Method for renal transplantation

FIELD: medicine, renal transplantation.

SUBSTANCE: one should wash donor's kidney with cooled solution up to +10 - +8° C. Then one should apply electrodes onto the upper and lower renal poles to perform its single electrostimulation simultaneously with including renal transplant into recipient's general circulation with impulses of 5-=10 msec and sequence frequency being 1 Hz and current power of about 40-50 mcA at seance duration being about 12-15 min. The innovation enables to prevent ischemia occurred in the course of transplantation.

EFFECT: higher efficiency.

1 ex, 2 tbl

 

The present invention relates to medicine, in particular to transplantation and may be used to generate experimental method of preparation of donor kidneys for transplantation with subsequent clinical application.

The known method of transplantation of kidneys, which consists in the fact that the donor kidney was washed with special chilled solution, remove a kidney to the recipient, where it is stitched to the renal vessels of the donor kidney with subsequent stimulation of the renal nerves (1).

However, in this method there are complications in the early postoperative period, and not fully recovers the function of the kidney transplant.

The aim of the invention is to prevent complications and restore full function of the donor kidney.

This objective is achieved in that the electrodes are located on the upper and lower poles of the kidneys, and the electrical stimulation is carried out simultaneously with the inclusion of kidney transplant in the General blood recipient pulses of 5-10 msec and a repetition rate of 1 Hz at an amperage of 40-50 MCA duration of the session 12-15 minutes

A distinctive feature is that a kidney donor washed with a solution "Eurocollins" (USA), cooled to +10-+8°C for 30-40 min, electrodes placed on the upper and lower poles and electrostim is aciu carry disposable with certain parameters on renal graft into the General bloodstream of the recipient.

The proposal is as follows.

Under conditions of General anesthesia in purebred dogs make the fence both donor kidneys, put them on the tray, kanyoro vessels and through the renal artery wash solution "Eurocollins (USA) for 30-40 min at a temperature of +10-+8°C. After washing and cold preservation on the upper and lower poles of one of his kidneys to impose the electrodes.

Then the recipient - mongrel dogs after removal of the native kidneys in their place are donor transplantation. One of them is the control and without electrical stimulation.

Simultaneously with the insertion of the right and left grafts into the General bloodstream provide disposable electrical stimulation pulses of 5-10 MS with a repetition rate of 1 Hz, amperage 40-50 μa at session length 12-15 minutes

The proposal is confirmed by the following example of implementation of the method.

Example 1. Mongrel dog weighing 16 kg - donor and mongrel dog weighing 21 kg of the recipient.

Under conditions of General anesthesia in experimental animal was carried out by the fence of both the donor kidney was placed on the tray, was Coulibaly vessels and through the renal artery was washed with a solution "Eurocollins" for 30 min at a temperature of +10°C. After washing and cold preservation on the upper and lower poles of one of the kidneys of ele is ivali electrodes.

Then the recipient simultaneously transplanted two kidneys: right place remotely experienced with electrical stimulation and to place the left - control. Simultaneously with the inclusion of the right renal transplant into the General bloodstream of the recipient using the "Reflex-3-01" was carried out by electrical stimulation single pulse 10 MS with a frequency of 1 Hz at an amperage of 50 µa duration of the session 15 minutes

Monitoring of transplanted grafts was carried out visually by the color and appearance of the kidneys, histological studies, the dynamics of changes in the activity of redox enzyme (NADH-DG), the histogram of the activity of NADH-DG in the epithelial cells, proximal tubules cells and the results diuretic functions.

The duration of ischemia, seizures kidney was 50 minutes

Urinary function pre electrostimulating kidney was observed after 14 min after implantation, whereas the intact kidney (control) start to operate only after 32 minutes after the operation.

After 15 min and 1 hour after stimulation of stimulated and estimulando kidney was carried out fence biopsy.

