Administration Of Ammonium Chloride Research Articles

Ammonium chloride-induced hypothermia is attenuated by transient receptor potential channel vanilloid-1, but augmented by ankyrin-1 in rodents

AimsSystemic administration of ammonium chloride (NH4Cl), an acidifying agent used in human patients and experimental conditions, causes hypothermia in mice, however, the mechanisms of the thermoregulatory response to NH4Cl and whether it develops in other species remained unknown. Main methodsWe studied body temperature (Tb) changes in rats and mice induced by intraperitoneal administration of NH4Cl after blockade of transient receptor potential vanilloid-1 (TRPV1) or ankyrin-1 (TRPA1) channels. Key findingsIn rats, NH4Cl decreased Tb by 0.4–0.8°C (p < 0.05). The NH4Cl-induced hypothermia also developed in Trpv1 knockout (Trpv1−/−) and wild-type (Trpv1+/+) mice, however, the Tb drop was exaggerated in Trpv1−/− mice compared to Trpv1+/+ controls with maximal decreases of 4.0 vs. 2.1°C, respectively (p < 0.05). Pharmacological blockade of TRPV1 channels with AMG 517 augmented the hypothermic response to NH4Cl in genetically unmodified mice and rats (p < 0.05 for both). In contrast, when NH4Cl was infused to mice genetically lacking the TRPA1 channel, the hypothermic response was significantly attenuated compared to wild-type controls with maximal mean Tb difference of 1.0°C between the genotypes (p = 0.008). Pretreatment of rats with a TRPA1 antagonist (A967079) also attenuated the NH4Cl-induced Tb drop with a maximal difference of 0.7°C between the pretreatment groups (p = 0.003). SignificanceTRPV1 channels limit, whereas TRPA1 channels exaggerate the development of NH4Cl-induced hypothermia in rats and mice, but other mechanisms are also involved. Our results warrant for regular Tb control and careful consideration of NH4Cl treatment in patients with TRPA1 and TRPV1 channel dysfunctions.

Ammonium chloride administration prevents training-induced improvements in mitochondrial respiratory function in the soleus muscle of male rats.

Exercise training can increase both mitochondrial content and mitochondrial respiration. Despite its popularity, high-intensity exercise can be accompanied by mild acidosis (also present in certain pathological states), which may limit exercise-induced adaptations to skeletal muscle mitochondria. The aim of this study was to determine if administration of ammonium chloride (0.05 g/kg) to Wistar rats before each individual exercise session (5 high-intensity exercise sessions per week for eight weeks) reduced training-induced increases in mitochondrial content (measured by citrate synthase activity and protein content of electron transport system complexes) and respiration (measured in permeabilised muscle fibres). In the soleus muscle, the exercise-training-induced increase in mitochondrial respiration was reduced in rats administered ammonium chloride compared to control animals, but mitochondrial content was not altered. These effects were not present in the white gastrocnemius muscle. In conclusion, ammonium chloride administration before each exercise session over eight weeks reduced improvements in mitochondrial respiration in the soleus muscle but did not alter mitochondrial content. This suggests that mild acidosis may impact training-induced improvements in the respiration of mitochondria in some muscles.

Pharmacological Evaluation of Mentha piperita Against Urolithiasis: An In Vitro and In Vivo Study.

Background: Mentha piperita L. (peppermint) is one of the most widely consumed medicinal herbs that has gained attention from food and pharmaceutical industries due to its distinct aroma and taste. Purpose: Present study was aimed to rationalize the traditional use of peppermint in urolithiasis and to explore its possible underlying mechanism. Research Design: The aqueous methanolic crude extract of Mentha piperita (Mp.Cr) was assessed for phytochemical constituents and antioxidant activity. In vitro crystallization assays were performed to determine the inhibitory effects of Mp.Cr against crystal nucleation, aggregation and growth. In vivo urolithiasis model was developed in rats by the administration of ammonium chloride and ethylene glycol in drinking water. The antiurolithic effects of Mp.Cr were evaluated by analyzing kidney homogenate, biochemical and histological parameters. Results: HPLC analysis showed the presence of epicatechin, quercetin, gallic acid, syringic acid, kaempferol, caffeic acid and coumaric acid. The maximum quantity of quercetin equivalent flavonoid and gallic acid equivalent phenolic content was found to be 63.73 ± .24 mg QE/g and 43.76 ± .6 mg GAE/g of Mp.Cr, respectively. Mp.Cr significantly normalized urinary and serum biochemistry, similar to the standard cystone treatment. Conclusions: The current study validated the preventive and curative potential of Mp.Cr against urolithiasis and justified its traditional use in kidney stone disease.

