Method of pyelonephritis prevention in experiment

FIELD: medicine.

SUBSTANCE: invention relates to experimental medicine and concerns pyelonephritis prevention in experiment. During 10 days preparation "Ultrasorb" is introduced to experimental animals with food in terms of 3 g/day. Then pyelonephritis is modelled by rectal infection with uropathogenic strain of lactose-negative E.Coli at the background of cold stress.

EFFECT: absence or reliable reduction of kidney tissue inflammation in animals which obtained "Ultrasorb".

4 ex

 

The invention relates to medicine, namely to Nephrology and experimental medicine, and can be used for the prevention of pyelonephritis in experimental animals (white rats) and develop a way to prevent exacerbation of pyelonephritis in humans.

In the available scientific and patent literature methods of prevention of pyelonephritis in vivo was not found.

The invention is directed to solution of the problem: create a method of prevention of pyelonephritis in experimental animals, resulting from migration of intestinal autoflora into the systemic circulation as a result of acute stress on the background of violations of microbiocenosis of intestines due to improper feeding animals (violation-mode power supply, replacement in the diet of some other products, lack of vitamins in the diet) or artificially created intestinal dysbiosis (Berdichevsky, B.A. et al., 1993, Wmmintrin.h, 2007). You can use a method to prevent acute pyelonephritis patients.

Effect: prevention of pyelonephritis in experimental animals to develop a way to prevent exacerbation of pyelonephritis in humans.

These objectives are achieved by prophylactic use in experimental animals (white rats) t is the within 10 days of preparation "Ultrasorb" at the rate of 3 g/day, mixing it with food. Biologically active additive "Ultrasorb" issued by CJSC "Berry", Kirov, Dugino. The drug is used as a preventive measure practically healthy people when exposed to adverse environmental and production factors, with the increased risk of the formation of goiter, cancer, diabetes, immune deficiency. Also the drug is used alone or as a part of complex therapy in diseases of the gastrointestinal tract (stomatitis, dysbiosis, acute intestinal infections, ulcer of stomach and duodenum, gastritis, cholecystitis, hepatitis, enteritis, colitis), metabolic disorders (atherosclerosis, obesity, diabetes, hyperglycemia), alcohol intoxication and alcoholic pathology in inflammatory pathologies of the skin from bacterial and allergic origin in the complex therapy of secondary immune deficiency.

For the prevention of pyelonephritis this drug is not used.

The main component of "Ultrasorb are fermented insoluble and soluble dietary fiber, which helps to normalize the intestinal microflora, which in turn leads to the strengthening hepatointestinal barrier and prevents tra is socaciu conditionally pathogenic flora of the intestine into the bloodstream.

The method is as follows. The drug "Ultrasorb" after mechanical grinding in a mortar and added to feed for experimental animals (white rats) in an amount of 0.1 (10%) of the daily volume of food (30 g/day per animal, according to the existing norms) 3 g/day. Prophylactic use of the drug "Ultrasorb" is 10 days.

The experiment was performed at 25 animals male and female outbred white rats four months of age contained in standard vivarium conditions. All experiments were conducted in accordance with "the Rules of work with the use of experimental animals" (Annex to order of the Ministry of health of the USSR from 12.08.1977, N755) and the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes" dated March 18, 1986

Animals were divided into 5 experimental groups: control group 2 experimental group not receiving treatment, group 2, treated as an experimental drug therapy "Ultrasorb". Pyelonephritis in animals of the experimental groups was modeled by rectal infection uropathogens lactose-negative strain of Escherichia coli with subsequent single stress exposure in the form of acute cold stress. To infect animals were used in the days the s broth culture uropathogenic strain of Escherichia coli at concentrations of 10 9and 106CFU/ml determined values method.

Group I - control (intact animals), n=5;

Group II - infected Escherichia coli at a concentration of 109CFU/ml with subsequent stress exposure in the form of acute cold stress, n=5;

Group III - infected Escherichia coli at a concentration of 106CFU/ml, also followed by cold stress, n=5;

Group IV - infected Escherichia coli at a concentration of 109CFU/ml, cold stress, giving the drug "Ultrasorb" within 10 days prior to infection and during the subsequent experiment, n=5;

Group V - infected Escherichia coli at a concentration of 106CFU/ml, cold stress, giving the drug "Ultrasorb" within 10 days prior to infection and during the subsequent experiment, n=5.

All experimental animals treated with prophylactic drug "Ultrasorb" and subjected to rectal infection uropathogenic Escherichia coli, and then acute cold stress (groups IV and V)were not significant changes in the peripheral blood during the experiment. When conducting bacteriological studies of autopsy material culture of Escherichia coli, identical in properties taken for rectal infection with the strain was isolated in a significantly smaller percentage of cases compared to animals infected with the rectal same strain, but not receiving "Ultrasorb" (P<0.001). When carrying out histological studies of autopsy material in animals treated with prophylactic treatment "Ultrasorb", inflammatory changes in the kidneys were expressed minimal or absent, whereas in kidneys of animals not receiving "Ultrasorb", attended expressed signs of pyelonephritis: limfomonotsitoza infiltration of the renal pelvis and interstitial kidney, degenerative changes in the tubules.

