DE3820840C1 - Aqueous reperfusion solution for reducing the reperfusion damage after acute peripheral vascular occlusion - Google Patents

Abstract

The invention relates to an aqueous perfusion solution in particular for reducing tissue damage after acute peripheral vascular occlusion, containing the following constituents: 50 to 500 mmol/l sodium 50 to 500 mmol/l chloride 1 to 10 mmol/l potassium 0.1 to 5 mmol/l calcium 0.5 to 5 mmol/l magnesium and 0.5 to 5 mmol/l sulphate ions 20 to 150 mmol/l sodium bicarbonate 5 to 100 mmol/l glucose and 0.01 to 10 mmol/l hydroxyethyl starch with an average molecular weight (/Mw) of 20000 to 500000 and a number average molecular weight (/Mn) of 10000 to 100000, characterised in that it additionally has the following active substances: 5 to 50 mmol/l aspartate 5 to 50 mmol/l glutamate 1 to 50 mmol/l trishydroxymethylaminomethane 0.0005 to 5 mmol/l calcium antagonist up to 10 mmol/l radical scavenger where the pH of the solution is 7.8. With the aid of the aqueous reperfusion solution in conjunction with modification of the initial reperfusion it is possible to avoid additional damage on reperfusion after acute peripheral vascular occlusion.

Description

The invention relates to an aqueous reperfusion solution, the after acute peripheral vascular occlusion is used. Under Adherence to controlled reperfusion conditions and under Use of the reperfusion solution according to the invention becomes a Tissue damage due to reperfusion after acute, peripheral Vascular occlusion by modifying the initial reperfusion avoidable.

The acute, peripheral vascular occlusion was first described in 1911 by Labey treated by direct embolectomy; since 1963 therapy of choice indirect balloon embolectomy catheters (Fogarty TJ, Daily PO, Shumway NE, Krippaehne H: Am J Surg 1971: 122, 231-237). Despite successful restoration the current trajectory are morbidity and mortality of this simple, often performed under local anesthesia, relatively high. Depending on the height of the vascular occlusion on the lower one Exremity (femoral or aorto-iliac region) become Mor talent rates of 15-22% are given. The mortality rate after Embolectomy for completely ischemic occlusions significantly larger than that after revascularization due to a chronic arterial occlusive disease (3-4%) or after Amputations due to chronic vascular occlusion (1-2%).  

The main cause of the relatively high morbidity and Mortality after embolectomy is the reperfusion scheme den (post-ischemia syndrome, tourniquet syndrome, Legrain-Cormier-Haimovici syndrome) (Haimovici H .: Surgery 1960: 47, 739-747; Cormier JM, Legrain M: J Chir 1962: 83, 473-488; Larcan A, Rauber G, Mathieu P, Mass P, Calamai M: Presse Med 1965: 73, 1819-1824). The syndrome was first developed by Haimovici in 1960 and later ter also described by Cormier and Legrain. On it a large number of works on the Complications after acute embolectomy in both Humans as well as animals.

The reperfusion caused by the embolectomy of the ischemic tissue causes a massive loss washing lactate, potassium, myoglobin and unspecific toxins that are systemic in patients Cause acidosis, hyperkalemia and myoglobinuria. The reperfused extremity also develops massive edema, which in turn is a hemoconcentration and Caused by hypovolaemia; this results in renal insufficiency zienz and possibly shock. The reperfusion edema caused also gently increases the subfascial pressure (from Normal values around 0.66 kPa up to 5.32 kPa) (Schmidt- Neuerburg KP: Langenbeck Arch Chir 1982: 358, 221-226), d. h, a compartment syn develops drom and finally muscle necrosis.

A similar reperfusion damage is caused by the heart muscle tissue described: the sudden reperfusion with normal blood after acute coronary occlusion also an explosive edema, hemorrhage and Contraction bands that eventually lead to hemodynami deterioration, arrhythmias and death being able to lead. In more recent publications, the  Pathogenesis of reperfusion damage outlined (Buck berg DG: J Thorac Cardiovasc Surg 1986: 92, 483-487; Allen BS, Okamoto F. Buckberg DG: J Thorac Cardiovasc Surg 1986: 92, 621-635, 636-648, 553-563, 564-572, 583-593; Follette DM, Fey K, Buckberg GC, Helly J, Steed DL, Foglia RP, Maloney JV: J Thorac Cardiovasc Surg 1981: 82, 221-238). It turns out that myocardium tissues both structurally even after 6 hr ischemia is also metabolically intact and that the Reperfu damage is not an inevitable consequence of the Revascularization after acute occlusion. He can largely prevent myocardial tissue if the conditions of reperfusion (i.e. especially pressure, temperature and duration) and the zu composition of the reperfusate (i.e. pH, calcium, osmo larity and free radical scavengers) change accordingly be changed.

