Evaluation of Phyllanthus niruri L. from Malaysia for In-vitro  Anti-Urolithiatic Properties by Different Solvent Extraction

The Phyllanthus niruri is traditionally used for curing of kidney disorders and urinary stones in Malaysia. Hence the current work was aimed to evaluate the effect of different solvents extract (n- hexane, ethyl acetate, methanol and water) of P. niruri for in vitro anti-urolithiatic properties in terms of inhibition activity on CaOx by using the rate of CaOx aggregation assay and dissolution of calcium oxalate (CaOx) crystal by using titrimetry method. Cystone was used as positive control. The effects of cystone on slope of nucleation and aggregation as well as growth of CaOx were evaluated spectrophotometrically. The highest yield percentage of P.niruri was occupied by methanol (5.74 %). The maximum inhibition against aggregation of CaOx crystals was also occupied by methanol (66.67 % ± 1.61) and was comprised with alkaloid, steroid, terpenoid and tannin. Dissolution effect on calcium oxalate crystals indicates that the aqueous extracts of P. niruri was found to be more effective in dissolution of CaOx with 63.33 % ± 1.44. P. niruri significantly (P < 0.05) inhibited the slope of nucleation and aggregation of CaOx crystallization, and reduced the crystal density. The results of the present study confirmed that P. niruri leaves can be used as remedial mediator for urolithiasis. However, further studies are required for isolation and identification of active constituents and their in-vivo confirmation.

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Investigation of potential anti-urolithiatic activity from different types of Musa pseudo-stem extracts in inhibition of calcium oxalate crystallization

BMC Complementary Medicine and Therapies ◽

10.1186/s12906-020-03113-0 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Mazni Abu Zarin ◽

Joo Shun Tan ◽

Paramasivam Murugan ◽

Rosma Ahmad

Keyword(s):

Calcium Oxalate ◽

Incubation Time ◽

Positive Control ◽

Negative Control ◽

Aggregation Assay ◽

Banana Tree ◽

Methanolic Extracts ◽

Different Types ◽

Aggregation Activity

Abstract Background The banana or scientifically referred to as Musa sp., is one of the most popular fruits all over the world. Almost all parts of a banana tree, including the fruits, stem juice, and flowers are commonly used as traditional medicine for treating diarrhoea (unripe), menorrhagia, diabetes, dysentery, and antiulcerogenic, hypoglycemic, antilithic, hypolipidemic conditions, plus antioxidant actions, inflammation, pains and even snakebites. The study carried out was to evaluate in vitro anti-urolithiatic activity from different types of Musa pseudo-stems. Methods Observing anti-urolithiathic activity via in vitro nucleation and aggregation assay using a spectrophotometer followed by microscopic observation. A total of 12 methanolic extracts were tested to determine the potential extracts in anti-urolithiasis activities. Cystone was used as a positive control. Results The results manifested an inhibition of nucleation activity (0.11 ± 2.32% to 55.39 ± 1.01%) and an aggregation activity (4.34 ± 0.68% to 58.78 ± 1.81%) at 360 min of incubation time. The highest inhibition percentage in nucleation assay was obtained by the Musa acuminate x balbiciana Colla cv “Awak Legor” methanolic pseudo-stem extract (2D) which was 55.39 ± 1.01%at 60 min of incubation time compared to the cystone at 30.87 ± 0.74%. On the other hand,the Musa acuminate x balbiciana Colla cv “Awak Legor” methanolic bagasse extract (3D) had the highest inhibition percentage in the aggregation assay incubated at 360 min which was obtained at 58.78 ± 1.8%; 5.53% higher than the cystone (53.25%).The microscopic image showed a great reduction in the calcium oxalate (CaOx) crystals formation and the size of crystals in 2D and 3D extracts, respectively, as compared to negative control. Conclusions The results obtained from this study suggest that the extracts are potential sources of alternative medicine for kidney stones disease.

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The Protective Roles of Estrogen Receptor β in Renal Calcium Oxalate Crystal Formation via Reducing the Liver Oxalate Biosynthesis and Renal Oxidative Stress-Mediated Cell Injury

Oxidative Medicine and Cellular Longevity ◽

10.1155/2019/5305014 ◽

2019 ◽

Vol 2019 ◽

pp. 1-17 ◽

Cited By ~ 3

Author(s):

Wei Zhu ◽

Zhijian Zhao ◽

Fu-Ju Chou ◽

Li Zuo ◽

Tongzu Liu ◽

...

