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and HN in the blood serum of patient with MVP considering the grade of regurgitation. The related studies demonstrated the decreasing trend of the magnesium level in patients with the 1st grade of regurgitation. The reliable decrease of the magnesium level was observed in patients with the 2nd grade, its level in blood serum decreased by 1.2 times (P < 0.05) related to the parameters of almost healthy people. We have conducted Pearson's correlation analysis in order to validate the importance of MMP-2 and MMP-9 increase with TIMP-1 in angiogenesis and conjunction with the content reduction of Mg+2, GAG and HN. Thus, in patients with the 1st grade of regurgitation we could observe the certain relation between the growth of MMP-2 and MMP-9, and the drop of TIMP-1, the ions ofMg+2 (r = -0.22-0.28, p > 0.05) and the growth of GAG level and HN activity (r = +0.24-0.31, p > 0.05) as compared to the control data. Along with that, in patients with the 2nd grade of regurgitation the expression of MMP-2 and MMP-9 had the invert correlation with the activity of TIMP-1 (r = -0.83-0.82, p < 0.01), and also after the statement of relation with the drop of the ion parameters for Mg+2 (r = -0.69-0.80, p < 0.02 and p < 0.01), and the direct correlation with the parameters of GAG (r = -0.80-0.832, p < 0.01) and HN (r = -0.77-0.81, p < 0.01). The expression of TIMP-1 correlated with the drop of the level of Mg+2 ions in blood (r = -0.75, p < 0.01)
and inversely with the GAG indicators (r=-0.84, p < 0.01) and HN activity (r = -0.86, p < 0.01).
Therefore, the study of the mechanism of cardiac hemodynamics disorders showed that in the patients with MVP and regurgitation we can observe the inadequate endothelial production of antiangiogenic factors, which results in the decrease of response relaxation, and determines the drop of responsiveness index and the risk of hypertensive state, in particular induced by physical exercises. The development of these changes is conditioned by the rise of VEGF in blood serum; it determines the risk of angiogenesis stimulation in patients with MVP, the expression of which is related with the regurgitation grade. In patients with congenital MVP of the 1st and 2nd grade of regurgitation, we can diagnose endothelial dysfunction, caused by the imbalance of NO-system. The other factors that lead to the MVP formation includes the drop of magnesium level, HN activation, which promotes the increased degradation of extracellular matrix components followed by the growth of its debris excretion, this is associated with the activation of MMP-2 and HN. It can be expected that the reason for the progression of mitral regurgitation in patients with congenital MVP lies in the drop of TIMP-1 inhibiting action, the content of Mg+2 ions, HN activation.
References:
1. Kadurina T. I., Gorbunova V. N. Connective tissue dysplasia. - St. Petersburg: ELBI, 2009. - 714 p.
2. Gnusaev S. F. Connective tissue cardiac dysplasia syndrome in children//Consulting physician. - 2010. - No. 8. - P. 40-44.
3. Pereskalskaya M. A., Makarova L. I., Vereshchagina G. N. Neuroendocrine dysfunction in women with systemic connective tissue dysplasia//Clinical medicine. - 2002. - V. 80, No. 4. - P. 48-51.
4. Komissarova L. M., Karachaeva A. N., Kesova M. I. Gestation and partus course in CTD//Obstetrics and Gynec. - 2012. - No. 3. - P. 4-8.
5. Domnitskaya T. M., Diachenko A. V., Kupriyanova O. O., Domnitskiy M. V. Clinical relevance of Magnesium orotate administration in adolescents with connective tissue cardiac dysplasia syndrome//Cardiology. - 2005. - V. 45(3). - P. 76-81.
6. Kadurina T. I., Abbakumova L. N. Rehabilitation principles for patients with connective tissue dysplasia//Consulting physician. -2010. - V. 40. - P. 10-16.
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Shukurova Umida Abdurasulovna, PhD, Tashkent State Dental Institute, Uzbekistan, Faculty Dental Therapeutics department E-mail: shua1981@mail.ru Bekjanova Olga Esenovna, PhD, MD, DDS, Faculty Dental Therapeutics department E-mail: bekjanovaolga@mail.ru
Significance of mid mass molecules in cytokine cascade of oral fluid in patients with oral lichen planus
Abstract: Taking into consideration that severe disturbances in metabolism accompanying by tissue destruction occur in the organism along with accumulating of toxic metabolites in biologic environment, the study of the problem is seen as endotoxicosis. The analysis of such a correlated interconnection certifies various influence of MMM on the immune system of the organism.