The present invention was based on the object reperfusion damage in acute peripheral vessels closure by modification of the initial reperfu solution as well as the reperfusion conditions Reduce.

With the help of the present invention it was possible to prove be that with acute peripheral vascular occlusion

• (a) reperfusion damage after acute extremities ischemia occurs,
• (b) checking the conditions the reperfusion and the composition of the reperfusa t causes a reduction of this damage and
• (c) an immediate recovery of the contraction after 4 hr complete ischemia at room temperature (one time span that was previously considered irreversible damage to the Rat leg) is possible if the Re perfusion is carried out in a controlled manner.

The above tasks are solved by an aqueous reperfusion solution, especially for ver reduction of tissue damage after acute peripheral Vascular occlusion, containing the following components:

50 to 500 mmol / l sodium
50 to 500 mmol / l chloride
1 to 10 mmol / l potassium
0.1 to 5 mmol / l calcium
0.5 to 5 mmol / l magnesium and
0.5 to 5 mmol / l sulfate ions
20 to 150 mmol / l sodium hydrogen carbonate
5 to 100 mmol / l glucose as well
0.01 to 10 mmol / l hydroxyethyl starch with an average molecular weight (M _w ) of 20,000 to 500,000 and a number average molecular weight (M _n ) of 10,000 to 100,000,

characterized in that they additionally include the following Active ingredients has:

5 to 50 mmol / l aspartate
5 to 50 mmol / l glutamate
1 to 50 mmol / l trishydroxymethylaminomethane
0.0005 to 5 mmol / l calcium channel blocker
to 10 mmol / l radical scavenger, the pH of the solution being 7.8.

A preferred embodiment of the present Er invention is that the reperfusion solution 0.2 up to 0.3 mmol / l calcium ions, 0.05 to 0.1 mmol / l Hydroxyethyl starch and using 1.5 mmol / l 2-Mercapto-propionylglycine (MPG) as a radical scavenger and 0.0005 mmol / l "Diltiazem" as calcium antagonist contains.  

According to a preferred embodiment of the present the invention is called calcium antagonist diltiazem (cis (+) - 5- (2-dimethylaminomethyl) -2,3,4,5-tetrahydro-- 2- (4-methoxyphenyl) -4-oxo-1,5-benzothiazepin-3-ylace did) used.

While usually the aqueous intravenous solution for reducing reperfusion damage after acute peripheral vascular occlusion contains hydroxyethyl starch with an average molecular weight ( _w ) in the range from 40,000 to 200,000 and an average number ( _s ) in the range of 20,000, a particular embodiment of the present invention Invention characterized in that hydroxyethyl starch has an average molecular weight of about 450,000 and an average molecular number of about 70,000. According to this preferred embodiment of the present invention, the osmolarity of the aqueous reperfusion solution is adjusted such that it is more than 380 mOsm / l and the oncotic pressure is greater than 3.46 kPa.

The present invention shows that even after characterized limb ischemia (4 hr complete ischemia at room temperature) an immediate recovery of the Contraction is possible if the condition the reperfusion and the composition of the reper fusates are carefully controlled. The gift of one 37 ° C warm, modified reperfusates at one Perfusion pressure of 6.65 kPa for 30 min gave itself immediately after 4 hr of complete ischemia win the stimulable contractility, less Tissue edema and less increase in leg volume  than the gift of the standard perfusate on a perfu ion pressure of 13.3 kPa.

The importance of the reperfusion strategy per se the further fate of the ischemically damaged cell len is through the formation of a "Reperfusion Scha dens "in the legs, which after 4 hr com platter ischemia - without checking the composition and condition of initial reperfusion - reperfun be dated.

The reperfusion damage consists of a massive Ge web edema, leg volume, decreased ge stick to high-energy phosphates and inability of the reperfused skeletal muscle, after elec contraction stimulation.