It is established that in stimulated and estimulando the grafts, as after 15 min and 1 hour after start-up to blood, watched the same type of p is sticenicima changes in the form of a moderately pronounced swelling interstitielle fabric, spastic contraction of small arteries, spadine glomerular capillary and a lack of red blood cells, moderate necrobiotic changes in tubular epithelium, predominantly the proximal convoluted tubule. However stimulated in the kidney these changes are less pronounced. In addition, there are signs of recovery of microcirculation in the presence of blood in the Lumina of small blood vessels and capillaries. Using enzymatic and chemical reactions it was found that after stimulation of the activity of LDH and NADH-DG in all parts of the nephron is increased in 2 times in comparison with the level of activity of the same enzymes in estimulando the transplant.

The same pattern of increasing activity of redox enzymes can also be seen in a more prolonged ischemia withdrawal of donor kidneys.

In stimulated transplant later, 1 hour after turning it into the bloodstream of the recipient was found granular and loose types of loss of the products of enzymatic and chemical reactions in the cytoplasm of tubular cells of the epithelium, which is typical for deep necrobiotic and dystrophic changes.

It is also shown that the intensity of the luminescence subcapsular cortical layer of the substance restimulating transplant less than stimulated the CSO (310,7 and USD 329.8 used respectively), which is obviously related to the shunting of medullary blood flow and ischemia of the cortical substance in the first case.

It is noted that the intensity of the brain and the cortical substance in estimulando kidney differs significantly less (234,95 and 269,58 used)than in stimulated body (194,45 and 235,85 used).

The intensity of the histological sections of the tissue restimulating transplant was 129,67 used, and stimulated - 58,49 used

In the blood, flowing from restimulating transplant, the concentration of the fluorochrome ranged from 35 used, and flowing from the stimulated kidney - 54 usled

Thus, this example shows the positive impact impact of electrical stimulation on microcirculation and trophism of the parenchyma of the kidney transplant.

The importance of ischemia seizures in the Genesis of the early post-transplantation anuria allogeneic kidney transplant convincingly show the results of a comprehensive morphological studies in experiments with electrical stimulation of the transplanted kidney. The dog - recipient simultaneously transplanted two allogeneic kidney at a place remote from her both kidneys. The donor kidney was prepared for transplantation as well as in renal transplantation person. The duration of ischemia, seizures amounted to 40-50 minutes normally,the urinary function of the grafts did not start immediately after the start of the flow, after 15 minutes But the stimulation of one of the grafts was eliminated rapid acceleration of its excretory function. After 15 min and 1 h after the stimulation of stimulated and estimulando kidney took biopsies for conventional histological examination, histoenzymatic study with subsequent flow cytometric quantification of the activity of redox enzymes: LDH, NADH-DG. In addition, after removing the second biopsy (later, 1 hour after stimulation) in each kidney was introduced through the arterial anastomosis in 1 ml fluorochrome (eurotopia) in phosphate buffer at pH of 7.6 (breeding 25:100000). After 20 min both graft was removed for determination of the concentration and distribution of fluorochrome using the contact of the fluorescent microscope interlocked with the computer. At the same time from venous anastomoses took portions of blood for determination of the concentration of the fluorochrome on spectrofluorimeter No. 224 (England).

Microscopic examination using a standard light microscope in stimulated and estimulando the grafts as after 15 min and 1 hour after start-up to blood, watched the same type of postischemic changes in the form of a moderately pronounced edema of the interstitial tissue, spastic reduce the value of small arteries, spadine glomerular capillary and a lack of red blood cells, moderate necrobiotic changes in tubular epithelium, predominantly the proximal convoluted tubule. In addition, there are signs of recovery of microcirculation in the presence of blood in the Lumina of small blood vessels and capillaries. Using enzymatic and chemical reactions it was found that after stimulation of the activity of LDH and NADH-DG in all parts of the nephron is increased in 1,5-2 times compared with the level of activity of the same enzymes in estimulando the graft (table 1). However, 1 h after stimulation, the levels of enzyme activity in stimulated and estimulando kidneys can converge mainly due to increased activity in restimulation the transplant.