Use of ammonium chloride to prevent urolithiasis in sheep

ABSTRACT Urolithiasis has a high incidence among confined sheep. It is multifactorial and may cause economic damage. Our aim was to determine the capacity of urinary acidification using ammonium chloride in sheep. Twenty-five 3-month-old male sheep were confined and randomly divided into three groups; the G200 and G500 groups received 200mg/kg/GW and 500mg/kg/GW of ammonium chloride daily for 56 consecutive days, respectively, whereas the CG group did not receive ammonium chloride. Sampling times and clinical evaluation were performed weekly, starting from the 14th day of confinement (M1 or immediately before administering ammonium chloride) until the 17th day (M9) of the feedlot. Hemogasometry, biochemical examination of serum urea and creatinine concentration and ultrasound evaluation of the urinary tract were performed. The urinalysis indicated a higher incidence of ammonium magnesium phosphate crystals at the beginning of the study, showing a migration to urate crystal formation, mainly in the G500 group because of urinary acidification. No hemogasometric, serum biochemistry, ruminal fluid, or ultrasonographic changes were observed. Urinary acidification was achieved and maintained after M7 during the administration of ammonium chloride in the G500 group, but not in the other study groups.

Ultrasonographic application in the diagnosis and treatment of cystitis‐ pyelonephritis syndrome in buffaloes (Bubalus bubalis): A novel approach

AbstractBovine cystitis is an inflammation of the urinary bladder that may ascend to ureters to cause pyelonephritis. The aim of this study performed in buffaloes was to investigate the use of ultrasonographic imaging in the diagnosis of cystitis‐pyelonephritis syndrome followed by a novel treatment regimen. In all four buffaloes, diagnosis was confirmed using trans‐rectal ultrasonography, revealing non‐specific thickening of bladder wall, with reduced capacity and floating hyperechoic contents. Kidneys showed dilatation of right and/or left ureter, with loss of lobulations and cortex medullary differentiation. Successful treatment was achieved by flushing of the urinary bladder using a number 18 Foleys catheter guided by ultrasound twice a day for 5 days with 2.5% liquid betadine in combination with oral administration of ammonium chloride (0.2 mg/kg body weight) for 7 days. A rapid recovery was confirmed by clinical examination after 7 days and a further follow‐up 2 months post‐treatment.

Ammonium Salts Promote Adaptive Changes of Rat Immune System to Ultimate Load in the Forced Swimming Model.

We studied the effect of different doses of ammonium chloride (ACl) and ammonium carbonate (ACr) on immunological parameters of the peripheral blood in rats during high-intensity exercise. Changes in the absolute and relative numbers of granulocytes, lymphocytes, natural killers, naive and mature effector cells one day after the end of the forced swimming cycle were found by using a hematological analyzer and a flow cytometer. Immunological indicators were analyzed relative to swimming duration on the last day of ultimate load. The revealed changes indicate the onset of the effector phase of the development of the inflammatory processes in the positive control group (physiological saline) and in rats receiving a higher dose of ACr (20 mg/kg), while administration of ACl prevented the development of inflammatory processes and shifts in the physiological balance of lymphocyte subpopulations. Immunological profiling indicates that ACl in a dose of 20 mg/kg most effectively improved physical performance in our forced swimming model.

6-Shogaol attenuated ethylene glycol and aluminium chloride induced urolithiasis and renal injuries in rodents.