Those obtained with the help of the present invention Data contradict the work done in the Council after 4 hours of complete ischemia an ir Describe and underline reversible damage at the same time the need for a reperfusion strategy to consider, which is another, unnecessary damage damage to the ischemically damaged skeletal muscles cells prevented.

Table 3 shows that the leg after reperfusion 4 hr of complete ischemia is relatively intact. It occurs only a small increase in water content and Volume after ischemia in a humid chamber at 20-22 ° C. This results in skeletal muscle coincide markedly with those of the heart muscle regional ischemia: only slight swelling was in the ultrastructure and the water content of the Tissue described.  

There is no correlation between the tissue Adenosine triphosphate (ATP) levels and the contract ability of the skeletal muscle after controlled Reperfusion. After 4 hours of complete ischemia, the sinks ATP content down to 4.0 ± 3.1 µmol / g protein; Contra ions could not by electrical stimulation are caused. The total content of adenine Na cleotide (TAN) was only moderately reduced. After uncon controlled reperfusion also could not control tractions and the ATP and TAN Salaries remained low. These observations will be through other studies on dormant, ischemic and reperfused skeletal muscles in rats, cats and Dogs confirmed. The results using the available However, the invention differ in that the ATP values after controlled reperfusion to the nor return value. Nevertheless, only Kon tractions up to 24% of the control values (Table 3) to achieve.

The inability to measure the extent of the ATP level Contraction ability after controlled reperfusion Predicting has already been done on the myocardium wrote (Rosary ER, Okamoto F, Buckberg GD: J Thorac Cardiovasc Surg 1986: 92, 488-501), there is contractility even with very low ATP values possible. First the combination of the following prints zipien, d. H. preferred the features of the invention ter embodiments of the present invention suitable for the reperfusion damage after acute peri prevent vascular occlusion.

Composition of the reperfusate

The composition of the reperfusate according to the invention was modified according to the following principles:

• (1) Hyperosmolarity (ie glucose) and normoon conicity to reduce post-ischemic edema. A major aspect of reperfusion damage is massive tissue edema, swelling of the endothelia and thus blockage of the capillaries, which in turn leads to hypoxia. The edema causes a decreased diffusion capacity for permanent substance between the capillaries and the mitochondria. If the edema takes on excessive forms, the flow through the capillaries completely stagnates; this state is referred to as a "no reflow phenomenon". Therefore, reducing tissue edema contributes to cellular survival. However, the exact mechanisms underlying the increase in permeability are largely unknown. The hydroxyl radical appears to be the primary damaging agent and the xanthine oxidase has been regarded as the cause of the free radicals that are formed in the ischemic skeletal muscle.
  Therefore, works have been published which report on the beneficial effects of mannitol and free radical scavengers (Korthuis RJ, Granger DN, Townsley MI, Taylor AE: Circ. Res. 1985: 57, 599-609) with regard to tissue edema. Mannitol not only increases osmolarity but also has radical scavenger properties.
• (2) Add free radical scavengers (e.g. MPG) to counter the cytotoxic effect of free radicals to act. The free radical scavenger properties of MPG was described on the myocardium (Beyersdorf F, room G, Fuchs J, Kraft H, Veit P, Satter P: Drug Res 1987: 37, 142-149; Mitsos SE, Walden KM, Abrams GD, Schork MA, Lucchesi BR: Circ 1984: 70, II-259).
• (3) Reduction of the Ca ⁺⁺ influx by lowering the Ca ⁺⁺ content in the reperfusate (ie 0.3 mmol / l) and the addition of a Ca ⁺⁺ blocker (ie diltiazem) to thereby reduce the Ca ⁺⁺ - Prevent entry into the cell even after recurrence of normocalcaemia in the standard reper fusion. Controlling the calcium in the reperfusate is particularly important since the Ca ⁺⁺ concentration in the cytosol gradually increases during ischemia. This increased Ca ⁺⁺ content in turn increases the cytosolic or membrane-like Ca ⁺⁺ permeability and thus prepares the massive influx of Ca ⁺⁺ into the cell during reperfusion.
  According to the invention, Ca ⁺⁺ was reduced in the reperfusate and diltiazem, a potent Ca ⁺⁺ blocker, was added in order to reduce the part of the reperfusion damage caused by a high calcium content. Part of the muscle contracture that is noticeable after normocalcaemic re-perfusion is due to Ca ⁺⁺ overloading of the ischemic tissue. The Ca ⁺⁺ overload is caused by the insufficient post ischemic energy production, which normal Ca ⁺⁺ homeostasis cannot achieve.
• (4) increase the glucose concentration to the osmo increase larity and anaerobic energy production to initiate at the start of reperfusion. Hypoxia and Stress stimulates glucose uptake on the skeleton muscle. Perfusion solutions with increased glucose con concentrations improve carbohydrate metabolism, which in turn increases the ischemic tolerance of skeletal muscle kels is improved. According to the Reper fusat no insulin added, as this increases the glucose Absorption in skeletal muscle during lack of oxygen not improved.
• (5) addition of amino acid precursors to the cancer cycle Intermediate products (i.e. glutamate and aspartate, or  their alkali metal salts) to make them more effective oxidative metabolism to produce energy range so that the cellular reparation processes and then the mechanical function is guaranteed.
• (6) Counteract acidosis with a buffer to thus an optimal environment for resuming the to create metabolic functions.