Table No. 1
KidneyTime of withdrawal of the biopsy after stimulationThe activity of the enzyme
the glomeruliproximal tubulesdistally tubules
Stimulated15 minutes0,15±0,020,76±0,090,8±0,12
Nstimerevent0,08±0,01 0,30±0,050,25±0,08
Stimulated1 hour0,78±0,010,78±0,10,46±0,09
Nstimerevent0,11±0,030,59±0,070,4±0,09

The same pattern of increasing activity of redox enzymes can also be seen in a more prolonged ischemia withdrawal of donor kidneys, although less pronounced than in the normal conditions of the experiment (table 2).

In stimulated transplant later, 1 hour after turning it into the bloodstream of the recipient was found granular and loose types of loss of the products of enzymatic and chemical reactions in the cytoplasm of tubular cells of the epithelium, which is typical for deep necrobiotic and dystrophic changes.

Table No. 2

Changes in the activity of NADH-DG failure of conservation of dog kidney
KidneyTime of withdrawal of the biopsy after stimulationThe activity of the enzyme
the glomeruliproximal tubulesthe distal tubules
Stimulated15 minutes 0,11±0,020,72±0,050,58±0,05
Nstimerevent0,09±0,010,61±0,020,29±0,02
Stimulated1 hour0,09±0,020,22±0,030,47±0,03
Nstimerevent0,09±0,010,34±0,010,22±0,02

Revealing the results of contact-fluorescent microscopy. The average of the respective scanogram outer surfaces of the kidneys show that the intensity of the luminescence subkapsuliarnae layer of the cortical substance restimulating transplant less than stimulated (310,7 and USD 329.8 used respectively), which is obviously related to the shunting of medullary blood flow and ischemia of the cortical substance in the first case.

This is confirmed by the results of separate measurements in the area of the brain and the cortical substance on the surface of the sagittal section of the same kidney. It is noted that the intensity of the brain and the cortical substance in estimulando kidney differs significantly less (234,95 and 269,58 used)than in stimulated body (194,45 and 235,85 used). It is quite natural that with the active blood flow in microcirculatory vessels and glomerular capillaries, sledovatel is but and with the active reabsorption of primary urine fluorochrome, the last in a small amount should remain in the cortical substance of the kidney. Lower quantitative indicators of the intensity of luminescence, apparently, also explains the active blood flow and excretion of urine fluorochrome from flowing but the body of blood. Most clearly seen in the measurement of the secondary luminescence of histological sections made from biopsies taken 1 hour after injection of fluorochrome in streams to the transplant blood. The intensity of the histological sections of the tissue restimulating transplant was 129,67 used, and stimulated - 58,49 used

Spectrophotometric study of blood samples from the jugular vein grafts from venous blood, also showed higher content of fluorochrome in venous blood stimulated kidneys.

It is especially clear that the difference was expressed in blood samples taken after 15-20 min after stimulation: in the blood, flowing from restimulating transplant, the concentration of the fluorochrome ranged from 35 used, and flowing from the stimulated kidney - 54 usled

Thus, these data indicate significant effects of heat and cold primary ischemia withdrawal on trophic and moscavide is sustained fashion function allogeneic graft in the early postoperative period. Just conducted 12 experiments in mongrel dogs weighing 14-23 kg Shown a positive effect of the impact of electrical stimulation on microcirculation and trophism of the parenchyma of the kidney transplant. This accordingly allows to reduce the possibility of a primary non-functioning kidney transplants, which has a huge economic effect.

LITERATURE

1. Anti-ischemic protection of the kidneys in conservation and in the post-transplantation period. 1987, abstract. MD, p.88-91. Yudin GV

2. The functional state of a reflex apparatus for deep cooling. Fiziol. log them. Sechenov, 1961, No. 3, s-366. Alishev NV

How kidney transplant by laundering donor kidney cooled solution, a kidney transplant recipient and its stimulation, characterized in that the temperature of the donor kidney is reduced to +10 - +8°s, then the electrodes are placed on the upper and lower poles of the kidneys and electrical stimulation by a single and simultaneous with the start of kidney transplant in the General blood recipient pulses of 5-10 MS with a repetition rate of 1 Hz, amperage 40-50 μa at session length 12-15 minutes



 

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