The 6-shogaol, is a flavanone type flavonoid that is abundant in citrus fruit and has a wide range of pharmacological effects. The present study attempted to evaluate the antiurolithic effect of 6-shogaol on ethylene glycol (EG) and ammonium chloride (AC)-induced experimental urolithiasis in rats. The efficacy of 6-shogaol 50 mg/kg and 100 mg/kg was studied in EG 0.75% (V/V) and AC 1% (W/V) experimentally induced urolithiasis in rats for 21 days. The weight difference, urine volume, the levels of calcium, phosphate, magnesium, oxalate and uric acid in urine was observed. The blood urea nitrogen, creatinine, uric acid in serum and levels of malondialdehyde (MDA) and glutathione (GSH) were also measured. Histopathological analyses in kidneys were also performed. The rats weights were higher in the 6-shogaol groups than the urolithiasis group. EG caused a significant increase in serum creatinine (p < 0.05), BUN (P < 0.001), and uric acid (p < 0.01) while treatment with Cystone (750 mg/kg), and 6-shogaol (50 and 100 mg/kg) showed the significant reduction in increased serum levels of creatinine (p < 0.001), uric acid (p < 0.01) and BUN (p < 0.001). Administration of EG and AC showed statistically significant (p < 0.001) elevated levels of MDA and reduction in GSH levels. Treatment of Cystone (750 mg/kg), and 6-shogaol (50 and 100 mg/kg) significantly (p < 0.001) reduced MDA levels and an increase GSH levels as compared to EG and AC-treated group. The histological findings further attested antiurolithiatic properties of 6-shogaol. The present study attributed clinical shreds of evidence first time that claiming the significant antiurolithic effect of 6-shogaol and could be a cost-effective candidate for the prevention and treatment of urolithiasis.

Ammonium chloride administration prior to exercise has muscle-specific effects on mitochondrial and myofibrillar protein synthesis in rats.

AimExercise is able to increase both muscle protein synthesis and mitochondrial biogenesis. However, acidosis, which can occur in pathological states as well as during high‐intensity exercise, can decrease mitochondrial function, whilst its impact on muscle protein synthesis is disputed. Thus, the aim of this study was to determine the effect of a mild physiological decrease in pH, by administration of ammonium chloride, on myofibrillar and mitochondrial protein synthesis, as well as associated molecular signaling events.MethodsMale Wistar rats were given either a placebo or ammonium chloride prior to a short interval training session. Rats were killed before exercise, immediately after exercise, or 3 h after exercise.ResultsMyofibrillar (p = 0.036) fractional protein synthesis rates was increased immediately after exercise in the soleus muscle of the placebo group, but this effect was absent in the ammonium chloride group. However, in the gastrocnemius muscle NH4Cl increased myofibrillar (p = 0.044) and mitochondrial protein synthesis (0 h after exercise p = 0.01; 3 h after exercise p = 0.003). This was accompanied by some small differences in protein phosphorylation and mRNA expression.ConclusionThis study found ammonium chloride administration immediately prior to a single session of exercise in rats had differing effects on mitochondrial and myofibrillar protein synthesis rates in soleus (type I) and gastrocnemius (type II) muscle in rats.

Ammonium Salts Promote Functional Adaptation of Rat Erythrocytes on the Model of Forced Swimming

Ammonium, an end-product of catabolism, in low doses can promote adaptation of metabolic pathways in erythrocytes under conditions of extreme physical exercise. We compared the effects of two ammonium salts, ammonium chloride and ammonium carbonate, in two doses on biochemical parameters of rat erythrocytes 1 day after extreme physical exercise in a 4-week cycle of forced swimming. Of 16 analyzed parameters, the maximum number of significant shifts from the control was revealed in the groups of rats receiving ammonium chloride in doses of 20 and 10 mg/kg, and the minimal number of differences was found in groups treated with ammonium carbonate in the same doses. The comparison of the levels of reduced glutathione and 2.3-bisphosphoglicerate and activities of 5'-nucleotidase and Ca2+- and Na/K-ATPases attested to more rigorous control of the mechanism of oxygen delivery to tissues by erythrocytes after administration of ammonium chloride in a dose of 20 mg/kg.

