RU2456678C1 - Method for preventing and treating hepatic consequences of ischemia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: liver is resected in an animal experimentally. A pharmacological protection is presented by Mexicor and Serotonine Adipinate. On the operation day, 5% Mexicor is used in the form of intravenous slow stream introductions twice - 35 minutes before the operation in a dose of 5 mg/kg of weight and 11 hours after the first introduction in a dose of 7 mg/kg of weight. Serotonine Adipinate dissolved in 5 ml of isotonic solution NaCl is used in the form of intravenous slow stream introductions three times - 1 hour before the operation in a dose of 0.15 mg/kg of weight, 2 hours after the first introduction in a dose of 0.2 mg/kg of weight, 10 hours after the first introduction in a dose of 0.1 mg/kg of weight. During the postoperative period, Mexicor and Serotonine Adipinate are introduced for 8 days. Mexicor is introduced three times a day at 9 o'clock in a dose of 2.5 mg/kg of weight, at 15 o'clock in a dose of 1.5 mg/kg of weight, at 21 o'clock in a dose of 6 mg/kg of weight. Serotonine Adipinate is introduced twice a day - at 8 o'clock in a dose of 0.15 mg/kg of weight, at 17 o'clock in a dose of 0.1 mg/kg of weight.

EFFECT: method provides prolonging a period of safe hepatic exsanguination, ensures treating and preventing ischemic and reperfusion injuries of hepatic tissues and microcirculatory bloodstream, promotes preventing major bleedings accompanying hepatic operations.

3 ex, 3 tbl, 3 dwg

Description

The invention relates to medicine, namely to experimental surgery, and can be used when performing a liver resection.

With liver resections, the problem of intraoperative bleeding with massive blood loss is one of the most important (Gantsev Sh. Kh. Intraoperative perfusion of the liver during surgical interventions on the abdominal organs. // Soviet medicine. - 1987. - No. 10. - P.10-12). The most effective and optimal way to prevent this is to squeeze the hepatoduodenal ligament, which allows the surgeon to perform temporary hemostasis and to carry out important stages of the operation with minimal blood loss (Veronsky G.I. Anatomical and physiological aspects of liver resection. - Novosibirsk: Nauka, 1983. - 185 from). But at the same time, portal hypertension, hypoxia and liver ischemia develop, hemodynamics are destabilized, which, when the 20-minute threshold is exceeded, can lead to irreversible morphological and functional changes in the liver, up to necrotic changes in hepatocytes (Antopolskaya E.V. Correction of ischemic liver damage during resection of the liver bleeding conditions: Abstract of thesis ... Candidate of Medical Sciences. - Kharkov, 1988. - 21 p.).

To prevent and correct metabolic disorders in the cell, various drugs with an antioxidant effect are used - dibunol, sodium oxybutyrate, cytochrome C, etc. (Bilenko M.V. Theoretical and experimental justifications for the use of antioxidant therapy for the prevention of acute ischemic damage in organs. : Bioantioxidants in the regulation of metabolism in normal and pathological conditions. - M., 1982. - P.195-213; Okovity S.V. Clinical pharmacology of antihypoxants (part II). // PHARMindeks-Practik. - 2004, 7 - P.48 -63), but existent uyuschy their arsenal is limited, and many well-known application techniques can not always prevent the occurrence of and to stop already developed ischemic lesion of liver parenchyma.

Another direction in the fight against hepatic ischemia is the desire to protect the microvasculature of the liver through the use of drugs with different mechanisms of action. In particular, attempts are being made to influence the development of smooth muscle insufficiency of microvessels (A. P. Simonenkov. V. D. Fedorov, V. M. Klyuzhev, V. N. Ardashev. The use of adipinate serotonin to restore impaired smooth muscle function in surgical and therapeutic patients. // Bulletin of intensive care - 2005 - No. 1. - P.1-6; Simonenkov AP, Fedorov VD On the unity of tissue hypoxia and shock. // Anest. and resuscitation. - 2000 - No. 6 . - P.73-76; Simonenkov A.P., Fedorov V.D., Fedorov A.V. et al. Mechanism of endogenous vasomotrica and smoothoma echnoy failure microvasculature // Bulletin of Medical Sciences -.. 1994. - №6 -. S.11-15). However, the applied methods do not yet allow to significantly increase the duration of liver anoxia.

The closest is a method for correcting ischemic damage to the liver under conditions of bloodlessness, the essence of which is intravenous administration to an experimental animal (dog) of 5 mg of 1% solution of serotonin-adipinate before and within 30 minutes after compressing the hepatoduodenal ligament in order to prevent the development of smooth muscle insufficiency of the microvasculature of the liver (patent No. 2134576 from 03/11/1998) - analogue, it is also a prototype. This method helps to eliminate smooth muscle insufficiency of the microvasculature, thereby improving endogenous vasomotor.

The disadvantage of this method is that the authors limited themselves to the use of a drug that regulates microcirculation, not trying to correct other inevitably occurring disorders caused by exposure to hypoxia, and a twofold administration of serotonin adipate is still not enough for the treatment and prevention of hypoxic and reperfusion injuries of the liver tissue and its microvasculature .

There is also known a method for the correction of ischemic liver damage (patent No. 2394574 from 05/20/2009), which, against the background of the application of a temporary extracorporeal portocaval shunt under conditions of bleeding of the organ, involves intravenous administration of actovegin and Mexicor. In addition, each of the drugs, both before and after the operation, is administered once a day. The use of this method helps to improve metabolic processes and reduce the severity of damage to the hepatic parenchyma.

