Immunoadhesive regulation using multiphytoadaptogene for spontaneous hepatocarcinomas prevention

The purpose of this investigation was to evaluate the efficacy of correcting the LFA-1 and Mac-1 leukocyte integrins expression on peripheral blood cells as well as IL-6 and IL-10 serum levels using liquid form multiphytoadaptogene preventive application for hepatocarcinomas incidence suppression and life-span increase of CBA inbreed mice.

Materials and methods. The study was carried out on 439 males of inbred CBA mouse strain (subline CBA/LacY). The experimental mice received 10 % liquid form multiphytoadaptogene complex (MPAC) solution in drinking water during the first month of life including the final time period of liver differentiation (preventive administration). MPAC is the standardized herbal formula composed of 40 plant extracts components including adaptogenes Panax ginseng, Eleutherococcus senticosus, Rhodiola rosea, as well as compounds with phenolic structure (flavonoids, triterpene glycosides, etc). Anti-mutagenic, anti-oxidant, immunomodifying activities of MPAC were demonstrated. The CD11a and CD11b antigens expressions on peripheral blood cells were analyzed by indirect immunofluorescence reaction, serum cytokines IL-6 and IL-10 concentrations were determined by enzyme-linked immunosorbent assay as well as the liver tissue morphology were analyzed at the age of 4, 8, 22 months. Tumor incidence and volumes were evaluated at the age of 8 and 22 months. Motor activity and physical status (body weight, coat state) were estimated in the ontogenesis. The average life-span and survival median was analyzed by the Kaplan–Meier method. Statistical analysis was performed with the program STATISTICA 6.0.

Results. Inhibited expression of LFA-1 and Mac-1 leukocyte integrins on peripheral blood cells as well as elevated IL-6 and IL-10 serum levels, high incidence of liver tumors (100 % of mice-males), their high number and big sizes in the late period of postnatal ontogenesis in CBA high-cancer inbrede mice, also as life-span not reaching two years was demonstrated. At the same time plural tumor-infiltrating limphocytes were not found. Short-term MPAC preventive administration (during the first month of mouse life occupying the final differentiation period of liver tissue genetically predisposal to hepatoms incidence) revealed the long-term heterotypic adhesion molecules leukocyte integrins LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) up-regulated expression, providing the contact interactions of immune effectors and cancer cells. The events shown is able to enhance the anti-tumor activity of immune reactions including spontaneous hepatocarcinomas lymphocytes infiltration and destruction of tumor tissue. As a result, the hereditary tumors incidence, their number and sizes were reduced as well as life-span and life quality of animals were improved.

Conclusion. Up-regulated leukocyte integrins expression on peripheral blood cells, reduced IL-6 and IL-10 serum levels accompanied by tumors lymphocyte infiltration and destruction using MPAC preventive administration is essential for tumor incidence down-regulation. The herbal formula with multiple components is capable to controle anti-tumor immune reactions, as well as the life-span and life quality of high-cancer animals. Hence, the regulation of adhesion violations involved in tumor incidence can be cancer preventive.

1. Hassan A.M., Abdel-Aziem S.H., El-Nekeety A.A., Abdel-Wahhab M.A. Panax ginseng extract modulates oxidative stress, DNA fragmentation and up-regulate gene expression in rats sub chronically treated with aflatoxin B1 and fumonisin B1. Cytotechnology 2015;67(5):861–71. DOI: 10.1007/s10616-014-9726-z. PMID: 24748134.

2. Panwar M., Kumar M., Samarth R., Kumar A. Evaluation of chemopreventive action and antimutagenic effect of the standardized Panax ginseng extract, EFLA400, in Swiss albino mice. Phytother Res 2005;19(1):65–71. DOI: 10.1002/ptr.1584. PMID: 15799001.

3. Xie J.T., Shao Z.H., Vanden Hoek T.L. et al. Antioxidant effects of ginsenoside Re in cardiomyocytes. Eur J Pharmacol 2006;532(3):201–7. PMID: 16497296.

4. Kim N.D., Pokharel Y.R., Kang K.W. Ginsenoside Rd enhances glutathione levels in H4IIE cells via NF-kappaB-dependent gamma-glutamylcysteine ligase induction. Pharmazie 2007;62(12):933–6. PMID: 18214346.