This influence is complicated and specific and is reflected in immune imbalance response and obviously in the structural disturbances of oral mucous membranes epithelium.
Keywords: oral lichen planus (OLP), pro- inflammatory cytokines, mid mass molecules (MMM), nuclear derivative, toxic derivative, aromatic derivative.
Oral lichen planus (OLP) is one of the most common diseases of mucous membrane of the oral cavity (MMOC). In the structure of MMOC diseases OLP composes 35 %. It is known that in OLP of MMOC main changes take place in the epithelium of mucous membranes [1, 156-166; 10, 13-14].
At present, main metabolic chains responsible for cellular damage and damage of tissues in the pathology have been noted. They are: high level of peroxide oxidation of lipids and the lack antioxidant defense, unbalanced cytokine level with prevailed level of pro- inflammatory cytokines [1, 160-162; 4, 14-16; 8, 89-90; 9, 134-135; 10, 11-12].
Section 4. Medical science
Taking into consideration that severe disturbances in metabolism accompanying by tissue destruction occur in the organism along with accumulating of toxic metabolites in biologic environment, the study of the problem is seen as endotoxicosis. Being universal, the syndrome of endogenous intoxication (EI) is able to influence negatively on the course of various pathologic processes in the organism [3, 113-114; 5, 144-145; 6, 140-141].
Investigations of EI intensity in interaction with oral fluid cytokines in patients with various clinic forms of OLP MMOC have not been done.
Purpose of the investigation is the analysis of mid mass molecules (MMM) changes of oral fluid cytokines as well as evaluation of correlation between these data.
Materials and methods. 139 patients with OLP of MMOC including 55 males (39.57 %), 84 females (60.43 %) have been examined. 20 people of comparing gender and age composed a control group. In 36 patients (25.9 %) a typical form of the disease was diagnosed; exudative hyperemic form was in 34 (24.4 %); erosive-ulcerous form was in 42 (30.22 %); bullous form was diagnosed in 27 patients (19.42 %).
In verification of the disease forms, Borovskiy and Mashkil-leison's classification was used (2001). Comparing groups were randomized according to their gender and age. This provided representation of received data.
Oral fluid was taken in the morning on empty stomach before drug therapy, it was centrifuged at 1500 cir/min, during 20 minutes.
Cytokine profile was studied by immuno-enzymes analysis using "Cytokine" Ltd. device, St. Petersburg. Evaluation of endogenous intoxication in oral fluid was performed in MMM spectrum by screening [7, 72-73].
Three Derivatives of MMM have been noted:
1. Nuclear derivative — is determined at 230 nm. wave represented by hystone proteins and DNA wastes.
2. Toxic derivative — 254 nm. of wave consisting of hydrophobic toxins with high biologic similarity structures, contains products of not complete disintegration of proteins.
3. Aromatic derivative — 280 nm. of wave, containing aromatic aminoacids (mediators, hormones).
Table 1. - MMM spectrum in
Results were shown by the symbolic units. Aromatic index (AI) was calculated (E280/254 nm.) as well as nuclear-peptide index — (NPI = E230/E254 nm).
Connections ofvarious signs were estimated by linear correlation. Results and discussion
In patients with OLP ofMMOC oral fluid has some increase in the data of endogenous intoxication.
Growth of clinical symptoms of OLP was associated with significant increase of level in toxic derivative - E-254 nm.; nuclear derivative - E-230 nm.; increase in nuclear — peptide index, decrease in aroma index (AI).