The disadvantage of this method is the imposition of an extracorporeal portocaval shunt, when used with small diameters it does not bear any payload, and with increasing diameters, the development of hepatic toxemia (including azotemia) cannot be ruled out, which can lead to toxic effects on many organs and systems (D.E. Kutepov, V.N. Semenov, A.Yu. Denisov, I.N. Pasechnik. Use of extracorporeal treatment methods in the treatment of liver failure. // Bulletin of intensive care. - 2004. - No. 2. - S.65-70; Aron son L., Gacad R.C., Kaminsky-Russ K. et al. Evidence of gut production of endogenous benzodiazepines: implications for hepatic encephalopathy. // Gastroenterology. - 1996. - Vol. 110. - P.1144). The disadvantages of this method also include the introduction of each of the drugs once a day, which, taking into account their pharmacodynamics and pharmacokinetics, from our point of view, is not enough.

In both methods considered, the duration of treatment with the drugs used is insufficient, since the duration of the drug exposure should take into account possible or occurring morphological and biochemical changes in the liver.

The mechanisms of development of ischemic damage to the liver are complex, therefore, the impact on any one link in the formation of the pathological chain is not enough. Considering the above, monotherapy for hypoxic conditions seems to us insufficient, we consider the use of complex methods for the correction of liver ischemia to be relevant and promising.

The objective of the invention is the treatment and prevention of ischemic and reperfusion injuries of the liver tissue and its microvasculature arising from resection of the liver in the conditions of its temporary shutdown from the blood circulation, prolonging the safe period of bleeding of the liver in order to prevent massive bleeding during operations on this organ.

The technical result is that the animal (dog) on the day of surgery, Mexico in the form of a 5% solution is intravenously slowly injected twice - 35 minutes before surgery at a dose of 5 mg / kg body weight, 11 hours after the first injection at a dose of 7 mg / kg body weight, serotonin adipate, diluted in 5 ml of isotonic NaCl solution, is injected intravenously slowly three times - 1 hour before surgery at a dose of 0.15 mg / kg body weight, 2 hours after the first administration at a dose of 0.2 mg / kg of weight, 10 hours after the first injection at a dose of 0.1 mg / kg of weight; in the postoperative period, each of the drugs is administered for 8 days: Mexico - three times a day - at 9 ⁰⁰ at a dose of 2.5 mg / kg of body weight, at 15 ⁰⁰ at a dose of 1.5 mg / kg of body weight, at 21 ⁰⁰ at a dose of 6 mg / kg body weight, serotonin adipate - twice a day - at 8 ³⁰ in a dose of 0.15 mg / kg body weight, at 17 ⁰⁰ in a dose of 0.1 mg / kg body weight.

The pharmacological effect of serotonin is due to the fact that it contributes to the normalization of the smooth muscle function of blood vessels (endogenous vasomotoria), reduces local hypoxia of internal organs, improves metabolism in areas of hypoxia. It is well absorbed and distributed to organs and tissues, remaining in the blood plasma for 8 hours (Russian Medicines Register of the Radar Station 2008. - M., "Radar Station 2008", 2007 - P.800-801).

The pharmacological effect of Mexico is determined by the inhibition of lipid peroxidation, increased activity of the antioxidant system, activation of the energy synthesizing functions of mitochondria, improved energy metabolism in the cell, and the ability to maintain macroerg levels, including with hypoxia. It activates the intracellular synthesis of protein and nucleic acids, the enzymatic processes of the Krebs cycle, promotes glucose utilization, the synthesis and intracellular accumulation of ATP, restores the structure and functions of membranes, and has a modulating effect on membrane-bound enzymes, ion channels, and receptor complexes. Mexico improves metabolism and blood supply to internal organs, rheological properties of blood and microcirculation, has anti-ischemic properties: improves blood flow, limits the area of ischemic damage and stimulates reparative processes; increases the body's resistance to extreme factors and oxygen-dependent pathological conditions; promotes regression of signs of vasomotor instability. The half-life of the drug ranges from 8-12 hours (Russian Drugs Register of Radar 2008. - M., "Radar - 2008", 2007 - P.538-540).

The invention is illustrated by the figures:

Figure 1. Liver. Infiltration in the area of necrosis. Pressing PDS for 30 minutes. Without the use of drugs. 3 days after ischemia. Coloring: hematoxylin and eosin. Magnification 280 ^x

Figure 2. Liver. Preservation of structure, reduction of glycogen content. Pressing PDS for 30 minutes. The introduction of Mexico and serotonin adipate. 3 days after ischemia. Coloring by McManus. Increasing ^x 500

Figure 3. Liver. Preservation of the structure, the accumulation of glycogen. Pressing PDS for 30 minutes. The introduction of Mexico and serotonin adipate. 14 days after ischemia. Coloring by McManus. Increasing ^x 500.

The method is as follows.