5. Goldberg E.D., Razina T.G., Zueva E.P. et al. Plants in complex therapy of tumors. М.: Izdatel’stvо RAMN = RAMS publishing house, 2008. 432 p. (In Russ.)

6. Jin Y., Kotakadi V.S., Ying L. et al. American ginseng suppresses inflammation and DNA damage associated with mouse colitis. Carcinogenesis 2008;29(12):2351–9. DOI: 10.1093/carcin/bgn211. PMID: 18802031.

7. Bocharova O.A. Adhesive concept in biology of malignant growth. Patologicheskaya fiziologiya i ehksperimentalnaya terapiya = Pathological Physiology and Experimental Therapy 2014;2:87–90 (In Russ.)

8. Allen S., Moran N. Cell Adhesion Molecules: Therapeutic Targets for Inhibition of Inflammatory States. Semin Thromb Hemost 2015;41(6):563–71. DOI: 10.1055/s-0035-1556588. PMID: 26322694.

9. Bocharova O.A., Modyanova E.A. Hepatocyte cell-cell contacts alterations during ontogenesis of inbred strains of mice with high and low frequency of spontaneous hepatomas. Ontogenez = Ontogenesis 1982;13(4):427–29 (In Russ.)

10. Modyanova E.A., Bocharova O.A., Malenkov A.G. The kontaktins-chalones preventive action on spontaneous carcinogenesis in mice inbred strains. Ehksperimentalnaya onkologiya = Experimental oncology 1983;5(3):39–42 (In Russ.)

11. Bocharova O.A., Serebryakova R.V., Bodrova N.B. Rhodiola rosea preventive effect on spontaneous tumorigenesis in the liver in high-cancer inbred mice. Vestnik RAMN = Bulletin of RAMS 1994;5:41–3 (In Russ.)

12. Zhang H.M., Chen S.W., Xie C.G. Effects and mechanism of ShenQi compound recipe on inflammation marker in GK rats. Zhong Yao Cai 2006;29(3):249–53. PMID: 16850723.

13. Shejchenko O.P., Bocharova O.A., Krapivkin B.A. et al. Study of complex phytoadaptogen by HPLC method. Voprosy biologicheskoy, meditsinskoy i farmstsevticheskoy khimii = Connected Questions of Biological, Medical and Pharmaceutical Chemistry 2012;10:52–9 (In Russ.)

14. Karpova R.V., Shevchenko V.E., Bocharov E.V. et al. Possibility of using high performance liquid chromatography coupled with tandem mass spectrometry for qualitative and quantitative determination of biologically active substances in the ginseng phytoextracts. Rossysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2016;15(2):36–46. (In Russ.)

15. Kurennaya O.N., Karpova R.V., Bocharova O.P. et al. Multiphytoadaptogen antimutagenesis in yeast cells of Saccharomyces. Genetika = Russian Journal of Genetiсs 2013;49(12):1364–69. (In Russ.)

16. Bochkov N.P., Bocharova O.A., Aksyonov A.A. et al. The frequency of chromosomal aberrations in lymphocytes of patients with benign prostatic hyperplasia. Meditsinskaya genetika = Medical Genetics 2005;4(1):15–9 (In Russ.)

17. Pozharitskaya M.M., Bocharova O.A., Chekalina T.L., Voronin V.F. Modern aspects of pathogenesis and treatment of the oral mucosa leukoplakia. Methodological manual for doctors. М.: GOU VUNMTs, 2004. 46 р. (In Russ.)

18. Bocharova O.A., Matveev V.B., Karpova R.V. et al. Correction of immunological and clinical parameters in men with benign prostatic hyperplasia by phytoadaptogen. BEhBiM = Bulletin of Experimental Biology and Medicine 2006;141(5): 555–59 (In Russ.)

19. Chulkova S.V., Bocharova O.A., Klimenkov A.A. et al. Possibilities of increasing the effectiveness of integrated treatment of advanced gastric cancer by phytoadaptogen. Rossiysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2006;5(2):85–92 (In Russ.)

20. Khaindrava B.G., Kozina E.A., Kudrin V.S. et al. Experimental modeling of clinical and preclinical stages of Par kinson’s disease. BEhBiM = Bulletin of Experimental Biology and Medicine 2010;150(11):495–98. (In Russ.)

