Experimental model of acute urate oxalate nephrolithiasis in Wistar rats

Введение. В последнее время возросла частота регистрации случаев уратно-оксалатного нефролитиаза. Для разработки новых методов фармакологической коррекции и оценки эффективности фармакотерапии необходимо создание адекватной модели заболевания. Цель исследования - моделирование смешанного уратно-оксалатного нефролитиаза на крысах Wistar и изучение биохимической и гистологической картины патологии. Методика. Эксперимент выполнен на 30 самцах крыс Wistar, разделенных на группу сравнения (n=10) и экспериментальную группу (n=20). Животным экспериментальной группы ежедневно на протяжении 7 сут внутрижелудочно через зонд вводили смесь оксониевой и мочевой кислот в дозах 500 и 1000 мг/кг соответственно и предоставляли в качестве питья 1% раствор этиленгликоля. В моче крыс обеих групп измеряли активность лактатдегидрогеназы и γ-глутамилтрансферазы, в гомогенате почек - концентрацию тиобарбитуратреактивных продуктов, общую прооксидантную и антиоксидантную активность, активность глутатионпероксидазы, каталазы, супероксиддисмутазы. Наличие конкрементов в почках и наличие воспалительной реакции оценивали по результатам морфологического исследования. Результаты. В моче крыс экспериментальной группы к 7-м сут эксперимента активность лактатдегидрогеназы возросла более чем в 20 раз относительно исходного уровня, в группе сравнения не происходило статистически значимых изменений активности лактатдегидрогеназы по сравнению с исходным уровнем. Концентрация тиобарбитуратреактивных продуктов и активность каталазы в гомогенате почек крыс экспериментальной группы на 7-е сут исследования были в 1,3 и 1,6 раза соответственно выше аналогичных показателей группы сравнения, а общая прооксидантная и общая антиоксидантная активность, а также активность супероксиддисмутазы в гомогенате почек крыс подопытной группы статистически значимо снизились в 3,1, 1,8 и 3,2 раза соответственно относительно уровня группы сравнения. В группе сравнения почечные конкременты отсутствовали, а в экспериментальной были выявлены в 100% случаев в количестве 20,2±2,35 со средней площадью 540,55±47,7 мкм2 с признаками сопутствующих воспалительных и дистрофических процессов. Заключение. При недельном применении 1% раствора этиленгликоля и смеси оксониевой и мочевой кислот в дозах 500 и 1000 мг/кг соответственно формируется уратно-оксалатный нефролитиаз, характеризующийся повышением активности лактатдегидрогеназы в моче, активацией окислительных процессов и образованием смешанных депозитов с наличием выраженной воспалительной инфильтрации, кистозного расширения почечных канальцев, а также дистрофических изменений канальцевого эпителия. Introduction. At present, the incidence of urate-oxalate nephrolithiasis has increased. For the development of new methods of pharmacological treatment of this disease, it is necessary to create an adequate model that makes it possible to assess the effectiveness of pharmacotherapy. The aim - create a model of mixed urate-oxalate nephrolithiasis in Wistar rats and to study the biochemical and histological picture of the pathology. Methods. Experiments were carried out on 30 male Wistar rats, divided into a comparison group (n = 10) and an experimental group (n = 20). For formation of urate-oxalate nephrolithiasis, the experimental group was injected daily for 7 days intragastrically a mixture of oxonic and uric acids at doses of 500 and 1000 mg/kg, respectively, and these rats were also provided a drinking 1% solution of ethylene glycol. The activities of lactate dehydrogenase and γ-glutamyl transferase were measured in the urine of both groups. In the kidney homogenate, the concentration of thiobarbiturate-reactive products, total pro-oxidant and antioxidant activity, and activities of glutathione peroxidase, catalase, and superoxide dismutase were measured. Severity of lithogenesis, and inflammation was assessed based on the results of morphological studies. Results. In the urine of rats of the experimental group, the activity of lactate dehydrogenase increased more than 20 times relative to the initial level. In the comparison group, there were no significant biochemical changes. The concentration of thiobarbiturate-reactive substances and the activity of catalase in the kidney homogenate experimental group on the 7th day the were 1.3 and 1.6 times higher, respectively, than those of the comparison group. The total pro-oxidant and total antioxidant activity, as well as the activity of superoxide dismutase, decreased significantly by 3.1, 1.8 and 3.2 times, respectively. In the comparison group, renal calculi were absent, but in the experimental group, they were found in 100% of cases in the number of 20.2 ± 2.4 with an average area of 140.55 ± 47.7 μm2, accompanied by inflammatory and degenerative processes. Conclusion. With a weekly application of a 1% solution of ethylene glycol and a mixture of oxonic and uric acids at doses of 500 and 1000 mg/kg, respectively, a pronounced urate-oxalate nephrolithiasis is formed. This was characterized by an increase in the activity of lactate dehydrogenase in the urine, activation of oxidative processes, and the formation of mixed deposits associated with pronounced inflammatory infiltration, cystic dilatation of the renal tubules, as well as dystrophic changes in the tubular epithelium.

