The role of perlecan in remodeling the extracellular matrix of the liver, lungs and spleen of mice after administration of BCG vaccine and the liposome-encapsulated dextrazide

Abstract

Despite significant advances in the diagnosis of tuberculosis, the issues of the mechanism of development and treatment of fibrotic manifestations are relevant and take place in the treatment. The aim of the study was to study the content of perlecane and to find out its participation in the remodeling of the extracellular matrix of the organs of mice with tuberculous inflammation and when they were injected with an antitubercular composition. Materials and methods. The experiment was carried out on male mice of the BALB/c line, which were divided into 4 groups of 5 individuals each. All animals, with the exception of intact mice (group 1), were retroorbitally injected with BCG vaccine once (0.5 mg of microbial bodies in 0.2 ml of 0.9% NaCl solution) to simulate a generalized tuberculosis process. After infection 6 mo for 3 mo, group 2 mice were intraperitoneally injected with 50 µl of 0.9% NaCl solution, group 3 - intraperitoneally with 50 µl of liposome-encapsulated dextrazide solution (LEDZ, isoniazid conjugate and oxidized dextran enclosed in liposomes), group 4 - LEDZ solution was inhaled by spraying in the chamber through a nebulizer for 5 min at the rate of 50 µl of solution per animal. After the last administration of the NaCl and LEDZ solution, mice were removed from the experiment, organs were taken, and tissue homogenates were prepared. The content of perlecane, hyaluronan, protein-bound hydroxyproline, tissue metalloproteinase inhibitors (TIMP-1, TIMP-2) and the activity of matrix metalloproteinases (MMP), hyaluronidases, α2-macroglobulin were measured. Results. In all organs of infected mice, the content of perlecan, hyaluronan, protein-bound hydroxyproline, TIMP-1, TIMP-2 was increased, MMP, hyaluronidase, α2-macroglobulin were activated. Regardless of the method of administration of LEDZ, the severity of fibrosis decreased due to a decrease in the content of hyaluronan in all organs and protein-bound hydroxyproline in the lungs. Perlecan with intraperitoneal administration is involved in fibrosis of the spleen, inhalation - reflects the severity of liver fibrosis. Conclusion. Correlations of perlecan with extracellular matrix components reflect participation in the regulation of fibrosis of organs in BCG-induced inflammation.

Mechanism of lungs fibrosis in mycobacterial infection

“…Under the intraperitoneal LEDZ administration in the stage of chronic BCG granulomatosis (6 months after infection with BCG vaccine, LEDZ administration for 3 months), the lung contents of hyaluronan, perlecan, peptide-bound hydroxyproline, and protein-bound hydroxyproline decreased, but the content of free hydroxyproline increased (Figure 1D), indicating increased degradation of collagens and suppression of their synthesis [123,130]. The introduction of LEDZ at this stage of BCG granulomatosis led to a decrease in the activity of MMPs and specific and non-specific inhibitors (TIMP-1, TIMP-2, and α2-MG), which also implies a decrease in the activity of destructive-inflammatory potential [122]. The combined decrease in MMPs and α2-MG activity, level in TIMP-1 and TIMP-2 indicates the suppression of protease, antiprotease activity, and inflammatory response due to the influence of LEDZ.…”

“…According to the authors [121], glycolipid lipoarabinomannan and other Mtb components induce the expression of MMP-9. In the mouse model of BCG-induced inflammation, 6 months after infection, the total activity of MMPs increased by 9 times, hyaluronidase by 4 times, α2-MG by 10 times, TIMP-1 by 11 times, and TIMP-2 by 13 times in the lungs [122,123], that indicates high both protease and antiprotease activity of enzymes.…”