Keyword(s):

Oxidative Stress ◽

Estrogen Receptor ◽

Nadph Oxidase ◽

Calcium Oxalate ◽

Cell Lines ◽

Ros Production ◽

Crystal Deposition ◽

Caox Crystal

Females develop kidney stones less frequently than males do. However, it is unclear if this gender difference is related to altered estrogen/estrogen receptor (ER) signaling. Here, we found that ER beta (ERβ) signals could suppress hepatic oxalate biosynthesis via transcriptional upregulation of the glyoxylate aminotransferase (AGT1) expression. Results from multiple in vitro renal cell lines also found that ERβ could function via suppressing the oxalate-induced injury through increasing the reactive oxygen species (ROS) production that led to a decrease of the renal calcium oxalate (CaOx) crystal deposition. Mechanism study results showed that ERβ suppressed oxalate-induced oxidative stress via transcriptional suppression of the NADPH oxidase subunit 2 (NOX2) through direct binding to the estrogen response elements (EREs) on the NOX2 5′ promoter. We further applied two in vivo mouse models with glyoxylate-induced renal CaOx crystal deposition and one rat model with 5% hydroxyl-L-proline-induced renal CaOx crystal deposition. Our data demonstrated that mice lacking ERβ (ERβKO) as well as mice or rats treated with ERβ antagonist PHTPP had increased renal CaOx crystal deposition with increased urinary oxalate excretion and renal ROS production. Importantly, targeting ERβ-regulated NOX2 with the NADPH oxidase inhibitor, apocynin, can suppress the renal CaOx crystal deposition in the in vivo mouse model. Together, results from multiple in vitro cell lines and in vivo mouse/rat models all demonstrate that ERβ may protect against renal CaOx crystal deposition via inhibiting the hepatic oxalate biosynthesis and oxidative stress-induced renal injury.

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Combination of garlic - shatterstone herb powder to control Streptococcus agalactiae infection in tilapia

Jurnal Akuakultur Indonesia ◽

10.19027/jai.14.79-89 ◽

2015 ◽

Vol 14 (1) ◽

pp. 79

Author(s):

Ririn Nurul Fauziah ◽

Dinamella Wahjuningrum ◽

, Sukenda ◽

, Ranta

Keyword(s):

Growth Rate ◽

Streptococcus Agalactiae ◽

Allium Sativum ◽

Positive Control ◽

Quality Parameters ◽

Negative Control ◽

Fish Pathogen ◽

Phyllanthus Niruri

ABSTRACT This study was aimed at determining potential of combination powder of garlic Allium sativum-shatterstone herb Phyllanthus niruri supplemented in feed against S. agalactiae infection in tilapia. Four concentrations of combination powder of A. sativum-P. Niruri; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in in vitro test. In vivo test consisted of three treatments with three replications, namely positive control (K+), negative control (K-) and the treatment of A. sativum-P. niruri suplemented in feed (BM). The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL S. agalactiae at concentration of 105 cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM) showed better growth rate, feed response, and survival (83.3%) than positive control (36.7%) at P<0.05. In addition, A. sativum-P. niruri suplemented in feed was also able to enhance the immune response by increasing phagocytic activity. Keywords: Streptococcus agalactiae, phytopharmacy, Allium sativum-Phyllanthus niruri, tilapia ABSTRAK Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih Allium sativum-meniran Phyllanthus niruri dalam pakan terhadap pencegahan infeksi bakteri S. agalactiae pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji in vitro. Uji in vivo terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif, kontrol negatif, dan perlakuan pakan yang mengandung bawang putih-meniran (BM). Uji ini dilakukan pada ikan nila berbobot 10,33±1,63 g yang dipelihara di akuarium dengan kepadatan 10 ekor/akuarium. Ikan diberi pakan perlakuan selama 14 hari kemudian diinjeksi secara intraperitoneal dengan bakteri S. agalactiae sebanyak 0,1 mL dengan kepadatan 105 cfu/mL pada perlakuan kontrol positif dan perlakuan BM. Parameter kelangsungan hidup, laju pertumbuhan, respons pakan, parameter hematologi, dan kualitas air diamati selama sepuluh hari. Hasil dari penelitian ini menunjukkan bahwa pemberian BM dalam pakan memberikan laju pertumbuhan, respons pakan, dan sintasan (83,3%) yang lebih baik daripada kontrol positif (36,7%) pada P<0,05. Pakan yang mengandung campuran bawang putih-meniran ini juga mampu meningkatkan respons imun dengan adanya peningkatan aktivitas fagositosis. Kata kunci: Streptococcus agalactiae, fitofarmaka, Allium sativum-Phyllanthus niruri, ikan nila