Anti-urolithiatic activity of Solanum nigrum hydroalcoholic extract in ethylene glycol-induced urolithiasis in rats

Background and objectives Urolithiasis is a growing public health problem. Asymptomatic kidney stones are kept under observation. The aim of this study was to explore the anti-urolithiatic activity of the hydroalcoholic extract of Solanum nigrum fruit in ethylene glycol-induced urolithiasis in rats. Materials and methods Urolithiasis was induced by oral administration of ammonium chloride 1% and ethylene glycol (0.75% v/v) in drinking water for 28 days. Hydroalcoholic extract of Solanum nigrum fruit (200 and 400 mg/kg) and cystone (750 mg/kg) were administered orally from the 15th day as a curative regimen. Results and conclusion Administration of ethylene glycol caused an elevation of serum creatinine, urea, calcium, and malondialdehyde and a reduction of magnesium and glutathione. In addition, renal content of tumor necrosis factor alpha was elevated and adiponectin renal content was reduced in urolithiatic control. Histopathological examination revealed tubular degeneration, dilatation, presence of calcium oxalate crystals in the lumen of renal tubules, and intense interstitial mononuclear cell infiltration in the lithiatic control group. Treatment with both doses of Solanum nigrum reversed all biochemical parameters and histopathological alterations. The results demonstrate that the hydroalcoholic extract of Solanum nigrum has potent anti-urolithiatic activity against calcium oxalate urolithiasis induced by ethylene glycol through tumor necrosis factor alpha inhibition and adiponectin stimulation as well as in maintaining balance between stone promoter (calcium) and inhibitor (magnesium).

ВИЗНАЧЕННЯ ВІДХАРКУВАЛЬНОЇ АКТИВНОСТІ ТАБЛЕТОК НА ОСНОВІ ЕКСТРАКТІВ ІЗ КВІТОК МАЛЬВИ ЛІСОВОЇ ТА ПОДОРОЖНИКА ЛАНЦЕТОЛИСТОГО

Вступ. Згідно з останніми даними ВООЗ, сьогодні захворювання дихальної системи є провідною причиною смерті та інвалідності у світі. Відповідно до формулярної системи України (2009), лікування гострих бронхолегеневих захворювань інфекційного походження починають з етіотропних засобів, до яких приєднують патогенетичні монопрепарати. До цієї групи належать препарати рослинного походження і препарати резорбтивної дії.&#x0D; Мета дослідження – вивчити відхаркувальні властивості таблеток, до складу яких входять екстракти з квіток мальви лісової та подорожника ланцетолистого.&#x0D; Методи дослідження. Для експерименту було використано здорових мишей-самців BALB/c (20–25 г). Через 10 днів адаптації тварин випадковим чином поділили на групи по 6 мишей у кожній. Усім тваринам речовини вводили атравматичним зондом інтрагастрально: мишам 1-ї групи – воду дистильовану; 2-ї – розчин амонію хлориду (1,0 г/кг маси тіла тварини); 3–5 – розчинені екстракти з квіток мальви лісової (0,062 г/кг), подорожника ланцетолистого (0,062 г/кг) та досліджувані таблетки (0,4819 г/кг). Через 30 хв після останнього введення мишам вводили внутрішньочеревно розчин фенолового червоного (5 % у фізіо­логічному розчині, 0,1 мл/10 г маси тіла). Після впливу ультразвуку на ізольовані органи протягом 10 хв додавали 0,1 мл 1 М розчину натрію гідроксиду. Для вимірювання оптичної щільності одержаного розчину при довжині хвилі 546 нм було використано спектрофотометр Lambda 25 (“PerkinElmer”, США). &#x0D; Результати й обговорення. Введення експериментальним тваринам амонію хлориду, екстрактів із квіток мальви лісової та подорожника ланцетолистого, а також таблеток стимулювало секреторну функцію бронхів. Дана експериментальна модель дала змогу охарактеризувати відхаркувальні властивос­ті досліджуваного препарату та визначити його вплив на секреторну функцію бронхів: за відхаркувальною здатністю досліджуваний засіб на 40,16 % перевищував препарат порівняння (амонію хлорид).&#x0D; Висновок. Досліджувані таблетки мають високу здатність секретувати мокротиння, проявляють хорошу терапевтичну активність та можуть бути рекомендовані для подальших досліджень як потенційний препарат із відхаркувальними властивостями.

Effect of urine pH and magnesium on calcium oxalate saturation.

