International Journal of Biological Chemistry 1 (4): 221-224 ISSN 1819-155, © 2007 Academie Jounal Ine. 2007 Biopotency of erva lanata on Membrane Bound ATPases and Marker Enzymes in Urolithie Rats P, Soundararajan, R. Mahesh, T. Ramesh and V. Hazeena begum Department of Sidahu Medicine, Faculty of Science, Tamil University, ‘Thanjavur, Tamilnadu-613 010, India Abstract: 4 lanaraequeces suspension was tested for its biopoteney on membrane bound enzymes and marker enzymes in rolithisssin male albino rats, Caleium oxalate wolithiasis as induced by 0.75% alilene glyco! in drinking water for 28 days. There was significant ‘decrease in membrane bound enzyines such as Na'K'ATPase, Ca!"ATPase, Mg ATPase snd marker enzymes Aspartate Transaminase (AST), Alanine Transaminase (ALT), Alkaline Phosphatase (ALP) and Gamma-Glutamyt Transferase (GGT) in kidney and liver while Lactate Dehydrogenase (LDH) was increased In serum electalytes, sodium was reduced snd K, Ca and Mg levels wete elevated, The AST. ALT, ALP, LDH and GGT were increasedin serum and usin: of wolithie rats, Therapeue reahment with A. nara aqusous suspension (2 g kg Dwt dose-* day-* oral~) ls significantly ameliorated to near normalcy in the curative group. The results showed that 4. fanate might represent a alternative form of treatment andor prevention of wolithiasis, Key words Urolithiasis, membrane bound ATPases, marker enzymes, lanata INTRODUCTION Eiiylne glycol was reported to induce calcium oxalate type of urlithiass by the formation of ‘oxalate, becaute ofthe intermediates ofplycclate and glyoxylate in thera liver (Halabe eta, 2003, Poors ef a, 1997), Oxalate causes peroxidation of membrane lipids andl oxidation of protsins, initiates ‘the loss of membrane integrity ane activities of membrane bound enzymes (Selva, 2002). Membrane ATPases may play an important role in the maintenance of ionic gradients between the intracellular! extracellular compartments of th cell. Membrane Na°k’ATPase plays an important role in aetive ‘tansport of Na" and K* ions across the plasma membrane simi, Ca"*ATPase is clearly linked with Ca* pump and transport of Ca" ions, while Mg” activated ATPase is distributed in all renal cell compartments. The Mg ion forms Mg*ATP complex, which is the subsite for the enzyme. Mp* ATPase is to conisol the intracellular Mg concentration, changes which can modulate the activity of Me” dependent enzymes and regulate rates of protein synthesis and os growth (Stekhoren ‘and Renting, 1981), Lipid peroxidation leads to cell damage leading to the release of marker enzymes namely Aspartate Transaminase (AST), Alanine Transaminase (ALT), Alkaline Phosplutase (ALP), [Lactate Dehydrogenase (LDH and Gammu-Glutamy Transferase (GGT) into blood vireulatin and ‘wine, Most of the urinary enzymes crginating in the kidneys ae localized to specific regions and cellular componnis of the nephron, thereby studies pertaining to these enzymes will show the pathological stats of the kidney (Srinivasan er af, 2004), Current-day medical management of ‘urolithissis mainly involves the surpical removal of stones by the techniques such as ESWL (Extracorporeal shockwave lithotripsy), PCNL (Pereutanzous neplwolithotomy), but they donct assure the prevention of recurrence of the stone. They cause side effects such ‘Carresponaing Author: De V-Hateoa Ream Deparment of Sita Medios, Fac of Sece, Taal UWaR, ‘Vey, Thayjvur 61 10, Ta, oda 21Int Biol. Chem, 1 (8): 22 5, 20 as heamorthage, hypertension, tubular necrosis and subsequent fibrosis ofthe kidney, Hence, the search of effective herbal. drugs forthe treatment of urlithiass with no side effets is necessary _Aerva Janata is herbaceous weed, ound throughout the hotter parts ofthe India especially in the sates of Tamilnadu, Andhra Pradesh and Kamataka,t, anata is endowed with various chemical ‘components sch as flavonoids, alkaloids, steroid, polysaccharides, tamins, saponins, ete. (Chandra and Sastry, 1990; Afog er a, 1991; Zapesochaya er af, 1992), which possibly contribute to its diverse uses in folklore medicine. Also 4. fanata was reported to have ant-nflammatory, diuretic (Vetrichelvan er af, 2000), antimicrobial, eytotonicity (Chowdhury er af, 2002), antidiabetic (Vetichelvan and Jepadeesan, 2002) and nephreprotective(Shiswaikur ea, 2004) activities. In our previous studies, . anata reduced the oxalate synthesizing enzymes, oxalate andl urinary risk factors and lipid in etivene lye induced wolithiass (Soundararajan eta, 2006, 2007), The present study vas planned to evaluate the potential of 4. fanata aqueous