Effect of Seed Extracts of Vigna unguiculta and Hordeum vulgare on Kidney Homogenate and Rat Kidney Injury Molecule-1 in Ethylene Glycol and Ammonium Chloride Induced Urolithasis in Rats

The experiment was carried out to study the therapeutic efficacy of aqueous, alcoholic, and biherbal extracts of Vigna unguiculata (VU) and Hordeum vulgare (HV) in ethylene glycol and ammonium chloride-induced urolithiasis in female Wistar rats. Rats were divided into 14 groups, each of 6 rats, except the lithiatic control group, which consisted of 8 rats. Group I and II served as lithiatic and vehicle control, respectively. In group I and III to XIV urolithiasis were induced by administration of 0.75 % (v/v) ethylene glycol and 2% (w/v) ammonium chloride along with drinking water for 28 days. Group II was given 0.5% sodium bicarbonate. After the 28th day, the rats of urolithiatic treatment Groups III to XIV were given aqueous and alcoholic seed extracts of VU and HV @ 200 mg/kg and 300 mg/kg b.wt. orally as either single extract or combination as biherbal extracts (1:1) in 0.5 % sodium bicarbonate for another 35 days using syringe and rat gavage needle. Blood samples were collected twice: on the 28th day of induction of urolithiasis and 63rd day experiment/herbal treatment. Significantly (p less than 0.01) increased levels of calcium, oxalate, phosphate, and decreased levels of magnesium in the kidney homogenate were observed in the calculi induced groups as compared to the vehicle control group on 28th day. However, significantly increased rat Kidney Injury Molecule-1 (KIM-1) was observed in the calculi induced groups as compared to the vehicle control group in serum on the 28th day. Results of kidney homogenate and KIM-1 revealed that aqueous and alcoholic extracts of VU and HV possess good therapeutic efficacy against urolithiasis. The effect of biherbal alcoholic extract of the seeds at higher dose rate was much better in reducing or normalizing the values of most traits by 35 days of treatment, i.e., by 63rd day of the experiment and thus the continuation of treatment for some more days would be expected to restore the normal profile.

Anti-urolithiatic Activity of Sambong (Blumea balsamifera) Extract in Ethylene Glycol-induced Urolithiatic Wistar Rats (Rattus norvegicus)

Objective. The study aimed to determine if Blumea balsamifera inhibits calcium oxalate stone formation in the kidneys through determination of the number of calcium oxalate stones in the renal cortex and the percent mass of calcium oxalate. Methods. Post-test only control group design was used using five treatment groups with placebo as the negative control, potassium citrate as the positive control, and 50%, 100%, and 200% sambong treatment. Urolithiasis was induced through ethylene glycol and ammonium chloride. Each treatment group was administered its corresponding treatment solution once daily for twenty-one days. Histopathologic examination and kidney homogenate analysis were done to determine the degree of deposition of calcium oxalate stones in renal tissues and the oxalate content, respectively. Statistical analyses were performed using one-way ANOVA and post hoc Gabriel's Pairwise Comparisons Test. Results. The 100% sambong treatment group showed the least mean number of stones while the positive control and 50% sambong treatment group exhibited the highest anti-urolithiatic activity in terms of oxalate content of the kidney homogenate. Conclusion. It can be concluded from the study that Blumea balsamifera inhibits calcium oxalate stone formation in the kidneys with the 100% and 50% sambong treatment most effective in decreasing number of stones and oxalate content of the kidney homogenate, respectively.