Thus, the value toxic E-254 nm. derivative were much higher than in control group up to 69.69 % (P < 0.01) peptide — nuclear — to 208.56 % (P < 0.01), nuclear — peptide index - to 77.42 % (P < 0.01), aroma index was decreased to 37.78 % (P < 0.01). Together with growing of the severity of the process on mucous membranes of the oral cavity increase in EI is noted. Quantity of toxic E-254 nm., derivative in typical type of OLP was 27.58 % (P < 0.01) higher than in control group; exudative-hyperemeric - 46.36 % (P < 0.01); erosive-ulcerous and bullous — 112.12 % (P < 0.01) and 88.18 % (P < 0.01) correspondingly; the dynamics of nuclear derivative E-230 nm. shows 46.39 % (P < 0.01); 115.46 % (P < 0.01); 376.29 % (P < 0.01) and 293.81 % (P < 0.01) nuclear-peptide index 18.39 % (P < 0.01); 54.84 % (P < 0.01); 112.90 % (P < 0.01) and 96.77 % (P < 0.01) correspondingly decrease of AI is 19.78 % (P < 0.01); 32.97 % (P<0.01) 50.55 % (P < 0.01) and 45.46 % (p < 0.01) (Table 1).
Significant peculiarity of MMM is in their clearly marked high biologic activity. Accumulation of MMM is not only a marker of endotoxication, they also contribute to severances of the pathologic process acting as secondary toxic agents influencing all vital system and organs. Data of the MMM level is the main biologic marker reflecting the level of the pathologic protein metabolism [5, 141-142].
As the main function in metabolism changes is referred to proteins, directly or indirectly, the increase of MMM level mostly due to peptide nature is one of the general pathobiochemical mechanisms of epithelial barrier and progression of erosive- ulcerous damage in the oral cavity (Table 1). oral fluid of the patients with OLP
Clinical forms of OLP Datum in optic density Indices
Е 280 Е 254 Е 230 NPI IA
Control 0.302 ± 0.011 0.330 ± 0.014 0.097 ± 0.004° 0.31 ± 0.012° 0.91 ± 0.03°
Typical 0.306 ± 0.009 0.421 ± 0.02 • 0.142 ± 0.005 • 0.46 ± 0.02^ 0.73 ± 0.02^
Exudative — hyperemic 0.297 ± 0.011 0.483 ± 0.015^'° 0.209 ± 0.001 0.48 ± 0.015 ^ 0.61 ± 0.02 ^
Erosive- ulcerous 0.320 ± 0.014 0.700 ± 0.028 •'°'*'A 0.462 ± 0.02 •>°>*>A 0.66 ± 0.031 •'°'*'A 0.45 ± 0.01 •'°'*'A
Bullous 0.311 ± 0.012 0.621 ± 0.031 •'"'* 0.382 ± 0.012 0.61 ± 0.029^'°'* 0.50 ± 0.02
On average in patients with OLP 0.308 ± 0.008 0.560 ± 0.025 • 0.299 ± 0.008^ 0.55 ± 0.016^ 0.57 ± 0.01^
Note: • — P < 0.05 for the control group; ° — P < 0.05 for the typical form; * — P < 0.05 for the exudative-hyperemic form; A — P < 0.05 for bullous form.
Table 2. - Characteristics of cytokines status of oral fluid according to clinical form of OLP
Cytokine pg/ml Typical Exudative- hyperemic Erosive-ulcerous Bullous OLP-in general Control group
FNO-a 70.81 ± 2.35^ 100.02 ± 5.0T'° 650.0 ± 55.40^ 550.26 ± 31.24 ^ 358.25 ± 10.11 25.33 ± 1.05
IFN-Y 58.32 ± 1.65 • 70.25 ± 3.24^° 424.02 ± 70.0^ 325.43 ± 25.42 ^ 241.25 ± 11.08 24.32 ± 0.92
IL-1 22.63 ± 0.95 • 60.81 ± 2.48^° 562.08 ± 44.31 • 462.08 ± 24.81 ^ 142.08 ± 7.02 10.21 ± 0.58
IL-6 30.82 ± 1.44 • 71.25 ± 3.43^° 505.30 ± 30.22^ 421.0 ± 30.77^°'* 181.32 ± 8.61 14.81 ± 0.69
IL-8 26.81 ± 1.22 • 64.83 ± 2.74^'° 425.41 ± 40.25 • 324.32 ± 28.42 ^ 224.31 ± 7.82 13.62 ± 0.55
IL-4 30.33 ± 1.24 • 22.62 ± 1.02^° 9.32 ± 0.25 • 10.32 ± 0.48 ^ 17.65 ± 0.55 16.05 ± 0.71
IL-10 34.32 ± 1.62 • 27.84 ± 1.21^° 8.31 ± 0.3^ 9.11 ± 0.31 ^ 19.40 ± 0.62 18.22 ± 0.24
Note: " — P<0.05 for the control group; ° — P<0.05 for the typical form of OLP; * — P<0.05 for the exudative- hyperemic form; A — P<0.05 for bullous form of OLP.