Conditions of reperfusion

During controlled reperfusion, the following were: Conditions changed:

• (1) decreased reperfusion pressure (6.65 kPa) to reduce the post-ischemic edema lessen that by reperfusion with systemic Pressure is generated;
• (2) Normothermal (37 ° C) to the metabolic processes during cellular repair optimization;
• (3) Duration of controlled re perfusion (30 min.) to run cellular reparation processes, before the normal flow is restored.

The model of uncontrolled repefusion with the Standard Krebs-Henseleit solution (Krebs H.A., Henseleit K., Hoppe-Seyler's Journal for Physiological Che mie, 1932, 210, pages 33 to 38) after 4 h more complete Ischemia is a simulation of what is in the clinic with thrombolysis or surgical embolecto is carried out with the Fogarty catheter. By The artery can use both methods in the majority of cases are successfully reopened, however the Revascularization Syndrome (i.e. the Reper fusion damage) is not prevented. This re Perfusion damage can be caused by a fasciotomy, possibly even make an amputation necessary.  

Clinical application

The principles of experimentally developed control reperfusion can be performed in the operating room Cannulation of the aorta femoralis communis realized will. The crystalloid solution could be with a constant pressure. The disadvantages technology

• (1) the patient's volume load
• (2) and the fact that the crystalloid solution cannot meet the oxygen needs of the limb.

It is therefore clinically preferred to use a 4: 1 mixture from blood and crystalloid solution, where the patient's already oxygenated arterial blood centrally from the common femoral artery through the additionally given crystalloid solution modified can be. This principle is already clinically in cardiac surgery in the form of blood cardioplegia (Rosary ER, Okamoto F, Buckberg GD: J Thorac Cardiovasc Surg 1986: 91, 428-435).

Controlled reperfusion is in principle in the Ver run of about 10 min. to perform up to 2 hours the reperfusate preferably having a temperature of Should be 35 to 40 ° C. The pressure to be applied the reperfusion solution should be 4 to 16 kPa. When using your own blood, oxygenation is d. H. the enrichment of the blood perfusate mixture in the Usually not required. While a mix of 4: 1 from blood and reperfusate solution preferred is set, there is the possibility of variation according to the invention in blood and reperfusion solution in Ratio of 10: 1 to 1:10 after acute peripheral use the vascular occlusion.

Starting point for the preparation of the aqueous Reperfu solution to avoid reperfusion damage  after acute peripheral vascular occlusion is firm Concentrate, which contains the above mentioned components in the contains respective amounts.

The reperfusion solution according to the invention is preferred wisely made from a concentrate that is a lot times the concentration of the components of the Contains reperfusion solution or parts of the solution.

For the preparation of the aqueous reper according to the invention The above concentrate becomes sterile  dissolved or suspended in demineralized water dated.

Examples of use

61 rats of the Wistar strain (200-400 g body weight important) who follow a standard diet (Hope B.V., Woerden, Holland), were anesthetized with ether and received 500 IU heparin i. v. Both hind legs were Disarticulated and skinned in the hip joint. The Kompart Mentfascien were always spared. Since the artery fe moralis profunda is not designed for rats, exi a large number of collaterals. A femo ral or iliac vascular ligation is therefore not in the Able to produce complete ischemia.