Antiurolithiatic Activity of Pisonia Alba Leaves in Experimental Animal Model (Albino Rats)

Pisoniaalbais a medicinal plant used widely has a broad variety of pharmacological uses. The current research explored the acute toxicity of an ethanol sample of Pisoniaalba (Family: Nyctaginaceae) Ethanol Leaves Extract Pisoniaalba(ELPA) and Pisoniaalba(ALPA) Aqueous Leaves Extract at a dose of 1000mg / kg and comparison of urinary amount and electrolyte amount as parameters of evaluation. On the other side, urolithiasis caused by ethylene glycol (0.75% v / v in drinking water for 28 days) was used for the same ELPA and ALPA with an initial dose of 250 mg / kg in male rats for antiurolithiatic impact. As a conventional comparison drug, cystone (250 mg / kg, p.o.) was used amount of different urolithiatic parameters in the biological samples i.e (urine, serum and kidney homogenate) and renal function were used as criteria for evaluating the antiurolithiatic impact of ELPA and ALPA after 28days of therapy span results indicated that, in a model similar to that of furosemide, the ELPA and ALPA (250mg / kg) elevate blood quantity and urinary excretion of electrolytes. ELPA and ALPA reduced the excretion and deposition of multiple urolithiatic parameters in ethylene glycol-induced urolithiatic model relative to urolithiatic command in a pattern similar to cystone control. Supplementation with ELPA and ALPA also avoids renal function deficiency. According to our studies, ELPA supports stronger anti-urolithic exercise.

EVALUATION OF ANTINEPHROLITHIATIC ACTIVITY OF MALLOTUS PHILIPPINENSIS LINN. IN RATS

The present study was aimed to evaluate antinephrolithiatic activity of alcoholic extract of Mallotus philippinensis Linn. on ethylene glycol (0.75% V/V) induced nephrolithiasis in rat model. The leaves of Mallotus philippinensis Linn. were extracted and antinephrolithiatic activity was evaluated at the dose of 250 & 500 mg/kg b.w., p.o. Treatment with alcoholic leaves extract of Mallotus philippinensis decreased levels of calcium oxalate, phosphate in urine and kidney homogenate. Serum levels of creatinine, urea and uric acid were also decreased significantly in curative as well as protective group as compared to model control rats. The alcoholic extract of Mallotus philippinensis Linn. at the dose of 500 mg/kg showed no microcrystalline deposition and kidney damage which enables us to confirm the protective and curative potential of Mallotus philippinensis Linn. on ethylene glycol induced nephrolithiasis in rats. It can be concluded that alcoholic leaves extract of Mallotus philippinensis possess a significant antinephrolithiatic activity.

Long-term aldosterone administration increases renal Na+-Cl− cotransporter abundance in late distal convoluted tubule

Renal Na+-Cl− cotransporter (NCC) is expressed in early distal convoluted tubule (DCT) 1 and late DCT (DCT2). NCC activity can be stimulated by aldosterone administration, and the mechanism is assumed to depend on the enzyme 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2), which inactivates glucocorticoids that would otherwise occupy aldosterone receptors. Because 11β-HSD2 in rat may only be abundantly expressed in DCT2 cells and not in DCT1 cells, it has been speculated that aldosterone specifically stimulates NCC activity in DCT2 cells. In mice, however, it is debated if 11β-HSD2 is expressed in DCT2 cells. The present study examined whether aldosterone administration in mice stimulates NCC abundance and phosphorylation in DCT2 cells but not in DCT1 cells. B6/C57 male mice were administered 100 µg aldosterone·kg body weight−1·24 h−1 for 6 days and euthanized during isoflurane inhalation. Western blotting of whole kidney homogenate showed that aldosterone administration stimulated NCC and pT58-NCC abundances ( P < 0.001). In DCT1 cells, confocal microscopy detected no effect of the aldosterone administration on NCC and pT58-NCC abundances. By contrast, NCC and pT58-NCC abundances were stimulated by aldosterone administration in the middle of DCT2 ( P < 0.001 and <0.01, respectively) and at the junction between DCT2 and CNT ( P < 0.001 and <0.05, respectively). In contrast to rat, immunohistochemistry in mouse showed no/very weak 11β-HSD2 expression in DCT2 cells. Collectively, long-term aldosterone administration stimulates mouse NCC and pT58-NCC abundances in DCT2 cells and presumably not in DCT1 cells.