Serotonin adipate diluted in 5 ml of isotonic NaCl solution at a dose of 0.15 mg / kg of body weight is injected intravenously slowly to an experimental animal (dog) 1 hour before the operation; 35 minutes before the operation, Mexico is also slowly injected intravenously in the form of a 5% solution at a dose of 5 mg / kg of body weight. After processing the surgical field with a 1% solution of chlorhexidine bigluconate under intravenous hexenal anesthesia (0.03 g / kg), a laparotomy is performed, the hepatoduodenal ligament is infiltrated with 10 ml of a 0.25% novocaine solution and squeezed by an elastic intestinal clamp for 30 minutes. Then, a resection of the left external lobe of the liver is performed. After the release of the hepatoduodenal ligament, the wound of the anterior abdominal wall is sutured tightly. After 2 hours after the first injection, serotonin adipate, diluted in 5 ml of isotonic NaCl solution at a dose of 0.2 mg / kg body weight, is slowly injected intravenously, and 10 hours after the first administration - at a dose of 0.1 mg / kg body weight . Following this, 11 hours after the first administration of Mexico, Mexicor is injected slowly intravenously in a dose of 7 mg / kg of body weight. In the postoperative period, each of the drugs is administered for 8 days: Mexico - three times a day - at 9 ⁰⁰ at a dose of 2.5 mg / kg of body weight, at 15 ⁰⁰ at a dose of 1.5 mg / kg of body weight, at 21 ⁰⁰ at a dose of 6 mg / kg body weight, serotonin adipate - twice a day - at 8 ³⁰ in a dose of 0.15 mg / kg body weight, at 17 ⁰⁰ in a dose of 0.1 mg / kg body weight.

The study was performed on 18 outbred dogs of both sexes weighing from 9 to 18.5 kilograms at the age of 2 to 5 years. The age was determined by teeth using the method of I.P. Zapadnyuk (Zapadnyuk I.P., Zapadnyuk V.I., Zakharia E.A., Zapadnyuk B.V. Laboratory animals (breeding, keeping, using in the experiment) .- 3 -th ed., revised and additional - Kiev: Vishcha school, 1983.- 384 s). Experimental animals were observed up to and including 30 days; they collected blood from the femoral vein for biochemical studies and a puncture biopsy of the liver to determine the content of lipid peroxidation products (LPO) in the liver tissue homogenate. The determination of the level of transaminase activity of alanine aminotransferase (ALT) and aspartate aminotransferase (ACT) in the blood serum was carried out according to the Reitman-Frenkel method, the concentration of bilirubin - according to the method of Endrashik, cholesterol according to Ilk, β-lipoproteins according to Burstein. In addition, the levels of alkaline phosphatase (ALP), total protein, prothrombin index (IPT), and fibrinogen were determined by standard methods. Indicators of cytolysis syndrome were the activity of ALT and ACT. Indicators of cholestasis syndrome are alkaline phosphatase activity, bilirubin concentration. Indicators of hepatosuppressive syndrome were the concentration of total protein, fibrinogen content, and prothrombin index. The content of lipid peroxidation products - diene conjugates (DCs) and malondialdehyde (MDA) - was determined by the method of I.D. Stalna et al. (1977). Animals were withdrawn from experience only for taking liver tissue samples for morphological studies - pieces of material 1 × 0.5 × 0.5 cm in size were taken. Their processing was carried out according to standard methods with fixation in 10% neutral formalin, dehydration in alcohols of ascending concentration and formation paraffin blocks with the subsequent production of histotopographic sections 5-7 microns thick on a sledge microtome. The medium was stained with hematoxylin with eosin and according to McManus. The drugs were studied at a light-optical level with an increase of 280 ^x -500 ^x . Before the start of the experiments, all animals had blood taken from the femoral vein for biochemical studies (control). Throughout the study, 15 animals were injected with serotonin adipate and Mexicor according to the scheme we proposed. The surgical field was treated with a 1% solution of chlorhexidine bigluconate, under intravenous hexenal anesthesia (0.03 g / kg), a laparotomy was performed, the hepatoduodenal ligament 10 ml of 0.25% novocaine solution was infiltrated, and it was squeezed with an elastic intestinal clamp for 30 minutes. Then, the elements of the glisson and caval systems of the left external lobe of the liver were bandaged, after which the left triangular ligament was crossed. Following this, a resection of the left external lobe of the liver was performed with its blunt and sharp separation along the interlobar sulcus and plastic wound surface with a lock of a large omentum on the leg. After removing the clamp from the hepatoduodenal ligament, the wound of the anterior abdominal wall was sutured tightly. Three dogs underwent clamping of the hepatoduodenal ligament for 30 minutes and resection of the left external lobe of the liver without administration of the studied drugs.

With a 30-minute liver ischemia without pharmacological protection in the postoperative period, animals showed adynamia, lethargy, and lack of appetite, which indicated the severity of changes in the body of dogs. 1 animal died on the 2nd day after surgery, 2 survived to 3 days.

Under the influence of the PDS block lasting 30 minutes against the background of the administration of mexicor and serotonin adipate, the postoperative period was satisfactory, one hundred percent survival of the animals was observed.

In a morphological study of liver tissue after exposure to 30-minute ischemic anoxia without the use of Mexico and serotonin adipate in the first two days of the experiment, plethora of triads and central veins, an increase in the area of hepatic parenchyma with dystrophic and necrobiotic changes, an increase in interstitial edema, and aggravation of hemodynamic were determined . In the central zones of the hepatic lobules and in the zone of triads, common stasis was observed in the portal and central veins, tears of their walls against the background of increasing discomplexation of the lobules themselves. The cytoplasm of hepatocytes was swollen, separate foci of plasmorrhagia were observed. Lipoidosis was combined with granular and hydropic dystrophies with a predominance of the latter. By the 3rd day, discirculatory disorders were growing in the liver tissue - dilatation and plethora of blood vessels, cells with signs of coagulation necrosis were visualized, leukocyte infiltration appeared between the hepatic lobules and beams, there was practically no glycogen in the hepatocytes (Fig. 1).