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Small intestine resection increases oxalate and citrate transporter expression and calcium oxalate crystal formation in rat hyperoxaluric kidneys

Clinical Science ◽

10.1042/cs20200973 ◽

2020 ◽

Vol 134 (19) ◽

pp. 2565-2580

Author(s):

Yi-Shiou Tseng ◽

Wen-Bin Wu ◽

Yun Chen ◽

Feili Lo Yang ◽

Ming-Chieh Ma

Keyword(s):

Small Intestine ◽

Calcium Oxalate ◽

Cell Injury ◽

Crystal Formation ◽

Crystal Deposition ◽

Tubular Cells ◽

Citrate Transporter ◽

Caox Crystal

Abstract Short bowel (SB) increases the risk of kidney stones. However, the underlying mechanism is unclear. Here, we examined how SB affected renal oxalate and citrate handlings for in vivo hyperoxaluric rats and in vitro tubular cells. SB was induced by small intestine resection in male Wistar rats. Sham-operated controls had no resection. After 7 days of recovery, the rats were divided into control, SB (both fed with distilled water), ethylene glycol (EG), and SB+EG (both fed with 0.75% EG for hyperoxaluric induction) groups for 28 days. We collected the plasma, 24 h of urine, kidney, and intestine tissues for analysis. Hypocitraturia was found and persisted up to 28 days for the SB group. Hypocalcemia and high plasma parathyroid hormone (PTH) levels were found in the 28-day SB rats. SB aggravated EG-mediated oxalate nephropathy by fostering hyperoxaluria and hypocitraturia, and increasing the degree of supersaturation and calcium oxalate (CaOx) crystal deposition. These effects were associated with renal up-regulations of the oxalate transporter solute carrier family 26 (Slc26)a6 and citrate transporter sodium-dependent dicarboxylate cotransporter-1 (NaDC-1) but not Slc26a2. The effects of PTH on the SB kidneys were then examined in NRK-52E tubular cells. Recombinant PTH attenuated oxalate-mediated cell injury and up-regulated NaDC-1 via protein kinase A (PKA) activation. PTH, however, showed no additive effects on oxalate-induced Slc26a6 and NaDC-1 up-regulation. Together, these results demonstrated that renal NaDC-1 upregulation-induced hypocitraturia weakened the defense against Slc26a6-mediated hyperoxaluria in SB kidneys for excess CaOx crystal formation. Increased tubular NaDC-1 expression caused by SB relied on PTH.

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ANTIUROLITHIATIC ACTIVITY OF NATURAL CONSTITUENTS ISOLATED FROM AERVA LANATA FLOWERS

International Journal of Pharmaceutical Sciences and Drug Research ◽

10.25004/ijpsdr.2020.120610 ◽

2020 ◽

pp. 668-673

Author(s):

Subhashini Naikal James Prameela ◽

Shilpika Nagula

Keyword(s):

Crystal Growth ◽

Calcium Oxalate ◽

High Performance ◽

Crystal Nucleation ◽

Aggregation Assay ◽

Alcohol Extract ◽

Active Constituents ◽

Caox Crystal ◽

Aerva Lanata

A well-known traditional herb Aerva lanata, broadly used in India for treatment of different ailments such as urolithiasis. Pashanabheda is used as antiurolithiatic in Ayurveda. In the present study, flowers of A. lanata were selected for isolation of active constituents and screening for in vitro antiurolithiatic potentials, as literature supports that flowers have the highest quantity of natural components when compared with the other parts. Hydroalcoholic (80%-water, 20%-alcohol) extract of A. lanata flowers was prepared and fractionation with different organic solvents.. The two fractions (ethyl acetate and n-butanol) were subjected to isolation of active constituents using column chromatography technique, followed by purification of the isolated constituents by preparative high performance thin layer chromatography (HPTLC)and then the individual components were characterized by IR spectrophotometery. Finally, in vitro antiurolithiatic activity was screened by nucleation and aggregation assay. In the aggregation assay, gradual decrease in the calcium oxalate (CaOx) crystal nucleation as well as growth was observed by light microscopy. The findings of the nucleation assay indicate that phytoconstituents inhibited the crystallization of CaOx in solution. The size and the number of calcium oxalate crystals decreased with increasing concentration of the phytoconstituents. The increasing concentrations of Quercetin and betulin (100, 200, 300, 400 and 500 μg/mL) inhibited the CaOx crystal growth. The isolated quercetin and betulin from A. lanata have shown antiurolithiatic effect by significantly reducing the CaOx crystal growth.