The right hind leg served as a control and the left one was used for the experiment. The artery and femoral vein were dissected and a steel cannula (⌀ 0.5 mm) inserted into the artery. It was paid special attention to larger collaterals not to close with the cannula. The demoralis vein was opened lengthways to ensure adequate outflow to guarantee. In each case 1 ml of the Reperfu solution is injected into the artery around the cannula test.

At the end of each experiment, the contract Ability of the leg tested, the volume measured and biopsies of the soleus muscle for the determination water content and high-energy phos taken from phate. PH measurements were taken hourly Lich carried out. Biopsies have always been carried out by the "ro ten "(slow) soleus muscle because the This muscle type mainly works oxidatively, while the "white" (fastest) muscles more on the anaero ben metabolism are dependent.  

In addition, it was found that the metabolic ver changes during the shortest phases of ischemia in the "red" muscles are more pronounced than in the "white".

Experimental groups

In group I (control), 17 legs were removed immediately Exarticulation examined.

In group II (control perfusion) there were 5 hind legs immediately after cannulation at a pressure of 13.3 kPa perfused. Perfusion was at a 37 degree Celsius warm standard reperfusion solution (Tab. 1) for 60 minutes to check the validity of the Per to examine the merger model.

In group III, 15 legs were diagnosed after 4 hr of ischemia Room temperature (20-22 degrees Celsius) examined, d. H. without reperfusion.

In group IV the embolectomy, i.e. H. the uncon trolled reperfusion simulated. In addition there were 12 legs after 4 hr ischemia with a standard reperfusion solution (37 degrees Celsius) at a perfusion pressure of 13.3 kPa perfused for 60 minutes.

Group V was used to simulate the controlled reper fusion. After 4 hr ischemia, 12 hind legs were seen with a reperfusate according to the invention (Tab. 1) in a Perfusion pressure of 6.65 kPa for 30 min. at 37 degrees Celsius perfused. After that, these were Legs another 30 min. with the Krebs-Henseleit solution perfused at 13.3 kPa before the above mes solutions were carried out.  

Perfusion device

After attaching the disarticulated hind leg, the arterial cannula was connected to the tube system of the perfusion device. The hoses were impermeable to gas. The perfusion solutions were filled in 2 double-walled glass cylinders. These cylinders were located 68 cm and 136 cm above the arterial cannula in order to achieve a flow-independent, constant perfusion pressure of 6.65 kPa and 13.3 kPa, respectively. The perfusates were constantly gassed with a mixture of 95% O₂ and 5% CO₂ (Carbogen®), (Messer-Griesheim, Frankfurt / M., FRG); a pO₂ of 78.5 ± 3 kPa (n = 10) was achieved. The temperature of the solutions was thermostatted at 37 ° C. and the tubes were insulated with aluminum paper. The perfusates were passed through blood transfusion filters before they reached the femoral artery. The venous effluate was collected in a separate container and the amount every 2 min. read. The solution was not recirculated.

Perfusion solutions

A standard reperfusate, the so-called Krebs-Henseleit solution (Table 1), the additionally 6 g / 100 ml of hydroxyethyl starch (HES) ent held. If the perfusion of control legs (without Ischemia) with the standard solution without an oncotic effective substance was carried out, a Increase in water content. The invention before HES had a molecular weight average _w from 450,000 with a number average molecular weight _n (M_n) from 70,000 (Fresenius, Oberursel, FRG) and that is why albumin(Mn 69,000) comparable. The osmotic pressure  was carried out with a "semi-micro-osmometer" (Knauer, Bad Homburg, FRG) measured.

In the reperfusate according to the invention, the glucose concentration, the calcium content and the pH value vari iert; aspartate, glutamate, diltiazem and 2-mercapto-propionylglycine (MPG) added.

The exact composition of both solutions is in Table 1 shown.

MPG was added in a concentration of 1.5 mmol / l because previous dose-response studies (with conc ratios between 0.2 and 5 mmol / l) best functionality le results (in cardiac muscle tissue) at a conc achieved from 1-1.5 mmol / l.