Although 30-minute occlusion of PDS with the administration of mexicor and serotonin adipate was accompanied by swelling of hepatocytes, and in some areas of the liver parenchyma large-cell infiltrates were found, nevertheless dystrophic and discirculatory disorders were much less pronounced. The picture of further changes and their dynamics confirm this observation. During the first three days of the experiment in hepatocytes, the glycogen content decreased to moderate (figure 2). Subsequently, glycogen reserves were intensively restored. So already during histological examination on the 14th day in the liver cells, a significant amount of glycogen was noted along with individual sections of granular dystrophy (figure 3). Over the next two weeks, the intensive accumulation of glycogen continued and by the 30th day the state of the liver tissue was completely restored.

Without the use of the studied drugs with 30-minute ischemic anoxia of the liver, cytolytic, cholestatic and hepatosuppressive syndromes proceeded simultaneously, as evidenced by the results of biochemical studies. So after 30 minutes of bleeding after half an hour of reperfusion, a significant increase in the digital values of transaminases was noted (table 1; units: ALT and ACT - mmol / (h · l), DC and MDA - mmol / g; p <0.05; n = 18 ) and bilirubin of blood serum (table 2; units: bilirubin - mmol / l, alkaline phosphatase - mmol / (h · l), cholesterol - mmol / l, β-lipoproteins - g / l; p <0.05; n = 18). In the same time interval, the content of alkaline phosphatase also increased (Table 2), simultaneously accompanied by a decrease in the amount of fibrinogen and IPT (table 3; units: fibrinogen - mg / l, IPI -%, total protein - g / l, urea - mmol / l; p <0.05; n = 18). The most pronounced changes in these indicators occurred at the end of the 3rd day from the beginning of the experiment. Compared with the control data, the increase in ALT and ACT values on the third day was 6.98 and 6.93 times, respectively, which indicates the severity of the cytolytic syndrome (table 1).

Table 1 experience conditions time after ischemia (day) ALT ACT DK MDA 1 control - 0.74 ± 0.08 0.46 ± 0.06 0.87 ± 0.081 0.072 ± 0.009 2 liver ischemia 30 minutes 1.95 ± 0.11 1.11 ± 0.04 1.68 ± 0.059 0.142 ± 0.007 3 liver ischemia; mexicor, serotonin adipate 30 minutes 1.16 ± 0.07 0.67 ± 0.09 1.10 ± 0.062 0.096 ± 0.005 4 liver ischemia one 3.44 ± 0.36 1.71 ± 0.17 1.96 ± 0.064 0.191 ± 0.004 5 liver ischemia; mexicor, serotonin adipate one 2.21 ± 0.09 0.96 = 0.08 1.22 ± 0.053 0.125 ± 0.007 6 liver ischemia 3 5.17 ± 0.28 3.19 ± 0.14 2.25 ± 0.069 0.239 ± 0.005 7 liver ischemia; mexicor, serotonin adipate 3 2.32 ± 0.14 1.48 ± 0.11 1.43 ± 0.042 0.130 ± 0.008 8 liver ischemia; mexicor, serotonin adipate 7 1.87 ± 0.15 1.22 ± 0.13 1.12 ± 0.067 0.103 ± 0.006 9 liver ischemia; mexicor, serotonin adipate fourteen 1.12 ± 0.18 0.85 ± 0.07 0.97 ± 0.079 0.097 ± 0.003 10 liver ischemia; mexicor, serotonin adipate thirty 0.73 ± 0.08 0.47 ± 0.11 0.84 ± 0.064 0.074 ± 0.012

table 2 experience conditions time after ischemia (day) bilirubin alkaline phosphatase cholesterol β-lipo
proteids 1 control - 13.46 ± 0.27 2.46 ± 0.29 3.94 ± 0.42 0.32 ± 0.017 2 liver ischemia 30 minutes 22.94 ± 0.83 3.81 ± 0.27 3.87 ± 0.57 0.35 ± 0.026 3 liver ischemia; mexicor, serotonin adipate 30 minutes 16.72 ± 0.34 2.92 ± 0.18 3.93 ± 0.36 0.28 ± 0.019 4 liver ischemia one 35.18 ± 0.47 4.08 ± 0.39 3.64 ± 0.89 0.39 ± 0.031 5 liver ischemia; mexicor, serotonin adipate one 19.94 ± 0.71 3.84 ± 0.19 3.89 ± 0.43 0.34 ± 0.027 6 liver ischemia 3 39.53 ± 0.92 4.96 ± 0.32 2.92 ± 0.13 0.47 ± 0.033 7 liver ischemia; mexicor, serotonin adipate 3 23.96 ± 0.85 2.89 ± 0.27 3.81 ± 0.39 0.33 ± 0.029 8 liver ischemia; mexicor, serotonin adipate 7 18.69 ± 0.69 2.75 ± 0.31 3.72 ± 0.51 0.32 ± 0.019 9 liver ischemia; mexicor, serotonin adipate fourteen 14.87 ± 1.48 2.41 ± 0.21 3.83 ± 0.52 0.36 ± 0.032 10 liver ischemia; mexicor, serotonin adipate thirty 13.51 ± 0.83 2.44 ± 0.17 3.93 ± 0.23 0.31 ± 0.021