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Temporal Changes in mRNA Expression for Bikunin in the Kidneys of Rats during Calcium Oxalate Nephrolithiasis

Journal of the American Society of Nephrology ◽

10.1681/asn.v105986 ◽

1999 ◽

Vol 10 (5) ◽

pp. 986-996

Author(s):

SHIZUKA IIDA ◽

AMMON B. PECK ◽

JOANNE JOHNSON-TARDIEU ◽

MANABU MORIYAMA ◽

PATRICIA A. GLENTON ◽

...

Keyword(s):

Ethylene Glycol ◽

Calcium Oxalate ◽

Mrna Expression ◽

Kidney Tissue ◽

Positive Control ◽

Renal Tubular ◽

Competitive Template ◽

Rat Kidneys

Abstract. Inter-α-inhibitor and other bikunin-containing proteins are synthesized in relatively large quantities by the liver. These proteins function as Kunitz-type serine protease inhibitors and appear capable of inhibiting calcium oxalate (CaOx) crystallization in vitro. Preliminary studies have shown that renal tubular epithelial cells synthesize bikunin in response to CaOx challenge. To examine this response in vivo, a sensitive reverse transcription-quantitative competitive template-PCR was developed to detect and quantify poly(A)+ -tailed bikunin mRNA expression in kidney tissue from normal rats and rats developing CaOx nephrolithiasis after challenge with ethylene glycol. Bikunin mRNA expression in rat liver tissue was assessed as a positive control. The expression of bikunin mRNA in liver did not differ significantly between normal control rats and experimental rats with induced hyperoxaluria and renal CaOx crystallization. In contrast, there were significant temporal increases in the levels of bikunin mRNA expression in rat kidneys during CaOx nephrolithiasis after challenge with ethylene glycol. Urinary excretion of bikunin-containing proteins seemed to increase concomitantly. These findings indicate an association between the induction of hyperoxaluria/CaOx nephrolithiasis and the expression of the bikunin gene in rat kidneys.

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In Vitro and In Vivo Neutralizing Activity of Uvaria chamae Leaves Fractions on the Venom of Naja nigricollis in Albino Rat and Bovine Blood

Recent Patents on Biotechnology ◽

10.2174/1872208314666200903152129 ◽

2020 ◽

Vol 14 (4) ◽

pp. 295-311

Author(s):

Ada Gabriel ◽

Mamman Mohammed ◽

Mohammed G. Magaji ◽

Yusuf P. Ofemile ◽

Ameh P. Matthew ◽

...

Keyword(s):

Ethyl Acetate ◽

Neglected Tropical Diseases ◽

Lethal Dose ◽

Positive Control ◽

Haemolytic Activity ◽

Cytotoxic Activities ◽

Albino Rat ◽

Aqueous Residue

Background: Snakebite envenomation is a global priority ranked top among other neglected tropical diseases. There is a folkloric claim that Uvaria chamae is beneficial for the management of snakebite and wounds in African ethnobotanical surveys. Besides, there are many registered patents asserting the health benefits of U. chamae. Objective: This study aimed to investigate U. chamae’s potentials and identify candidates for the development of tools for the treatment and management of N. nigricollis envenomation. Methods: Freshly collected U. chamae leaves were air-dried, powdered, and extracted in methanol. The median lethal dose of the extract was determined and further fractionated with n-hexane, n-butanol and ethyl acetate. Each fraction was tested for neutralizing effect against venom-induced haemolytic, fibrinolytic, hemorrhagic, and cytotoxic activities. Results: U. chamae fractions significantly (p<0.05) neutralized the haemolytic activity of N. nigricollis venom in n-butanol; 31.40%, n-hexane; 33%, aqueous residue; 39.60% and ethyl acetate; 40.70% at the concentration of 100mg/ml of each fraction against 10mg/ml of the snake venom when compared to the positive control. The fibrinolytic activity of N. nigricollis venom was significantly (p<0.05) neutralized in n-hexane at 73.88%, n-butanol; 72.22% and aqueous residue; 72.22% by the fractions of U. chamae. In addition, haemorrhagic activity of N. nigricollis venom was significantly (p<0.05) neutralized by U. chamae fractions at the concentrations of 100mg/ml, 200mg/ml and 400mg/ml except for n-butanol and aqueous residues at 400 mg/ml. Conclusion: U. chamae leaves fractions possess a high level of protection against N. nigricollis venoms-induced lethality and thus validate the pharmacological rationale for its usage in the management of N. nigricollis envenomation.

