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Section 5. Medical science
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12. Nazirov F. G., Khudaybergenov Sh. N., Eshonkhodjaev O. D. The method.recommendations. - Tashkent, 2016.
Bakhritdinova Fazilat Arifovna, Tashkent Medical Academy, Department of eye diseases, Doctor of medicine, professor E-mail: bakhritdinova@mail.ru
Narzikulova Kumri Islamovna, Department of eye diseases, PhD, senior teacher E-mail: kumri78@mail.ru
Mirrakhimova Saidakhon Shukhratovna, Department of eye diseases, PhD, docent E-mail: saidakhon.m@mail.ru
Khera Akshey,
ophthalmologist, Ophthalmological Clinic "Vedanta", Tashkent
E-mail: retinauz@gmail.com
Biochemical parameters of the effect of laser radiation in the experiment
Abstract: Was studied effect of different modes of photodynamic irradiation in 80 rats on indicators of endogenous intoxication in dynamics.
PDT in 300 mJ pulsed mode (630 nm. for 3 minutes) does not cause endogenous intoxication and intensification of lipids peroxidation (LPO), that allow to use the appropriate mode in the practice of ophthalmology. Using of 300 mJ pulse mode (890 nm. for 3 minutes), 60 J PD mode (630 nm, 15 seconds) and particularly 120 J PD mode (630 nm. for 30 seconds) leads to gradual accumulation of average-weight molecules (AWM) and intensification of LPO in the circulating blood of animals, that leads to development of endogenous intoxication, therefore such dosage cannot be used in vision organ.
Keywords: photodynamic therapy, photodynamic doses, experiment, endogenic intoxication, molecules of average molecular weight, malon dialdegidis.
Introduction. Achievements of photobiology laid a solid foundation for the development and practical application of lasers, including the field of ophthalmology. There are widely known stimulatory, microcirculatory and anti-inflammatory effects of therapeutic doses of radiation exposure. It is appropriate to mention that one of the promising areas in ophthalmology is photodynamic therapy based on the selective destruction of proliferating cells as a result of photochemical reactions caused by reacting of accumulating therein photosensitizer (PS) which is activated by the light of particular wavelength [4, 25-30; 9, 55-58; 10, 167; 11, 155-162].
The development and introduction of photodynamic therapy (PDT) and fluorescence diagnosis (PD) in the treatment of cancer has caused deep interest ofresearchers to the possibility of using these techniques in other fields of medicine and in particular in ophthalmology [7, 39-40; 8, 23-25;13, 45-51; 15, 740-746; 18, 228-232].
Today, the perspectives of PDT treatment are associated with treatment of choroidal neovascularization of various etiologies, especially in age-related macular degeneration, diabetic retinopathy, high myopia complicated and ocular oncologic diseases [2, 45-47; 3, 265; 12, 66-68; 14, 752-763; 15, 740-746; 17, 747-752].
Over the last 5 years, there are many publications concerning the use of PDT at non-cancer ophthalmic diseases, however,
it should be noted that very few studies are devoted to the fundamental aspects of this trend [13, 45-51]. At the same time, such an important aspect as the impact of the so-called photodynamic doses (PDD) on biological tissues is not studied yet.
Having a cytotoxic effect PDT undoubtedly affects the processes of oxidative stress in the body. However, there is no information in the available literature on the study of the effects on the processes of lipid peroxidation (LPO), depending on the mode of action of PDT [5, 3-8; 6, 15-18].
The introduction of PDT in clinical practice of ophthalmology in Uzbekistan was significantly constrained by high cost of foreign trade photosensitizers (PS) and laser equipment.
In Uzbekistan there was developed an apparatus working in a range of630 nm., output power up to 5 W. (Mavlyan-Khodzhaev R. Sh. and S. Sadykov R. A., 2009) and was used in purulent infected hardhealed wounds in surgery, dentistry and dermatology [9, 55-58; 10, 167].
However, there are still no works on the application of PDT in the treatment ofeye diseases in Uzbekistan, using domestic laser systems.
Objective: To evaluate the effect of electromagnetic radiation on the eyes of rats at doses used for photodynamic therapy, on the level of malon dialdegidis (MDA) and its relationship with indicators of endogenous intoxication in rats in the experiment.
Biochemical parameters of the effect of laser radiation in the experiment
Material and methods
Research was conducted on 80 white mongrel rats. A burn of cornea had been performed to all the animals. Laser radiation was performed to the projection of the right eye (5 mm. aperture) daily by 7 irradiations per course, on the domestic ALT "East" equipment.
Depending on the dose of exposure the animals were divided into 5 groups of10 animals each — control (intact animals) and 4 experimental groups:
1) 300 mJ pulse mode (control group — 890 nm, 3 min.);
2) 300 mJ pulse mode (630 nm. for 3 minutes);
3) 60 J PDD (630 nm. for 15 seconds);
4) 120 J PDD (630 nm. for 30 seconds).
Animals were sacrificed after 3, 7 and 14 days of application of PDT. All the experiments on animals were conducted in accordance with the recommendations of WHO's work with experimental animals, and with caution. An euthanasia was performed to rats when their condition was assessed as moribund.
Determining the level of the average molecular weight (AMW) in the serum was performed by the method of I. M. Korochkin et al. (1984) [5, 3-8]. To 0.2 ml. of the bioassay there was added 0.2 ml. of 16 % solution of trichloroacetic acid (TCA) and the proteins were precipitated by centrifugation. 2.7 ml. of distilled water was added to 0.3 ml of the supernatant and measured for UV absorption with a spectrophotometer at a wavelength of254 nm. in quartz cuvettes with a pathlength of 1 cm. versus blank sample. In the blank sample 0.1 ml. of the TCA solution was added to 2.9 ml. of water. The level of AMW was expressed in units.