Table 3 experience conditions time after ischemia (day) fibrinogen PTI total protein urea 1 control - 3.88 ± 0.18 81.7 ± 2.11 70.1 ± 1.12 3.82 ± 0.29 2 liver ischemia 30 minutes 2.64 ± 0.13 71.9 ± 2.21 68.5 ± 1.92 4.02 ± 0.43 3 liver ischemia; mexicor, serotonin adipate 30 minutes 3.41 ± 0.12 80.8 ± 1.73 69.3 ± 2.03 3.91 ± 0.18 4 liver ischemia one 2.12 ± 0.25 58.7 ± 1.39 61.3 ± 1.29 5.73 ± 0.29 5 liver ischemia; mexicor, serotonin adipate one 3.24 ± 0.17 76.2 ± 1.26 67.4 ± 1.23 4.58 ± 0.33 6 liver ischemia 3 2.03 ± 0.15 44.8 ± 1.28 60.2 ± 1.15 7.51 ± 0.27 7 liver ischemia; mexicor, serotonin adipate 3 2.97 ± 0.19 75.2 ± 1.31 66.2 ± 2.17 4.84 ± 0.32 8 liver ischemia; mexicor, serotonin adipate 7 3.15 ± 0.18 76.9 ± 1.36 68.2 ± 2.34 3.95 ± 0.21 9 liver ischemia; mexicor, serotonin adipate fourteen 3.74 ± 0.19 80.2 ± 1.15 71.3 ± 1.82 3.87 ± 0.29 10 liver ischemia; mexicor, serotonin adipate thirty 3.87 ± 0.21 82.4 ± 1.37 71.7 ± 1.42 3.81 ± 0.33

At the same time, the bilirubin content increased 2.94 times. The post-ischemic 3-day period was also characterized by an increase in alkaline phosphatase values of 2.02 times, β-lipoproteins - by 1.47 times and a decrease in cholesterol concentration by 1.35 times compared with the control. These changes indicate the development of cholestasis syndrome (table 2). Hepatodepressive syndrome at the end of the third day of the postoperative period was manifested by a quantitative decrease in fibrinogen by 1.91 times, IPT by 1.82 and total protein by 1.16 times compared with the initial data. For three days, the urea content remained high with a peak at 3 days after surgery (table 3). The limiting values of lipid peroxidation products were recorded on day 3 with an increase in DC and MDA by 2.59 and 3.32 times, respectively, compared with the control (table 1).

Under the influence of Mexico and serotonin adipate during half-hour occlusion, the PDS of aminotransferases changed less dynamically and with a lower amplitude of digital values, which becomes noticeable after 30 minutes of reperfusion (table 1). Comparing the data obtained on terms similar to the previous series of experiments, but without drug administration, it becomes obvious that the maximum activity of the studied enzymes also falls on the 3rd day of the postoperative period. Although ALT and ACT indices increased 3.14 and 3.22 times, respectively, compared with the control, at the same time they were lower than those obtained with 30-minute liver ischemia without administration of the studied drugs: ALT 2.23 times , ACT - 2.16 times. By the 14th day, the content of ALT, ACT in the blood serum decreased 2.07 and 1.74 times, respectively (on average 1.91 times, that is, almost twice) with full normalization by the 30th day after the operation. In a biochemical study of the bilirubin content (table 2), the highest level in the blood serum was observed on the 3rd day of observation of the operated animals; compared with the control, the increase was 1.78 times. Then, in parallel with transaminases, by 14 days there was a noticeable decrease of 1.61 times relative to the maximum values achieved on the 3rd day. By the end of the experiment, there was no difference with normal values. It should be noted that the level of this pigment was significantly lower than in experiments with 30-minute ischemia without protection with mexicor and serotonin adipate at comparable times. Significant changes in cholesterol, β-lipoproteins, alkaline phosphatase at all periods of observation were not observed (table 2). At the same time, a slight decrease in fibrinogen, PTI, and total protein was observed during the first 7 days after surgery (Table 3). The urea content in the blood serum was characterized by an increase in its level by 1.27 times on 3 days with a gradual decrease to normal by the end of the 14th day (table 3).

The experiments using mexicor and serotonin adipate were characterized by less activation of LPO processes, the level and dynamics of accumulation of products of which were significantly lower compared with data obtained with 30-minute liver ischemia without drug administration (table 1). The increase in the content of malondialdehyde and diene conjugates in the liver tissue 30 minutes after the end of the operation was 1.33 and 1.26 times, respectively. The greatest increase in the content of lipid peroxidation products in the liver tissue homogenate occurred on the 3rd day and amounted to: DC - 1.64 times; MDA - 1.81 times, while at the same time 1.57 and 1.84 times less compared with the group without drug administration. Starting from the 7th day of the postoperative period, there was a downward trend, which became stable over the course of the week, which is confirmed by the following data: compared with 3 days, the decrease in DC and MDA on the 7th day of observation was 1.28 and 1, 26 times, and on day 14 - already 1.47 and 1.34 times, respectively. By the thirtieth day, the studied indicators returned to their original values.

Thus, under the influence of Mexico and serotonin adipate, almost all of the studied parameters, starting from the seventh day, had a pronounced trend of positive dynamics up to the complete normalization of their values by the 30th day of observation.

Examples of specific application.