Hind leg stimulation

A weight of 1.5 g was attached to the pastern ankle always attached. Pilot studies showed that none Difference in the amplitude of the resulting Contraction revealed whether the electrical stimulation was on thigh muscles or on the sciatic nerve was led. Therefore, in all experiments the Muscles stimulated directly. Rectangular pulses from one electrical stimulator (Stimi, Hugo Sachs Co., book home, FRG) with a stimulation frequency of 1 Hz, stimulus duration of 1 ms and a voltage of 4-8 V were pro Measured 4 times. The Darge in the results set value is the mean of these 4 measurements. The amplitude of the isotonic contraction (in mm) against the weight of 1.5 g was measured. Every leg served as his own control value. The values after the reperfusion was as a percentage of the control con tractions (% KK) expressed. To the optimal chip  Finding voltage and duration of stimulus were made during pilot study the stimulus duration from 0.1 msec to 5 msec, and the Voltage changed from 1 V to 20 V. It was found, that the best results after electrical stimula tion of 8 V and 1 msec. The stream strength could not change with the device used be changed.

Flow

The volume of the effluate was increased every 2 min. measured.

Volume of the hind leg

The leg was placed in a Krebs-Henseleit solution filled vessel and the displaced volume measured. The data is as a percent of the control values of each leg after disarticulation (% KV) specified.

Water content

Biopsies (2-3 g) were taken with a scalpel The soleus muscle obtained, put on wax paper and in a drying cabinet (100 ° C) for 24 h constant weight dried. The water content of the Tissue was calculated using the following equation:

High energy phosphates

Adenosine triphosphate (ATP) was obtained from biopsies of the Soleus muscle with the luciferin / luciferase reaction  (Lundin A, Rickardsson A, Thore, A: Anal Biochem 1976: 75, 611-620).

This very sensitive luminometric method is able to determine ATP values down to 10 ^-15 mol. The luciferin / luciferase reagent (ATP-Monitoring Reagent, LKB Wallac) results in the emission of light of almost constant intensity, which is proportional to the ATP content and is recorded by a luminometer (LKB Wallac 1250). The total nucleotide content (TAN) was calculated by adding ATP, adenosine diphosphate (ADP) and adenosine monophosphate (AMP).

The Energy Charge (EC) is a parameter and indicates to what extent the ATP-ADP-AMP system with high energy Phosphate groups is filled. EC is through the following Equation calculated:

Control values have been given as 0.85 (Atkinson DE: Biochemistry 1968: 7, 4030-4034).

pH measurements

The intramuscular pH was determined by introducing a precalibrated, rustproof beam electrode (21 G, In gold, FRG) in the soleus flexor digitorum muscle superficialis measured (O'Donnell TF: Lancet 1975: 20,533). The pre-calibrations were carried out with standard pH Solutions (pH 4 and pH 7) carried out. Measurements were shortly after the amputation and every hour thereafter with the help of a pH meter (digital pH meter, kink, FRG).  

statistical evaluation

The data are expressed as the mean ± standard deviation (SD) specified. The comparison between the groups was carried out with the Wilcoxon test. differences were indicated as statistically significant from p <0.05 ben.

Comparative Example 1 (Group I)

17 legs were examined immediately after disarticulation. The water content was 75.0% ± 0.9% of the intramuscular The pH reached 7.1 ± 0.1.

Isotonic contractions of 10.5 ± 2.3 mm could from the hind legs.

The high-energy phosphates were in the normal range (Ta belle 2).

Comparative Example 2 (Group II)

5 legs were oxygenated with standard cancer hense conductive solution (Table 1) 60 min. at a Perfusion pressure of 13.3 kPa perfused (without Ischemia). Compared to the values immediately after Disarticulations were not Differences in volume (97.6 ± 2.5% vs 100% KV), Water content (76.7 ± 0.5 vs. 75.0 ± 0.9%) or pH value (7.2 ± 0.2 vs 7.1 ± 0.1). The data for ATP (27.1 ± 13.4 vs 26.2 ± 15.4 µmol / g protein), TAN (33.3 ± 1.4 vs 37.7 ± 17.0 µmol / g protein) and EC (0.84 ± 0.09 vs. 0.73 ± 0.10) were also in the Normal range. The flow was 4.3 ± 0.8 ml / 100 g / min.  