suspension on membrane bound ATPases ‘nkichey and liver and marker eneymes in kidey, liver, sera and urine of etylene glycol induced urolithiasis in rats MATERIALS AND METHODS. Plant Material Fresh. fanata aerial parts were collested during the months of September to December 2003 ‘The aerial pasts were dried thocoughly under shade and powdered finely. The fine powder was suspended a concentration of $00 mg miL~ with distilled water as aqueous suspension and used for the study. Animals Male albino rats of Wistar strains weighing approximately 140-150 g were wed, All animal experiments ond muintenaneo wero carried out in our laboratory accerding. to the cthical auidelines suggested by the Insitutonal Animal Ethies Committee after Famuary 2004. Animals were housed in polypropylene cages under controlled conditions of 12 fh light 12 h dark cycle and temperature 272°C. All of the ats received standard pellet diet (Amrut rat feed, Pune) and water ad bina, ‘Stone Induction _Aninoals Were Fed with 0.75% ellyleneelvodl in dining water for 28 days ad lbita to induce the wolihiasis. The uroithie rats were then used for the stu Experimental Design In the experiment total of 24 rats (12 welt rts, 12 contd suis) were sed, The sats were divided in to-4 groups of 6 rts each, Group I served as contre. Group I served as dg control which received A Janata aquecus suspension alone (2 8 ke~! bawt. dose! day oral) foe 28 days, Group UL served as urolithc sats. Calcium oxalate urolithio rats were trated with 4, Janata aquzcus suspension (2g kg bowt dose! day! oral~) for 28 days (Group IV) “Atthe end of 28 days, the animals were housed in metabolio cages and 24 h urine was collected. After the urine collection, the animals were anaesthetized with Pentobarbitol sodium (35 mg kg~* bodyweight, ip). Blood was drawn fom the external jugular vein and serum was separated by -cntrifugaton at 3000 rpm. Kidneys and liver were excised and immersed in iee-cold physiological saline ancl blotted with filter paper. Known weight of tissues were homogerized in 0.1 M tris HCL ‘butler pH 7.4 containing 0.25 M suetose and used for biochemical estimations. m2Int Biol. Chem, 1 (8): 22 Biochemical Assays ‘The total homogenate was used for assaying membrane bound enzymes like Na'K "ATPase (Bonting, 1970), Ca ATPase (Hierten and Pan, 1983), Mg™ ATPase (Ohnishi era, 1982) activities. ‘The amount of inorganic phosphorus was determined by the method of Fiske and Subbarow (192: Serum sodium, potassium, calcium and magnesium were analysed by using he methods of Tinder (1951), Mamnna (1958), Gitelman (1967) and Neil and Neely (1956), respectively. The activities of AST (Mohun and Cook, 1957), ALT (Motun snd Cook, 1957), ALP (Kind and King, 1954), LDH (King, 1965) and GGT (Persijin and vander Sik, 1976) in kidney, liver, serum and 24h utine were assayed. ‘Statistical Analysis "The results were presented asthe meantSD. One-way analysis of variance (ANOVA) flowed by the Tukey’s test for multiple comparisons was used. Values of p<0.05 was considered to be significant. SPSS 7.5 version was used fr this analysis. RESULTS: ‘The aotivty of Na°K'ATPase, Ca ATPase, Mg ATPase were significantly diminished in ethylene glyco! induced urolitic rats (Group 111) when compared with contr rats (Group D. The activity ofthese enzymes was reverted to near normal in. Janata aqucous suspension administered sats (Group TV). No significant change was observed in 4. fanota aquecus suspension treated drug contol rats (Group ID) as compared fe contol group (Group 1) (Table 1), ‘The level of Na’ was markedly lowered while K’, Ca and Mg” levels were clevated in ethylene glycol induced urolihie rats (Group IID) when compared to contol rats (Group 1. Administration of 4. farata aqueous suspension brought the Na‘, K°,Ca* and Mg levels near 10 ‘ible set of nai onmemtrane bound ATPases in onl a experimental rape NFR'ATPase CATR MeATPRE perimental Drie dosge : = ‘gh * at es f gare phonons Wert mg pete) ianey 1 ase azainot Lasoo 0 2 Sstsn10 227009 Lsunor 1 anor L2sa08 nssoore 7 2 stom 2ieoor Lassa 1 2am s9un07 Lasioo8 0 Dato Hain Lasanoe m z Lennon Uneats osss900" w 2 22run0s 30 Ls2u008" ‘Vals ae exposed af nar Te als each rp ay Cope ih arp 1 Coie ith 3m 1.» p0.001 ‘able 2: sft of ana om seu sodium, pots, clea an magnesia in cota nd xperonees arog Drug desge Sadun Pets Caleam “Magnes ibe) rong dL“) s26soeea 140%0 7 9040.83 2srant9 : pedsame sang =O ManSPe STN sigsusate swat ___tomsnaite rsa ‘Vals are eres a meaSD fr 6 inals enh grow.” as coped wah group T= corgael wih arvep TL *:p-0.001 223