Example No. 1. Dog No. 3. Operation on October 18, 2010. After treating the surgical field with a 1% solution of chlorhexidine bigluconate under intravenous hexenal anesthesia (0.03 g / kg), an upper middle laparotomy was performed, the hepatoduodenal ligament 10 ml was visualized and infiltrated 0, 25% novocaine solution, squeezed it with an elastic intestinal clamp for 30 minutes. Then, the elements of the glisson and caval systems of the left external lobe of the liver were ligated, and then the left triangular ligament was crossed. Following this, a resection of the left external lobe of the liver was performed with its blunt and sharp separation along the interlobar sulcus, imposition of liver sutures on the organ stump and plastic wound surface with a large omentum on the leg. After removing the clamp from the hepatoduodenal ligament, the wound of the anterior abdominal wall was sutured tightly.

Results of biochemical studies:

10/18/2010 - before the operation: ALT-0.67 mmol / (h · l); AST - 0.41 mmol / (h · l); bilirubin - 13.32 μmol / l; Alkaline phosphatase - 2.19 mmol / (h · l); cholesterol - 4.31 mmol / l; β-lipoproteins - 0.30 g / l; fibrinogen - 3.96 mg / l; IPT - 83.7%; total protein 71.2 g / l; urea - 4.11 mmol / l; DK - 0.74 mmol / g; MDA - 0.067 mmol / g.

October 18, 2010 - after 30 minutes of reperfusion: ALT - 2.19 mmol / (h · l); AST - 1.14 mmol / (h · l); bilirubin - 23.68 μmol / l; Alkaline phosphatase - 4.05 mmol / (h · l); cholesterol - 3.31 mmol / l; β-lipoproteins - 0.36 g / l; fibrinogen - 2.52 mg / l; IPT - 69.7%; total protein - 66.5 g / l; urea - 4.43 mmol / l; DK - 2.08 mmol / g; MDA - 0.146 mmol / g.

10/19/2010: ALT - 3.78 mmol / (h · l); AST - 1.86 mmol / (h · l); bilirubin - 35.51 μmol / l; Alkaline phosphatase - 4.56 mmol / (h · l); cholesterol - 4.28 mmol / l; β-lipoproteins - 0.39 g / l; fibrinogen - 1.98 mg / l; IPT - 57.4%; total protein - 60.1 g / l; urea - 5.98 mmol / l; DK - 1.62 mmol / g; MDA - 0.193 mmol / g.

10.21.2010: ALT - 5.43 mmol / (h · l); AST - 3.31 mmol / (h · l); bilirubin - 40.38 μmol / l; Alkaline phosphatase - 4.23 mmol / (h · l); cholesterol - 2.79 mmol / l; β-lipoproteins - 0.76 g / l; fibrinogen - 1.93 mg / l; IPT - 43.6%; total protein - 59.8 g / l; urea - 7.74 mmol / l; DK - 2.87 mmol / g; MDA - 0.244 mmol / g.

At the end of the third day of the experiment, the animal died.

Example No. 2. Dog No. 5. Operation 26.10.2010, 1 hour before the operation, serotonin adipate, diluted in 5 ml of isotonic NaCl solution at a dose of 0.15 mg / kg; 35 minutes before the operation, Mexicor was slowly injected intravenously in the form of a 5% solution at a dose of 5 mg / kg of body weight. After treating the surgical field with a 1% solution of chlorhexidine bigluconate under intravenous hexenal anesthesia (0.03 g / kg), an upper-mid laparotomy was performed, the hepatoduodenal ligament was visualized and infiltrated with 10 ml of a 0.25% novocaine solution, squeezed with an elastic intestinal clamp of 30 minutes. Then, the elements of the glisson and caval systems of the left external lobe of the liver were ligated, and then the left triangular ligament was crossed. Following this, a resection of the left external lobe of the liver was performed with its blunt and sharp separation along the interlobar sulcus, imposition of liver sutures on the organ stump and plastic wound surface with a large omentum on the leg. After removing the clamp from the hepatoduodenal ligament, the wound of the anterior abdominal wall was sutured tightly. After 2 hours after the first injection, serotonin adipate, diluted in 5 ml of isotonic NaCl solution at a dose of 0.2 mg / kg body weight, was intravenously slowly injected, and 10 hours after the first administration, at a dose of 0.1 mg / kg body weight . Following this, 11 hours after the first administration of Mexico, Mexicor was injected slowly intravenously in a dose of 7 mg / kg of body weight. In the postoperative period, each of the drugs was administered for 8 days: Mexico - three times a day - at 9 ⁰⁰ at a dose of 2.5 mg / kg of body weight, at 15 ⁰⁰ at a dose of 1.5 mg / kg of body weight, at 21 ⁰⁰ at a dose of 6 mg / kg body weight, serotonin adipate - twice a day - at 8 ³⁰ in a dose of 0.15 mg / kg body weight, at 17 ⁰⁰ in a dose of 0.1 mg / kg body weight.

Results of biochemical studies:

10.26.2010 - before the operation: ALT - 0.76 mmol / (h · l); AST - 0.47 mmol / (h · l); bilirubin - 13.51 μmol / l; Alkaline phosphatase - 2.53 mmol / (h · l); cholesterol - 3.93 mmol / l; β-lipoproteins - 0.31 g / l; fibrinogen - 3.76 mg / l; IPT - 79.4%; total protein 71.2 g / l; urea - 3.78 mmol / l; DK - 0.89 mmol / g; MDA - 0.075 mmol / g.