21. Sharp J., Riches A., Littlewood V., Thomas D. The incidence, pathology and transplantation of hepatomas in CBA mice. J Pathol 1976;119(4): 211–20. DOI: 10.1002/path.1711190405. PMID: 182943.

22. Modyanova E.A., Bocharova O.A., Ushakov V.F. The mechanical properties of hepatocytes cell-cell contacts in mice inbred strains and susceptibility to spontaneous hepatomas. BEhBiM = Bulletin of Experimental Biology and Medicine 1980;89(4):459–62 (In Russ.)

23. Bocharova O.A., Karpova R.V., Ilyenko V.A. et al. Leukocyte integrins at hepato carcinogenesis in high-cancer CBA strain of mice. Rossiysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2013;12(3):53–6 (In Russ.)

24. Bocharova O.A., Bocharov E.V., Karpova R.V. et al. Reduced occurrence of hepatomas in phytoadaptogen exposed high-cancer CBA strain of mice. Rossiysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2014;13(2):73–6 (In Russ.)

25. Bocharov E.V., Karpova R.V., Vershinskaya A.A. et al. Hepatocarcinomas infiltration lymphocytes in highcancer CBA mice when multiphytoadaptogen exposed in early postnatal ontogenesis. Rossiysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2015;14(2):85–90 (In Russ.)

26. Bocharov E.V., Karpova R.V., Bocharova O.A. et al. Multiphytoadaptogen effect in early postnatal ontogenesis improves survival and mice somatic condition of high-cancer CBA strain of mice. Rossiysky bioterapevtichesky zhurnal = Russian Journal of Biotherapy 2017;16(1):76–81 (In Russ.)

27. Deans D., Tan B., Ross J. et al. Cancer cachexia is associated with the IL10– 1082 gene promoter polimorphism in patients with gastroesophageal malignancy. Am J Clin Nutr 2009;89(4):1164–72. DOI: 10.3945/ajcn.2008.27025. PMID: 19244371.

28. Kim D., Oh S., Kwon H. et al. Clinical significance of preoperative serum interleukin-6 and C-reactive protein level in operable gastric cancer. BMC Cancer 2009;20(9):155–61. DOI: 10.1186/1471-2407-9-155. PMID: 19457231.

29. Spizzo G., Went P., Dirnhofer S. et al. Overexpression of epithelial cell adhesion molecule (Ep-CAM) is an independent prognostic marker for reduced survival of patients with epithelial ovarian cancer. Gynecol Oncol 2006;103(2):483–8. PMID: 16678891.

30. Heidemann J., Maaser C., Lјgering A. et al. Expression of vascular cell adhesion molecule-1(CD 106) in normal and neoplastic human Esophageal squamous epithelium. Int J Oncol 2006;28(1): 777–85. PMID: 16327982.

31. Sepiashvili R.I., Balmasova I.P. Physiology of natural killer. Мoscow: Мeditsina, 2005. 455 p. (In Russ.)

32. Fillon M. Biomarkers and prostate cancer progression. J Natl Cancer Inst 2011;103(21):1570–71. DOI: 10.1093/jnci/djr444. PMID: 22010173.

33. Jones S., Richards P., Scheller J., Rose-John S. IL-6 trans-signalling: the in vivo consequences. J Interferon Cytokine Res 2005;25(5):241–53. PMID: 15871661.

34. Peppa D., Micco L., Javaid A. et al. Blockade of immunosupressive cytokines restores NK cell antiviral function in chronic hepatitis B virus infection. PloS Pathol 2010;16(2):101–07. DOI: 10.1371/journal.ppat.1001227. PMID: 21187913.

35. Danese S., Semeraro S., Marini M. et al. Adhesion molecules in inflammatory bowel disease: therapeutic implications for gut inflammation. Dig Liver Dis 2005;37(11):811–8. PMID: 16168725.

36. Yu M., Kissling S., Freyschmidt-Paul P. et al. Interleukin-6 cytokine family member oncostain M is a hair-follicle-expressed factor with hair growth inhibitory properties. Exp Dermatol 2008;17(1):12–9. DOI: 10.1111/j.1600-0625.2007.00643.x. PMID: 17979974.

37. Kovalovich K., Li W., DeAngelis R. et al. Interleukin-6 protects against Fas-mediated death by establishing a critical level of anti-apoptotic hepatic proteins FLIP, Bcl-2 and Bcl-xl. J Biol Chem 2001;276(28):26605–13. PMID: 11349125.