Examples of specific performance.

Example 1. The female white rats weighing 130 g for 10 days in a mixture with feed gave the drug "Ultrasorb" in the amount of 3 g/day. On the 11th day of the experiment under ether anesthesia the animal rectal introduced 500 l of bacterial suspension daily broth cultures of clinical isolates uropathogenic lactose-negative Escherichia coli at a concentration of 106CFU/ ml On the next day after infection, the animal was subjected to a single stress exposure - cooling to t=+4C for two hours.

Subsequent studies of peripheral blood of the animal, held on the 3rd and 7th days after rectal administration culture, revealed no significant deviations of parameters from baseline. When conducting bacteriological studies of autopsy material of the seed of the studied organs not received the. When conducting histological examination revealed no inflammatory infiltrates in the interstitium and renal pelvis.

Example 2. The female white rats weighing 146 grams for 10 days in a mixture with feed gave the drug "Ultrasorb" in the amount of 3 g/day. On the 11th day of the experiment under ether anesthesia the animal rectal introduced 500 l of bacterial suspension daily broth cultures of clinical isolates uropathogenic lactose-negative Escherichia coli at a concentration of 109CFU/ ml On the next day after infection, the animal was subjected to a single stress exposure - cooling to t=+4C for two hours.

Subsequent studies of peripheral blood of the animal, held on the 3rd and 7th days after rectal administration culture, revealed no significant deviations of parameters from baseline. When conducting bacteriological studies of autopsy material culture of Escherichia coli, the properties are identical to those used to infect strain, was isolated from the kidneys, spleen and liver of rats at a concentration of 102CFU/g When carrying out histological studies revealed scant inflammatory infiltration in the mucosa and stroma renal pelvis.

Example 3. The female white rats weighing 150 g for 10 days in a mixture with feed gave the drug "Ultrasorb" in the amount of 3 g/day. the as of the 11th day of the experiment under ether anesthesia the animal rectal introduced 500 l of bacterial suspension daily broth cultures of clinical isolates uropathogenic lactose-negative Escherichia coli at a concentration of 10 9CFU/ ml On the next day after infection, the animal was subjected to a single stress exposure - cooling to t=+4C for two hours.

Subsequent studies of peripheral blood of the animal, held on the 3rd and 7th days after the rectal Vvedenie culture, revealed no significant deviations of parameters from baseline. When conducting bacteriological studies of autopsy material of the seed of the studied organs not retrieved. When conducting histological examination revealed no inflammatory infiltrates in the interstitium and renal pelvis.

Example 4. Anesthetized with ether female white rats weighing 140 g introduced rectally once with 500 l bacterial suspension daily broth cultures of clinical isolates uropathogenic lactose-negative Escherichia coli at a concentration of 10 CFU/ ml On the next day after infection, the animal was subjected to a single stress exposure - cooling to t=+4C for two hours.

3 days after exposure to stress in the peripheral blood of the experimental animal found a significant increase of leukocytes, continuing until the conclusion of animal experiment. On the 10th day the animal was killed under ether anesthesia by transection of the spinal cord, conducted bacteriological and histolo the systematic examination of the internal organs.

When conducting bacteriological studies of autopsy material culture of Escherichia coli, the properties are identical to those used to infect strain, was isolated from the kidneys, spleen and liver of rats at a concentration of 2*106CFU/g When conducting pathological studies of the colon and small intestine in the stroma of the villi revealed infiltration by lymphocytes, neutrophils and macrophages, with a predominance of lymphocytes. In the kidneys of experimental animal found infiltration interstice by leukocytes, lymphocytes and macrophages, the plethora of vessels, granular degeneration of the epithelium of the tubules and protein cylinders in their gaps, inflammatory lymphohistiocytic infiltration with admixture of neutrophils in the mucosa and stroma renal pelvis. The revealed changes indicate the development of the animal acute pyelonephritis.

The positive effect of using the proposed method consists in preventing pyelonephritis in experimental animals on the background of violations of the composition of the intestinal microflora, resulting from improper feeding of animals.

Method of prevention of pyelonephritis in the experiment, which consists in the fact that the experimental animals with impaired intestinal microflora composition, within 10 days to give the drug, "Ultrasorb" from the calc is 3 g/day, mixing it with food.



 

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1 tbl, 1 ex, 6 ex

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3 tbl, 2 ex

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4 ex

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6 tbl, 3 dwg, 3 ex

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