10.26.2010, after 30 minutes of reperfusion: ALT - 1.14 mmol / (h · l); AST - 0.62 mmol / (h · l); bilirubin - 16.44 μmol / l; Alkaline phosphatase - 2.81 mmol / (h · l); cholesterol - 3.92 mmol / l; β-lipoproteins - 0.29 g / l; fibrinogen - 3.56 mg / l; IPT - 78.1%; total protein - 70.5 g / l; urea - mmol / l; DC - 0.97 mmol / g; MDA - 0.094 mmol / g.

10.27.2010: ALT - 2.06 mmol / (h · l); AST - 0.89 mmol / (h · l); bilirubin - 18.15 μmol / l; Alkaline phosphatase - 3.72 mmol / (h · l); hblesterol - 3.81 mmol / l; β-lipoproteins - 0.32 g / l; fibrinogen - 3.19 mg / l; IPT - 77.3%; total protein - 69.1 g / l; urea - 4.27 mmol / l; DK - 1.03 mmol / g; MDA - 0.112 mmol / g.

10/29/2010: ALT - 2.19 mmol / (h · l); AST - 1.27 mmol / (h · l); bilirubin - 23.09 mmol / l; Alkaline phosphatase - 2.76 mmol / (h · l); cholesterol - 3.83 mmol / l; β-lipoproteins - 0.34 g / l; fibrinogen - 2.99 mg / l; IPT - 76.2%; total protein - 67.4 g / l; urea - 4.39 mmol / l; DK - 1.18 mmol / g; MDA - 0.123 mmol / g.

November 2, 2010: ALT-1.63 mmol / (h · l); AST - 1.15 mmol / (h · l); bilirubin - 18.11 μmol / l; Alkaline phosphatase - 2.67 mmol / (h · l); cholesterol - 3.79 mmol / l; β-lipoproteins - 0.33 g / l; fibrinogen - 3.26 mg / l; IPT - 77.4%; total protein - 69.3 g / l; urea - 3.89 mmol / l; DK - 0.92 mmol / g; MDA - 0.098 mmol / g.

November 9, 2010: ALT-0.98 mmol / (h · l); AST - 0.77 mmol / (h · l); bilirubin - 14.12 μmol / l; Alkaline phosphatase - 2.51 mmol / (h · l); cholesterol - 3.86 mmol / l; β-lipoproteins - 0.32 g / l; fibrinogen - 3.77 mg / l; IPT - 80.5%; total protein - 70.9 g / l; urea - 3.64 mmol / l; DK - 0.86 mmol / g; MDA - 0.095 mmol / g.

November 25, 2010: ALT-0.72 mmol / (h · l); AST - 0.46 mmol / (h · l); bilirubin - 13.49 μmol / l; Alkaline phosphatase - 2.45 mmol / (h · l); cholesterol - 3.95 mmol / l; β-lipoproteins - 0.31 g / l; fibrinogen - 3.86 mg / l; IPT - 81.9%; total protein 71.2 g / l; urea - 3.81 mmol / l; DK - 0.85 mmol / g; MDA - 0.073 mmol / g.

The postoperative period was satisfactory.

Example No. 3. Dog No. 12. Operation 09.11.2010, 1 hour before the operation, serotonin adipate diluted in 5 ml of isotonic NaCl solution at a dose of 0.15 mg / kg of weight was injected intravenously slowly. 35 minutes before the operation, Mexicor was slowly injected intravenously in the form of a 5% solution at a dose of 5 mg / kg of body weight. After treating the surgical field with a 1% solution of chlorhexidine bigluconate under intravenous hexenal anesthesia (0.03 g / kg), an upper-mid laparotomy was performed, the hepatoduodenal ligament was visualized and infiltrated with 10 ml of a 0.25% novocaine solution, squeezed with an elastic intestinal clamp of 30 minutes. Then, the elements of the glisson and caval systems of the left external lobe of the liver were ligated, and then the left triangular ligament was crossed. Following this, a resection of the left external lobe of the liver was performed with its blunt and sharp separation along the interlobar sulcus, imposition of liver sutures on the organ stump and plastic wound surface with a large omentum on the leg. After removing the clamp from the hepatoduodenal ligament, the wound of the anterior abdominal wall was sutured tightly. After 2 hours after the first injection, serotonin adipate, diluted in 5 ml of isotonic NaCl solution at a dose of 0.2 mg / kg body weight, was intravenously slowly injected, and 10 hours after the first administration, at a dose of 0.1 mg / kg body weight . Following this, 11 hours after the first administration of Mexico, Mexicor was injected slowly intravenously in a dose of 7 mg / kg of body weight. In the postoperative period, each of the drugs was administered for 8 days: Mexico - three times a day - at 9 ⁰⁰ at a dose of 2.5 mg / kg of body weight, at 15 ⁰⁰ at a dose of 1.5 mg / kg of body weight, at 21 ⁰⁰ at a dose of 6 mg / kg body weight, serotonin adipate - twice a day - at 8 ³⁰ in a dose of 0.15 mg / kg body weight, at 17 ⁰⁰ in a dose of 0.1 mg / kg body weight.

Results of biochemical studies:

November 9, 2010 - before the operation: ALT - 0.75 mmol / (h · l); AST - 0.45 mmol / (h · l); bilirubin - 13.47 μmol / l; Alkaline phosphatase - 2.48 mmol / (h · l); cholesterol - 3.95 mmol / l; β-lipoproteins - 0.33 g / l; fibrinogen - 3.81 mg / l; IPT - 80.4%; total protein - 69.8 g / l; urea - 3.83 mmol / l; DK - 0.86 mmol / g; MDA - 0.074 mmol / g.