Table 1. - Content of MDA in the blood
Determining the level ofmalon dialdehyde (MDA) in the serum was performed by the method ofA. I. Andreeva et al. [1, 41-43].
Results and discussion
Studies have shown that in animals of the 1st group treated with 300 mJ pulse mode (890 nm. for 3 minutes), the level of MDA in serum increased to 3.17 times on the third day ofexposure (Table. 1). Intensification of lipid peroxidation stayed further, only to the end of the experiment we observed a decrease in 2 times in comparison with the values ofprevious periods, but they significantly exceeded the indicators of intact rats in 1.5 times. The third group of animals irradiated with 60 J in PD mode (630 nm.) for 15 sec. however the content of MDA increased significantly early after exposure but in future we observed a significant increase of it in 1.47 times. Using a more powerful mode of exposure (Group 4) resulted in an intensification oflipid peroxidation: significant increase of the MDA level in 2.05; 1.97 and 1.98 times, respectively timing after 3, 7 and 14 days after irradiation.
Analyzing the results of endogenous intoxication, evaluated by the level of AWM ther should be noted the coincidence of the dynamics of its changes to the index of hyperlipoperoxidation. Thus, in the animals of the Group 1 the level ofAWM in serum has increased sharply to 2.37 times on the 3rd day, further it gradually reduced, but still left higher than the values in intact rats (Table. 2).
In the 2nd group of indicators of endotoxemia were within the reference values. The third group of animals' indicators AWM in blood serum on the 3rd and 7th day were within normal values, but in the future we saw their increase. Using a more powerful mode of exposure (Group 4) resulted in a gradual accumulation of AWM in the circulating blood of animals. serum of experimental animals, M ± m
Groups Terms of the study, days
3 7 14
intact 5.12 ± 0.37
1) 300 mJ pulse mode (890 nm. 3 min.) 16.24 ± 1.22*** 15.38 ± 1.08*** 7.69 ± 0.45**
2) 300 mJ pulse mode (630 nm. 3 min.) 6.55 ± 0.47** 5.13 ± 0.33 5.15 ± 0.28
3) 60 J PD mode (630 nm. 15 sec.) 6.12 ± 0.29** 5.59 ± 0.38 7.55 ± 0.33*
4) 120 J PD mode (630 nm. 30 sec.) 10.05 ± 0.54*** 10.09 ± 0.41*** 10.12 ± 0.32***
Groups Terms of study, days
3 7 14
Intact 0.091 ± 0.007
1) 300 mJ pulse mode (890 nm. 3 min.) 0.210 ± 0.012 0.187 ± 0.013 0.165 ± 0.009*
2) 300 mJ pulse mode (630 nm. 3 min.) 0.110 ± 0.008 0.102 ± 0.008 0.083 ± 0.007
3) 60 J PD mode (630 nm. 15 sec.) 0.092 ± 0.007 0.094 ± 0.008 0.116 ± 0.011*
4) 120 J PD mode (630 nm. 30 sec.) 0.122 ± 0.009 0.137 ± 0.010* 0.187 ± 0.012*
Note: * — the difference between the indices of intact and experimental groups reliable, P < 0.05 Therefore, when using the pulse mode of PDT 300 mJ (630 nm. for 3 minutes) the intensification of peroxidation and the development of endogenous intoxication are not observed. Our results correspond to some extent to the results obtained by R. A. Sady-kov et al. (2012), which have shown the reduction ofleukocytes intoxication index (LII) from 6-8 conventional units to 1.95 conventional units in patients with purulent wounds after a course of PDT with MB after 3 days, to the 10th days of comprehensive treatment of the value of FRI meet regulatory [6, 15-18]. According to the authors, this condition is associated with the increase of nonspecific resistance of patients' organisms and the reduction of intoxication. There was also marked a decrease in terms of clearance of the wounds from the detritus and purulent fibrinous masses, appearance
of granulation and beginning of epithelialization. One of the mechanisms of such positive action is a maximum antibacterial effect against multi-drug resistant strains of microorganisms.
Based on these data, the following conclusions:
1. Application of PDT in 300 mJ pulse mode (630 nm. for 3 minutes) does not cause endogenous intoxication and intensification of lipid peroxidation.
2. PDT with high power leads to an intensification of lipid peroxidation, the severity of which depends on the duration of exposure. In the 300 mJ pulse mode (890 nm. for 3 minutes), 60 J PD mode (630 nm., 15 seconds) and 120 J PD mode for 30 minutes there observed the gradual accumulation of lipid peroxidation products and development of endogenous intoxication.
Note: * — accuracy of the relative indicators of intact rats (* — p < 0.01; ** — p < 0.05; *** — P < 0.001).
Table 2. - Content of AWM in the blood serum of experimental animals, according to doses of PDT, n = 6-7, M ± m
Section Б. Medical science
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Bakhritdinova Fazilat Arifovna, Tashkent Medical Academy, Department of eye diseases, Doctor of medicine, professor E-mail: bakhritdinova@mail.ru Mirrakhimova Saidakhon Shukhratovna, Department of eye diseases, PhD, docent E-mail: saidakhon.m@mail.ru Karimov Ulugbek Rasulovich, Sirdarya Regional Ophthalmic Hospital, Departamant of microsurgery, Ms.D. E-mail: dr_karimov@mail.ru Narzikulova Kumri Islamovna, Department of eye diseases, PhD, senior teacher E-mail: kumri78@mail.ru
The results of medicated decreasing of intraocular pressure at neovascular glaucoma
Abstract: The usage of combined fixed, hypotensive drug brimoptic in NVG leads to an authentic decrease IOP, especially in NVG second stage (p < 0.05). In third stage of NVG, given drug is recommended for decrease of IOP in preoperative
period of fistulaized operation.