Evaluation of Antiurolithiatic Activity of Lawsonia inermis Linn. in Rats

This study was conducted to evaluate the potential antiurolithiatic effects of Lawsonia Inermis Linn. in rat models. Animals were divided into seven groups and urolithiasis was induced by ethylene glycol (0.75% v/v) in drinking water to all groups (Groups II-VII) except normal control (Group I) for 28 days. Methanolic extracts of Lawsonia inermis (MELI) bark (300 & 500 mg/kg, p.o.) were administered once daily, from 15th day to 28th day as curative regimen and from 1st day to 28th day as preventive regimen. Cystone (750 mg/kg, p.o.) was used as a standard drug for comparison. After 28 days, various biochemical parameters like urine volume, pH were measured. Calcium, phosphate and oxalate were measured in urine and kidney homogenate. Serum creatinine, uric acid and urea nitrogen were estimated. Histopathology of kidney was investigated. Treatment with the MELI extract significantly (p<0.05) restored all elevated parameters including calcium, phosphate and oxalate in urine and kidney homogenate; creatinine, uric acid and urea nitrogen in serum when compared to model control group. The histopathological study of the kidney also supported the above results. It was concluded that methanolic extract of bark of Lawsonia inermis Linn. has significant antiurolithiatic effect in experimental rats.

Evaluation of Antiurolithiatic Activity of Lawsonia inermis Linn. in Rats

Lawsonia inermis Linn., commonly called as Henna belongs to family Lythraceae. Traditionally, it has been reported to have therapeutic activity in bronchitis, diabetes, antimicrobial, antibacterial, trypsin inhibitory, cytotoxicity, wound healing, antioxidant, anti-inflammatory, analgesic. However, the antiurolithiatic activity of the bark extract of L. inermis Linn. is not known. In this study, we investigated protective effect of the alcoholic extract of L. inermis bark against ethylene glycol induced urolithiasis and its possible underlying mechanisms using male wistar albino rats. Animals were divided into seven groups and urolithiasis was induced by ethylene glycol (0.75% v/v) in drinking water for 28 days. Methanolic extract of Lawsonia inermis (MELI) bark (300 & 500 mg/kg, p.o.) were administered once daily from 15th day to 28th day as curative regimen and from 1st day to 28th day as preventive regimen. Cystone (750 mg/kg, p.o.) was used as a standard drug. After 28 days, various biochemical parameters like urine volume, pH were measured. Calcium, phosphate and oxalate were measured in urine and kidney homogenate. Serum creatinine, uric acid and urea nitrogen were estimated. Histopathology of kidney also studied. Treatment with the MELI extract significantly restored all elevated parameters including calcium, phosphate and oxalate in urine and kidney homogenate; creatinine, uric acid and urea nitrogen in serum when compared to model control group. The histopathological study of the kidney also supported the above results. It can be concluded that methanolic extract of bark of Lawsonia inermis Linn. has significant antiurolithiatic effect in experimental rats.

NTA and Fe(III)NTA: differential patterns of renal toxicity in subchronic studies

Differential patterns in terms of nephropathology and 8-hydroxyguanine formation in the course of oral 28-day studies were observed with nitrilotriacetic acid (NTA) and FeNTA. FeNTA, but not NTA, caused enhanced 8-hydroxyguanine formation in kidney DNA after oral and intraperitoneal administration. Enhanced lipid peroxidation in the kidney homogenate was observed with FeNTA as well as with NTA. For NTA, the low dose (9 mg/kg per day) was without adverse effect. The kidney toxicity of oral FeNTA (50, 200, and 1000 mg/ kg per day) was only mild, 50 mg/ kg per day; however, it still led to an increased 8-hydroxyguanine content. The relevance of Iron(III) (Fe(III)) or Fe(III)NTA formation as a relevant mediator of NTA-related toxicity was excluded on the basis of these data. Also, a thermodynamic consideration presented here, supports the view that zinc (Zn), and not Fe, is likely to mediate the tubular cell cytotoxicity of NTA.