November 9, 2010 - after 30 minutes of reperfusion: ALT - 1.15 mmol / (h · l); AST - 0.63 mmol / (h · l); bilirubin - 16.53 μmol / l; Alkaline phosphatase - 2.84 mmol / (h · l); cholesterol - 3.91 mmol / l; β-lipoproteins - 0.30 g / l; fibrinogen - 3.58 mg / l; IPT - 79.6%; total protein - 69.1 g / l; urea - 3.89 mmol / l; DK - 1.06 mmol / g; MDA - 0.095 mmol / g.

November 10, 2010: ALT - 2.14 mmol / (h · l); AST - 0.92 mmol / (h · l); bilirubin - 19.23 μmol / l; Alkaline phosphatase - 3.81 mmol / (h · l); cholesterol - 3.88 mmol / l; β-lipoproteins - 0.37 g / l; fibrinogen - 3.21 mg / l; IPT - 76.9%; total protein - 68.4 g / l; urea - 4.61 mmol / l; DK - 1.13 mmol / g; MDA - 0.121 mmol / g.

12.12.2010: ALT - 2.27 mmol / (h · l); AST - 1.36 mmol / (h · l); bilirubin - 23.84 μmol / l; Alkaline phosphatase - 2.93 mmol / (h · l); cholesterol - 3.85 mmol / l; β-lipoproteins - 0.33 g / l; fibrinogen - 3.02 mg / l; IPT - 76.4%; total protein - 66.9 g / l; urea - 4.92 mmol / l; DK - 1.23 mmol / g; MDA - 0.129 mmol / g.

November 16, 2010: ALT - 1.76 mmol / (h · l); AST - 1.28 mmol / (h · l); bilirubin - 18.83 μmol / l; Alkaline phosphatase - 2.71 mmol / (h · l); cholesterol - 3.68 mmol / l; β-lipoproteins - 0.34 g / l; fibrinogen - 3.19 mg / l; IPT - 79.3%; total protein - 69.6 g / l; urea - 3.92 mmol / l; DK - 1.03 mmol / g; MDA - 0.101 mmol / g.

11/23/2010: ALT - 1.06 mmol / (h · l); AST - 0.82 mmol / (h · l); bilirubin - 14.33 μmol / l; Alkaline phosphatase - 2.43 mmol / (h · l); cholesterol - 3.87 mmol / l; β-lipoproteins - 0.31 g / l; fibrinogen - 3.71 mg / l; IPT - 81.4%; total protein - 71.5 g / l; urea - 3.90 mmol / l; DK - 0.89 mmol / g; MDA - 0.097 mmol / g.

12/9/2010: ALT - 0.71 mmol / (h · l); AST - 0.48 mmol / (h · l); bilirubin - 13.38 μmol / l; Alkaline phosphatase - 2.46 mmol / (h · l); cholesterol - 3.91 mmol / l; β-lipoproteins - 0.33 g / l; fibrinogen - 3.89 mg / l; IPT - 81.8%; total protein - 70.7 g / l; urea - 3.79 mmol / l; DK - 0.86 mmol / g; MDA - 0.072 mmol / g.

The postoperative period was satisfactory.

The results obtained indicate that the use of our proposed method for the prevention and treatment of the effects of ischemic effects on the liver with intravenous administration of mexicor and serotonin adipinate according to our scheme with a 30-minute liver ischemia reduced the intensity and contributed to the normalization of biochemical processes, prevented the violation of the structure of the hepatic parenchyma, development of cytolytic, cholestatic and hepatosuppressive syndromes and associated expressed functional morpho logical changes, thereby preventing the development of ischemic changes in the organ, which significantly reduced the severity of possible liver damage due to oxygen starvation and provided not only a protective, but also a therapeutic effect.

Thus, our proposed method allows us to extend the safe period of liver bleeding to 30 minutes, provides treatment and prevention of ischemic and reperfusion injuries of the liver tissue and its microvasculature, and helps prevent massive bleeding during operations on this organ.

Claims (1)

A method for the prevention and treatment of the effects of ischemic effects on the liver under conditions of temporary shutdown of the liver, including intravenous administration of adipinate serotonin, characterized in that the resection of the liver is performed on the experimental animal, on the day of the operation, Mexicor is injected intravenously slowly in the form of a 5% solution twice - 35 min before surgery at a dose of 5 mg / kg body weight and 11 hours after the first administration at a dose of 7 mg / kg body weight, serotonin adipate diluted in 5 ml of isotonic NaCl solution is administered intravenously comfortably slowly three times - 1 hour before surgery at a dose of 0.15 mg / kg body weight, 2 hours after the first administration at a dose of 0.2 mg / kg body weight, 10 hours after the first administration at a dose of 0.1 mg / kg body weight ; in the postoperative period, mexicor and serotonin adipinate is administered for 8 days: mexicor - three times a day - at 9 ⁰⁰ at a dose of 2.5 mg / kg of body weight, at 15 ⁰⁰ at a dose of 1.5 mg / kg of body weight, at 21 ⁰⁰ at a dose 6 mg / kg body weight, serotonin adipate - twice a day - at 8 ³⁰ in a dose of 0.15 mg / kg body weight, at 17 ⁰⁰ in a dose of 0.1 mg / kg body weight.

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