Hypomagnesiuria is a common biochemical finding in patients with calcium oxalate (CaOx) nephrolithiasis. Clinical trials using Mg supplements as therapy against CaOx stones have shown mixed results. We tested the effect of Mg administration in healthy subjects under conditions of controlled urine pH (UpH) on urinary Ca excretion rate (UCaV) and CaOx saturation. This is a 4-phase, double blind, placebo-controlled, metabolic crossover study performed in healthy volunteers. Mg lactate (MgLact2) was used as Mg supplement. High UpH and low UpH were achieved by administration of potassium citrate (K3Citrate) and ammonium chloride (NH4Cl), respectively, with potassium balance maintained by KCl. Eight participants completed 4 phases of study. The interventions successfully modulated 24-h UpH (7.0±0.4 vs. 5.7±0.6 in high vs low pH phases; P<0.001). Administration of MgLact2 increased UMgV [175.8±40.2 vs 93.4±39.7mg/day (7.2±1.7 vs 3.8±1.6mmol/day), high vs low Mg phase; P<0.001], and increased pH both at low (5.6±0.5 to 5.8±0.7; P=0.02) and high UpH (6.9±0.4 to 7.0±0.3; P=0.01). At a given urine pH, Mg supplementation marginally increased UCaV, but did not alter UOxV or CaOx saturation. Provision of an alkali load significantly lowered UCaV and saturation of CaOx at any level of UMgV. Compared to changes in UMgV, changes in UpH play a more significant role in determining urine CaOx saturation in healthy subjects. Mg supplements are likely to reduce CaOx saturation if they also raise urine pH.

Alteration in Oxidative/nitrosative imbalance, histochemical expression of osteopontin and antiurolithiatic efficacy of Xanthium strumarium (L.) in ethylene glycol induced urolithiasis.

Xanthium strumarium has traditionally been used in the treatment of urolitiasis especially by the rural people in India, but its antiurolithiatic efficacy was not explored scientifically till now. Therefore, the present study was designed to validate the ethnic practice scientifically, and explore the possible antiurolithiatic effect to rationalize its medicinal use. Urolitiasis was induced in hyperoxaluric rat model by giving 0.75% ethylene glycol (EG) for 28days along with 1% ammonium chloride (AC) for first 14days. Antiurolithiatic effect of aqueous-ethanol extract of Xanthium strumarium bur (xanthium) was evaluated based on urine and serum biochemistry, oxidative/nitrosative stress indices, histopathology, kidney calcium and calcium oxalate content and immunohistochemical expression of matrix glycoprotein, osteopontin (OPN). Administration of EG and AC resulted in hyperoxaluria, crystalluria, hypocalciuria, polyurea, raised serum urea, creatinine, erythrocytic lipid peroxidise and nitric oxide, kidney calcium content as well as crystal deposition in kidney section in lithiatic group rats. However, xanthium treatment significantly restored the impairment in above kidney function test as that of standard treatment, cystone. The up-regulation of OPN was also significantly decreased after xanthium treatment. The present findings demonstrate the curative efficacy of xanthium in ethylene glycol induced urolithiasis, possibly mediated through inhibition of various pathways involved in renal calcium oxalate formation, antioxidant property and down regulation of matrix glycoprotein, OPN. Therefore, future studies may be established to evaluate its efficacy and safety for clinical use.

Is calcitonin an active hormone in the onset and prevention of hypocalcemia in dairy cattle?