38. Narimatsu M., Maeda H., Itoh S. et al. Tissue-specific autoregulation of the STAT3 gene and its role in interleukin-6-induced survival signals in T-cell. Moll Cell Biol 2001;21(19):6615–25. PMID: 11533249.

39. Blagosklonny M., Campisi J., Sinclair D. et al. Impact papers on aging in 2009. AGING 2010;2(3):111–21. DOI: 10.18632/aging.100132. PMID: 20351400.

40. Harrison D.E., Strong R., Sharp Z.D. et al. Rapamycin fed late in life extends lifespan in genetically heterogeneous mice. Nature 2009;460(7253):392–95. DOI: 10.1038/nature08221. PMID: 19587680.

41. Miller R.A., Harrison D.E., Astle C.M. et al. Rapamycin, but not resveratrol or simvastatin, extends life span of genetically heterogeneous mice. J Gerontol A Biol Med Sci 2011;66(2): 191–201. DOI: 10.1093/gerona/glq178. PMID: 20974732.

42. Bocharov E.V., Karpova R.V., Kazeev I.V. et al. Multiphytoadaptogen study of radioprotective activity in experiments on mice. Patologicheskaya fiziologiya i ehksperimental’naya terapiya = Pathologic physiology and experimental therapy 2013;3:55–8 (In Russ.)

43. Karpova R.V., Bocharov E.V., Kazeev I.V. et al. The radioprotective multifitoadaptogen efficacy in experiments on dogs. Patologicheskaya fiziologiya i ehksperimental’naya terapiya = Pathologic physiology and experimental therapy 2013;4:51–4 (In Russ.)

44. Belitskij G.A., Kirsanov K.I., Lesovaya E.A., Yakubovskaya M.G. The mechanisms of flavonoids anticancer action. Uspekhi molekulyarnoj onkologii = Advances in molecular oncology 2014;1:56–68 (In Russ.)

45. Nekkanti S., Tokala R., Shankaraiah N. Targeting DNA minor groove by hybrid molecules as anticancer agents. Curr Med Chem 2017;24(26):2887–2907. DOI: 10.2174/0929867324666170523102730. PMID: 28545367.

46. Oliynyk S., Oh S. Actoprotective effect of ginseng: improving mental and physical performance. J Ginseng Res 2013;37(2):144–66. DOI: 10.5142/jgr.2013.37.144. PMID: 23717168.

47. Shikov A.N., Pozharitskaya O.N., Makarov V.G. Aralia elata var. mandshurica (Rupr. & Maxim.) J. Wen: An overview of pharmacological studies. Phytomedicine 2016;23(12):1409–21. DOI: 10.1016/j.phymed.2016.07.011. PMID: 27765361.

48. Amsterdam J.D., Panossian A.G. Rhodiola rosea L. as a putative botanical antidepressant. Phytomedicine 2016;23(7):770–83. DOI: 10.1016/j.phymed.2016.02.009. PMID: 27013349.

49. Lian-Wen Qi, Chong-Zhi Wang, Chun-Su Yan. Ginsenosides from American ginseng: chemical and pharmacological diversity. Phytochemistry 2011;72(8):689–99. DOI: 1016/j.phytochem.2011.02.012. PMID: 21396670.

50. Shanmugam M.K., Lee J.H., Chai E.Z. et al. Cancer prevention and therapy through the modulation of transcription factors by bioactive natural compounds. Semin Cancer Biol 2016;40–41:35–47. DOI: 10.1016/j.semcancer.2016.03.005. PMID: 27038646.

51. Wang C., He H., Dou G. et al. Ginsenoside 20(S) – Rh2 Induces Apoptosis and Differentiation of Acute Myeloid Leukemia Cells: Role of Orphan Nuclear Receptor Nur77. J Agric Food Chem 2017;65(35):7687–97. DOI: 10.1021/acs.jafc.7b02299. PMID: 28793767.

52. Xia T., Wang Y.N., Zhou C.X. et al. Ginsenoside Rh2 and Rg3 inhibit cell proliferation and induce apoptosis by increasing mitochondrial reactive oxygen species in human leukemia Jurkat cells. Mol Med Rep 2017;15(6):3591–98. DOI: 10.3892/mmr.2017.6459. PMID: 28440403.