So, in patients with OLP monodirected increase of MMM content in oral fluid was noted mostly in patients with erosive-ulcerous form.
Evaluation of MMM content in oral fluid enables to assess patients condition individually as well as the severity of the course of OLP, the ways of pathogenetic treatment in the case of mucous membranes of the oral cavity.
Deviations in cytokine profile in the oral fluids were manifested by imbalance of pro- inflammatory mediators. At the same time in favorable cases (typical and exudative — hyperemic) the increase in anti- inflammatory mediators activity was noted: IL-4 and IL-10 correspondingly to 88.97 % (P < 0.01) — 40.99 % (P < 0.01) and 88.36 % (P < 0.01) — 51.66 % (P < 0.01); in severe forms of erosive-ulcerous and bullous type some decrease to 35.70 % (P < 0.01) — 41.93 % (P < 0.01) and 54.4 % (P < 0.01) — 50.0 % (P < 0.01) correspondingly.
In all clinical forms of OLP the content of pro- inflammatory cytokine is clearly (P < 0.05) higher than the values of control groups and progressively was growing with the severity of the process in mucous membranes. So, in typical form the index FNO-a raised up to 179.55 % (P < 0.01) in erosive-ulcerous form - to 2089.77 % (P < 0.01) corresponding dynamics of IFN-y composed 139.80 % (P < 0.01) — 1643.5 % (P < 0.01); IL-1 to 121.65 % (P < 0.01) — 5405.20 % (P < 0.01); Il-6-to 108.10 % (P < 0.01) — 3314.12 % (P < 0.01) and IL-8 to 96.84 % (P < 0.01) — 3023.42 % (P < 0.001) (table 2).
Thus, as the erosive- ulcerous damage of the mucous membrane develops moderate activation of anti- inflammatory factors is changed into their inhibition on the background of intensively growing pro- inflammatory mediators (by ten, thousand times).
When correlating the data the following dependence between MMM content and cytokine level were noted:
• In patients with typical form of the disease coefficient unit of correlation certify moderate positive connection between the data in the investigation: coefficient unit range between 0.4-0.62.
• In patients with exudative-hyperemic form along with the growing correlative interconnections between pro- inflammatory cytokines and MMM level, coefficient unit is
noted to be at 0.64-0.72; and interconnections with antiinflammatory cytokines are reduced up to 0.32-0.36.
• In patients with erosive-ulcerous and bullous forms interconnections between the pro-inflammatory cytokines level MMM concentration rises up to 0.77-0.88; but the direction of interconnections with anti- inflammatory cytokines becomes negative and is noted as moderate (-0.52) and high (-0.76).
It can be supposed that mucous membrane in no sign course of typical form OLP has a balance between immune cells produced by pro-and anti- inflammatory cytokines and endotoxins.
Under the influence of stress factors, along with developing endotoxicosis in pathogenic concentrations earlier primitive cells producing cytokines are activated. The character of further series of reaction depends on general balance of produced pro- and antiinflammatory cytokines.
In the case of balance disturbance, invasion of extra MMM into the mucous membranes macrophages are activated and secrete mediators to initiate lymphocytes for cytokine production.
Thus, in mucous membranes in the areas of papulous efflorescence are formed which are sensitive to MMM effect and pro- inflammatory cytokines (FNO-a, IFN-y) [8,155-156; 11, 364-365].
In critical concentration of MMM and pro- inflammatory cytokines significant disturbances of local and systemic metabolism appear as well as immunologic disturbances, favorable conditions for the formation of inflammation focus.
Together with progress of the process in the mucous membranes and growth of endogenous intoxication and strengthening of systemic reaction of the organism to the local process we can see the development of imbalance between endotoxic aggression and cytokines negative processes are progressive.
Endless cytokine production leads to aggression of mediators and further progressive systemic inflammatory reactions, severity of the local damage of the mucous membranes.
The analysis of such a correlated interconnection certifies various influence of MMM on the immune system of the organism.
This influence is complicated and specific and is reflected in immune imbalance response and obviously in the structural disturbances of oral mucous membranes epithelium.
References:
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