The objective of this study was to assess the potential importance of calcitonin (CALC) in the onset of subclinical hypocalcemia (experiment 1) and in the physiological mechanisms underlying the prevention of bovine hypocalcemia under metabolic acidosis (experiments 2 and 3). In experiment 1, 15 Holstein cows naturally incurring subclinical hypocalcemia during the first 5d postpartum were classified as low subclinical hypocalcemia (LSH) when blood Ca concentrations were between 7.5 and 8.5mg/dL, or as high subclinical hypocalcemia (HSH) when blood Ca concentrations were between 6.0 and 7.6mg/dL. Blood samples were taken daily from d −5 to 5 relative to parturition to determine concentrations of parathyroid hormone (PTH), CALC, and 1,25(OH)2D3. In experiment 2, 24 Holstein bulls (497±69kg of body weight and 342±10.5d of age) were assigned to 2 treatments (metabolic acidosis or control). Metabolic acidosis was induced by an oral administration of ammonium chloride (2.5mEq/d) during 10 d, and animals were slaughtered thereafter. Blood samples were collected before slaughter to determine CALC, PTH, 1,25(OH)2D3, and samples of urine, kidney, parathyroid, and thyroid glands were obtained immediately after slaughter to determine expression of several genes in these tissues. Last, in experiment 3, we tested the activity of CALC under metabolic acidosis in vitro using breast cancer cell (T47D) cultures. Although PTH tended to be greater in HSH than in LSH, the levels of 1,25(OH)2D3 were lower in HSH cows (experiment 1). Blood CALC concentration was not affected by the severity of subclinical hypocalcemia, but it was influenced by days from calving (experiment 1). The expression of PTH receptor (PTHR) in the kidney was increased under metabolic acidosis (experiment 2). Furthermore, the activity of CALC was impaired under acidic blood pH (experiment 3). In conclusion, the CALC rise in HSH cows after calving impaired the recovery of blood Ca concentrations because the PTHR response was not sufficient to activate 1,25(OH)2D3 and compensate for the CALC effect. Metabolic acidosis prevents hypocalcemia because the expression of PTHR is upregulated in the kidney, resulting in an increased PTH activity and a subsequent increase in 1,25(OH)2D3 serum concentrations. Moreover, an impairment of CALC activity at low pH enhances the hypercalcemic role of PTH.

Surgical management of urinary obstruction in young ruminants by tube cystotomy: A report of 24 cases

Objectives: To assess the efficacy of tube cystotomy along with medical dissolution of calaculi as a sole strategy for correction of obstructive urolithiasis in young ruminants. Materials and methods: Young bull calves (n=24; 10 Ongole claves and 14 Murrah buffalo calves) suffering from complete/partial urinary retention aging between 1 to 6 months were treated by tube cystotomy along with oral administration of ammonium chloride dosed at 10 gm/Kg body weight (bwt) orally for 15 days, streptopencillin dosed at 100 mg/10 Kg bwt through intramuscular (im) route for 5 days, and meloxicam dosed at 0.2 mg/Kg bwt through im route for 3 days. Results: Total 23 (95.83%) out of 24 calves started passing urine normally through the natural orifice 10-15 days postoperatively. None of the recovered animals (n=23) exhibited recurrence of symptoms, establishing the superiority of the technique in resolving the condition. Conclusion: Tube cystotomy, when performed at an early stage can prevent mortality of calves due to cystorrhexis, uroperitoneum and consequent uremia. This can avoid the painful and most tedious cysto-urethrotomy in young ruminants. http://doi.org/10.5455/javar.2016.c142

The urinary bladder carcinogen propoxur does not produce genotoxic effects in the urinary bladder of Wistar male rats

Propoxur (PPX) is a carbamate insecticide which induced urinary bladder cancer in Wistar rats when fed at 5000ppm in Altromin 1321 diet (1321). In the present investigation, PPX was studied for induction of several key events related to modes of action (MOA) of carcinogenicity in urinary bladders (UBs). Wistar rats were administered the compound for 28 days at 8000ppm in Provini Liba SA 3883 diet, which is similar to the 1321 diet. o-Anisidine HCl (AH) was used as a genotoxic UB carcinogenic comparator, and trisodium nitrilotriacetate (NTA) as an epigenetic UB carcinogen comparator. Along with the non-dosed control and three test substance groups (PPX, AH, NTA), four more groups were additionally fed 2% ammonium chloride (AC) in the diet to acidify the urine, since 1321 was reported to increase urinary pH. AC did acidify the urine, as expected, although the 3883 diet itself did not increase pH values above 8. In the alkaline comet assay, AH produced DNA single strand breaks (SSBs) in the UB urothelium (UBU) irrespective of AC administration, whereas PPX and NTA did not. In the nucleotide 32P-postlabeling assay (NPL), AH produced DNA adducts irrespective of AC administration, whereas PPX and NTA did not. Routine (H&E) histopathology evaluation of the UBU did not reveal any hyperplasia or evidence of luminal microprecipitates or calculi in any of the groups. Assessment of UBU proliferation as measured by immunohistochemistry of proliferating cell nuclear antigen, revealed that NTA and NTA plus AC increased the replicating fraction (RF). Also AH plus AC, but not AH alone, increased the RF of UBU, whereas PPX groups were not significantly different from controls. Thus, the results reveal no evidence for DNA SSBs, binding, or alteration of DNA synthesis in the UBU by PPX, while demonstrating UBU DNA damage by AH and showing that NTA does not damage DNA, but causes increased UBU proliferation. The findings are in accord with a genotoxic MOA for AH, and an epigenetic MOA for NTA. The MOA of PPX does not involve genotoxicity and may be specific to the 1321 diet.

