. et all. // Al'manakh klinicheskoi meditsiny. 2017. vol. 45b, № 3. S. 192–199.

5. Primenenie immunogistokhimicheskoi reaktsii na belok PGP 9.5 dlya izucheniya innerva-tsii serdtsa krysy i cheloveka / Korzhevskii D.E., Sukhorukova E.G., Petrova E.S., Tsukanova A.F., Chumasov E.I. // Morfologiya. 2013. vol. 143, № 3. S. 077–080.

6. Radiochastotnaya denervatsiya legochnoi arterii pri khirurgicheskoi korrektsii displa-sticheskikh porokov mitral'nogo klapana s vysokoi legochnoi gipertenziei / Zheleznev S.I., Demidov D.P., Afanas'ev A.V. et all. // Rossiiskii kardiologicheskii zhurnal. № 11 (139). 2016. S. 70–72.

7. Rudenko V.A., Feschenko D.A. Shanoyan A.S. Endovaskulyarnye metody denervatsii legoch-noi arterii v lechenii patsientov s legochnoi gipertenziei: posobie / Pod red. Drapkinoi O.M. M.: FGBU «Natsional'nyi meditsinskii issledovatel'skii tsentr terapii i profilaktiki medi-tsiny Ministerstva zdravookhraneniya Rossiiskoi Federatsii», 2020. 66 s.

8. Chumasov E.I., Petrova E.S., Korzhevskii D.E. Opyt izucheniya innervatsii serdtsa s pomoshch'yu immunogistokhimicheskikh metodov. Morfologicheskie shkoly segodnya. Mater. Vseros. nauchno–prakticheskaya konferentsiya s mezhdunarodnym uchastiem [The experience of studying the innervation of the heart using immunohistochemical methods] Voronezh, 2022. pp. 360–363.

9. Shcherbakova M.N., Gadzhieva F.G. Osobennosti innervatsii serdtsa. Vesennie anatomicheskie chteniya [Features of the innervation of the heart. Spring Anatomical readings] Sbornik statei nauchno–prakticheskoi konferentsii, posvyashchennoi pamyati dotsenta M.A. Kolesova. Grodno, 2016. pp. 254–258.

10. Effektivnost' i bezopasnost' radiochastotnoi ablyatsii legochnogo stvola i ust'ev le-gochnykh arterii pri vysokoi legochnoi gipertenzii (po rezul'tatam gistologicheskogo issledovaniya autopsiinogo materiala) / Trofimov N.A., Babokin V.E., Medvedev A.P. et all. // Trudnyi pa-tsient. 2021. vol. 19b, № 2. S. 6–13. DOI: 10.224412/2074-1995-2021-2-6-13.

11. Chen S.L., Zhang Y.J., Zhou L., et all. Percutaneous pulmonary artery denervation completely abolishes experimental pulmonary arterial hypertension in vivo EuroIntervention, 2013, № 22. pp. 269–276. DOI: 10.4244/EIJV9I2A43.

12. Condori Leandro H.I., Vakhrushev A.D., Goncharova N.S., Korobchenko L.E., Koshevaya E.G., Mitrofanova L.B., Andreeva E.M., Moiseeva O.M., Lebedev D.S., Mikhaylov E.N. Stimulation Mapping of the Pulmonary Artery for Denervation Procedures: an Experimental Study J Cardiovasc Transl Res., 2021, vol. 14, № 3. pp. 546–555. DOI: 10.1007/s12265-020-10079-4.

13. Feshchenko D.A., Rudenko B.A., Shanoyan A.S., Drapkina O.M., Kontsevaya A.V., Gavrilova N.E., Shukurov F.B., Vasil’ev D.K. Pulmonary Artery Denervation for Pulmonary Hypertension: stages of development and clinical experience Russian Journal of Cardiology, 2019, vol. 24, № 12. pp. 162–168. (In Russ.) DOI: 10.15829/1560-4071-2019-12-162-168.

14. George L. Kumar, Lars Rudbeck. Immunohistochemical Staining Methods Education Guide. DAKO, 2011, pp. 224

15. Trofimov N.A., Rodionov A.L., Egorov D.V., Surkova T.V., Nikolsky A.V. Histological justification for the need of radiofrequency ablation of pulmonary arteries in patients with high–grade secondary pulmonary hypertension Sovremennye tehnologii v medicine, 2021, vol. 13, № 6. pp. 56–64. DOI: 10.17691/stm2021.13.6.06.

16. Zhou L., Zhang J., Jiang X.M., et all. Pulmonary artery denervation attenuates pulmonary arterial remodeling in dogs with pulmonary arterial hypertension induced by dehydrogenized monocrotaline J. Am. Coll. Cardiol Intv., 2015, № 8. pp. 2013–2023. DOI: 10.1016/j. jcin.2015.09.015.