Comparative Example 3 (Group III)

After 4 h of ischemia without reperfusion, no con traction triggered by electrical stimulation and the intramuscular pH dropped from 7.1 ± to 6.3 ± 0.1 (p <0.001). Furthermore, an si Significant decrease in ATP (4.0 ± 3.1 vs. 26.2 ± 15.4 µmol / g protein, p <0.001), of the TAN (15.1 ± 7.7 vs. 37.7 ± 17.0 µmol / g protein, p <0.05) and des EC (0.39 ± 0.08 vs. 0.73 ± 0.10, p <0.01).

In volume (103.6 ± 2.4% KV) and water content (78.7 ± 0.9 vs. 75.0 ± 0.9%, p <0.01) occurred only slightly Increase on (Table 3).

Comparative Example 4 (Group IV)

After 4 h of ischemia and 60 min of uncontrolled reper fusion, only 2 of 12 legs showed minimal contractions after stimulation (2.3 ± 5.5% KK). The volume increased to 121.5 ± 10.2% KV (p <0.001), the water content increased significantly (83.0 ± 2.0 vs. 75.0 ± 0.9%, p <0.001) ( Fig. 1). The flow, which was constant during the 60 min perfusion, decreased to 3.9 ± 0.5 ml / 100 g / min. The high-energy phosphates were still significantly reduced and did not differ from the values after ischemia (Table 2).

Example (group V)

By checking the initial reperfusion as above described, could in connection with the fiction immediate reperfusion solution according to Table 1 Contraction can be achieved in all legs (24.3 ± 10.3% KK). In addition, the water content (76.9 ± 2.4 vs. 75.0 ± 0.9%) and the high energy Phosphates as well as TAN and EC in the normal range (Table 3).  

In contrast to uncontrolled reperfusion (Ver same example 4) continued to occur a significant ge less volume increase (105.0 ± 5.3 vs. 121.5 ± 10.2% KV, p <0.05), d. that is, the leg volume was in Range of the norm remained.

The flow was during the 30 min reperfusion 6.65 kPa 2.1 ± 0.4 ml / 100 g / min and increased to 3.6 ± 0.3 ml / 100 g / min during the second 30 min at one Reperfusion pressure of 13.3 kPa too. The intramuscular pH remained significantly lower (6.3 ± 0.1 vs. 7.1 ± 0.1, p <0.01).

Table 1

Composition of the standard reperfusion solution and the modified reperfusate according to the invention (in mmol / l)

Table 2

Volume, water content, intramuscular pH and high-energy phosphates in controls and control perfusion

Table 3

Volume, water content, intramuscular pH, function and high-energy phosphates of the control group, after 4 h of complete ischemia, after uncontrolled and controlled reperfusion in the rear of the rat

Claims (3)

1. Aqueous reperfusion solution, in particular for reducing tissue damage after acute peripheral vascular occlusion, containing the following components: 50 to 500 mmol / l sodium
50 to 500 mmol / l chloride
1 to 10 mmol / l potassium
0.1 to 5 mmol / l calcium
0.5 to 5 mmol / l magnesium and
0.5 to 5 mmol / l sulfate ions
20 to 150 mmol / l sodium hydrogen carbonate
5 to 100 mmol / l glucose as well
0.01 to 10 mmol / l hydroxyethyl starch with an average molecular weight ( _w ) of 20,000 to 500,000 and a number average molecular weight ( _s ) of 10,000 to 100,000, characterized in that it additionally has the following active ingredients: 5 to 50 mmol / l aspartate
5 to 50 mmol / l glutamate
1 to 50 mmol / l trishydroxymethylaminomethane
0.0005 to 5 mmol / l calcium channel blocker
to 10 mmol / l radical scavenger, the pH of the solution being 7.8. 2. reperfusion solution according to claim 1 with a Content of 30 mmol / l glucose, 0.2 to 0.3 mmol / l Calcium ions and 0.05 to 0.1 mmol / l hydroxy ethyl starch and using 1.5 mmol / l 2-Mercaptopropionylglycine as a radical scavenger and from 0.0005 mmol / l "Diltiazem" as calcium antagonist.   3. reperfusion solution according to claim 1 or 2, characterized characterized in that the hydroxyethyl starch average molecular weight _w) of about 450,000 and a number average molecular weight( _n) from about 70,000 and the osmolarity of the solution above 380 mosm / l preferably 480 mOsm / l and the oncotic pressure larger than 3.46 KPa.