Study on inhibitory effect of EGCG on Calcium oxalate nephrolithiasis in rats and its related mechanism

In the study, the inhibitory effect of epigallocatechin gallate (EGCG) on Calcium oxalate nephrolithiasis and its possible mechanism were investigated. The rat Calcium oxalate nephrolithiasis model was induced through the combined oral administration of ethylene glycol and ammonium chloride, which was intervened with EGCG. Rat blood samples were collected to detect blood creatinine (Cr), blood urea nitrogen (BUN) and blood calcium. Rat urine samples were collected to observe and compare 24-hour urine volume, oxalic acid (Ox) and calcium in urine. Renal samples were collected to prepare tissue slices and observe the pathological changes in Calcium oxalate nephrolithiasis. The expression of osteopontin (OPN) in renal tissues was evaluated by Real-time PCR and Western blot. According to the results, compared with normal rats, rats in the nephrolithiasis model showed significant increases in Cr, BUN, urine Calcium, urine Ox and renal OPN expression (P < 0.05), but obvious decrease in 24-hour urine volume (P < 0.05); Compared with rats with nephrolithiasis, those processed with EGCG revealed remarkable declines in Cr, BUN, urine Calcium and urine Ox (P < 0.05), with significant rise in 24-hour urine volume (P < 0.05) in a concentration-dependent manner. Additionally, compared with the control group, nephrolithiasis rats showed significant pathological changes in Calcium oxalate calculus. After ECCG treatment, the renal pathological changes and OPN expression attenuated significantly in a concentration-dependent manner. The results showed that EGCG inhibits the formation of calcium oxalate nephrolithiasis in rats and shows a notable protective effect on renal functions.

Protective effect of Urtica dioica methanol extract against experimentally induced urinary calculi in rats.

Renal calculi formation is one of the most common urological disorders. Urinary stone disease is a common disease, which affects 10‑12% of the population in industrialized countries. In males, the highest prevalence of the disease occurs between the age of 20 and 40 years, while in females, the highest incidence of the disease occurs later. Previous studies have shown that long‑term exposure to oxalate is toxic to renal epithelial cells and results in oxidative stress. In the present study, a methanolic extract of aerial parts of Urtica dioica was screened for antiurolithiatic activity against ethylene glycol and ammonium chloride‑induced calcium oxalate renal stones in male rats. In the control rats, ethylene glycol and ammonium chloride administration was observed to cause an increase in urinary calcium, oxalate and creatinine levels, as well as an increase in renal calcium and oxalate deposition. Histopathological observations revealed calcium oxalate microcrystal deposits in the kidney sections of the rats treated with ethylene glycol and ammonium chloride, indicating the induction of lithiasis. In the test rats, treatment with the methanolic extract of Urtica dioica was found to decrease the elevated levels of urinary calcium, oxalate and creatinine, and significantly decrease the renal deposition of calcium and oxalate. Furthermore, renal histological observations revealed a significant reduction in calcium oxalate crystal deposition in the test rats. Phytochemical analysis of the Urtica dioica extract was also performed using liquid chromatography‑electrospray ionization tandem mass spectrometry and high-performance liquid chromatography with photodiode array detection, to determine the chemical composition of the extract. The eight chemical constituents identified in the extract were protocatechuic acid, salicylic acid, luteolin, gossypetin, rutin, kaempferol‑3‑O‑rutinoside, kaempferol‑3‑O‑glucoside and chlorogenic acid. In conclusion, the results of the present study suggest that Urtica dioica has strong antiurolithiatic activity and may have potential as a natural therapeutic agent for various urological disorders.