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Therapeutic effects of sesamolin on leukemia induced by WEHI-3B in model mice

Applied Biological Chemistry ◽

10.1186/s13765-021-00616-3 ◽

2021 ◽

Vol 64 (1) ◽

Author(s):

Senthil Nagarajan ◽

Jae Kwon Lee

Keyword(s):

Nk Cell ◽

Neoplastic Cell ◽

Positive Control ◽

Cytolytic Activity ◽

Leukemic Cells ◽

Therapeutic Effects ◽

Control Drug ◽

Lysis Activity

AbstractSesamolin is one of the lignans derived from sesame oil. It has demonstrated significant antioxidant, anti-aging, and anti-mutagenic properties. It also reportedly augments natural killer (NK) cell lysis activity. We previously reported that sesamolin also exerts anticancer effects in vitro and induces enhanced NK cell cytolytic activity against tumor cells. Herein, we aimed to determine the mechanism by which sesamolin prevents and retards tumorigenesis in BALB/c mouse models of leukemia induced by murine (BALB/c) myelomonocytic leukemia WEHI-3B cells. Banded neutrophils, myeloblasts, and monocytic leukemic cells were more abundant in the leukemia model than in normal mice. Sesamolin decreased the number of leukemic cells by almost 60% in the leukemia model mice in vivo; additionally, sesamolin and the positive control drug, vinblastine, similarly hindered neoplastic cell proliferation. Spleen samples were ~ 4.5-fold heavier in leukemic mice than those obtained from normal mice, whereas spleen samples obtained from leukemic mice treated with sesamolin had a similar weight to those of normal mice. Moreover, sesamolin induced a twofold increase in the cytotoxic activity of leukemic mouse NK cells against WEHI-3B cells. These results indicated that sesamolin exerts anti-leukemic effects in vivo.

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Mutagenicity evaluation of Anastatica hierochuntica L. aqueous extract in vitro and in vivo

Experimental Biology and Medicine ◽

10.1177/1535370217748574 ◽

2017 ◽

Vol 243 (4) ◽

pp. 375-385 ◽

Cited By ~ 1

Author(s):

Siti Rosmani Md Zin ◽

Zahurin Mohamed ◽

Mohammed A Alshawsh ◽

Won F Wong ◽

Normadiah M Kassim

Keyword(s):

Aqueous Extract ◽

Positive Control ◽

In Vitro Assay ◽

In Vivo Study ◽

Sufficient Evidence ◽

Aqueous Extracts ◽

Mutagenic Potential ◽

Vitro Assay

Anastatica hierochuntica L. ( A. hierochuntica), a folk medicinal plant, was evaluated for mutagenic potential via in vitro and in vivo assays. The in vitro assay was conducted according to modified Ames test, while the in vivo study was performed according to Organisation for Economic Co-operation and Development guideline for mammalian erythrocyte micronucleus assay. Four groups ( n= 5 males and 5 females per group) Sprague Dawley rats were randomly chosen as the negative control, positive control (received a single intramuscular injection of cyclophosphamide 50 mg/kg), 1000 and, 2000 mg/kg A. hierochuntica aqueous extracts. All groups except the positive control were treated orally for three days. Findings of the in vitro assay showed mutagenic potential of AHAE at 0.04 and 0.2 mg/ml. However, no mutagenic effect was demonstrated in the in vivo study up to 2000 mg/kg. No significant reduction in the polychromatic and normochromatic erythrocytes ratio was noted in any of the groups. Meanwhile, high micronucleated polychromatic erythrocytes frequency was seen in cyclophosphamide-treated group only. These findings could perhaps be due to insufficient dosage of A. hierochuntica aqueous extracts to cause genetic damage on the bone marrow target cells. Further acute and chronic in vivo toxicity studies may be required to draw pertinent conclusion on the safety aspect of A. hierochuntica aqueous extracts consumption. Impact statement In this paper, we report on the mutagenicity evaluation of Anastatica hierochuntica aqueous extract. This is a significant research in view of the popularity of this herb consumption by the people across the globe despite of limited scientific evidence on its toxicity potential. This study is intended to encourage more extensive related research in order to provide sufficient evidence and guidance for determining its safe dosage.

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