Efeito da suplementação de cloreto de amônio sobre os equilíbrios eletrolítico e ácido-básico e o pH urinário de ovinos confinados

A incidência da urolitíase obstrutiva em ovinos é elevada, principalmente em machos confinados, tanto para produção de carne, quanto reprodutores de alto valor genético. A acidificação urinária é um dos métodos para prevenção desta enfermidade e pode ser realizada de forma eficaz com a suplementação de cloreto de amônio na dieta, que pode propiciar a instalação de acidose metabólica. A hemogasometria avalia o equilíbrio ácido-básico sanguíneo de forma prática e fácil. Neste trabalho, avaliou-se o efeito do cloreto de amônio sobre o equilíbrio ácido-básico e eletrolítico de ovinos em confinamento para quantificar a acidose metabólica desenvolvida. Utilizaram-se 100 ovinos, machos, confinados, com idade aproximada de três meses. Constituíram-se três grupos experimentais: Grupo I (n=40), recebeu 400mg/kg/PV de cloreto de amônio/animal/dia por 21 dias consecutivos, momento da interrupção da administração do acidificante urinário (M3) e continuidade do acompanhamento clinico até o final do experimento (M6); Grupo II (n=40), 400mg/kg/PV de cloreto de amônio/animal/dia por 42 dias consecutivos; Grupo III (n=20), não recebeu cloreto de amônio durante todo o período do experimento. Os Momentos (M) de colheita de amostras e avaliação clínica foram estabelecidos com intervalo de sete dias, sendo M0 (imediatamente antes do início do tratamento com cloreto de amônio), M1 (sete dias após), M2, M3, M4, M5 e M6, totalizando 56 dias de confinamento. A alimentação consistiu de ração total, composta por 15% de feno triturado e 85% de concentrado, água e sal mineral ad libitum. Após adaptação de 15 dias à dieta de confinamento, colheram-se de todos os animais amostras de urina para mensuração do pH, e sangue venoso para hemogasometria, nos diferentes momentos. A acidificação urinária foi mantida enquanto houve a administração de cloreto de amônio nos grupos GI e GII. Os valores de Na+ e K+ permaneceram dentro da normalidade para a espécie. O cloreto de amônio provocou acidose metabólica hiperclorêmica compensada nos grupos GI e GII, comprovada pelos valores reduzidos de HCO3-, EB e SID, pelos valores elevados de Cl- e pelo pH normal. Concluiu-se que o cloreto de amônio apesar de provocar diminuição da reserva alcalina no organismo, não interferiu com o desenvolvimento dos animais, podendo ser empregado como agente preventivo da urolitíase obstrutiva em ovinos.

Molecular mechanisms involved in the protective effect of the chloroform extract of Selaginella lepidophylla (Hook. et Grev.) Spring in a lithiasic rat model

Urolithiasis is a multifaceted process, progressing from urine supersaturation to the formation of mature renal calculi. Calcium oxalate, the main component of kidney stones, has toxicological effects on renal epithelial cells. Some medicinal plants have shown pharmacological effects against renal lithiasis, such as Selaginella lepidophylla (Hook. et Grev) Spring, a plant empirically used in Mexico for its diuretic and antilithiasic activity. The plant was identified and ground, and a chloroform extract (CE) was obtained. Urolithiasis was induced in Wistar female rats by administration of ethylene glycol and ammonium chloride for 21days. Urolithiasis rats were treated with the CE (50mg/kg) for 21days. Osmolality, creatinine, sodium and potassium concentrations were measured in blood and urine. Glomerular filtration rate (GFR), and electrolytic and water balances were calculated. Urinary oxalic acid concentration was measured. Apoptosis, lipoperoxidation, ROS and p-amino hippuric acid were determined in cortical tissue. Urolithiasis rats showed a decrease of urinary flow, GFR, electrolytic balance, renal tubular secretion and ATP concentration and increase of urinary oxalic acid, lipoperoxidation, oxidative stress and apoptosis in cortical tissue. After treatment with the CE, urinary flow rate, GFR and renal tubular secretion levels were recovered; on the other hand, serum creatinine and urinary oxalic acid decreased on day 21. CE of Selaginella lepidophylla prevented the damage caused by lithiasic process by improving the active secretion in the proximal tubules, counteracting the ROS and lipoperoxidation effects by oxalate and decreased the OAT3 expression on kidney.