The present research investigation intends at investigating the effect of on Sialic acid components in male albino rats of an aqueous extract of seeded papaya fruit pulp and seed extract specified as a combined dose and seedless variety papaya fruit pulp extract. The primary objective of the current research investigation was to evaluate the effects that occurred on male albino Wistar rats of semi-ripe fruit pulp extract from Carica papaya (fruits with and without seeds). The development of spermatozoa's ability to fertilise, the coating of spermatozoa with specific antigens, and changes in their membrane surface are all related to Sialic acid. Less spermatozoa cells in the testicular compartments and fewer of these cells are indicated by the decrease in Sialic acid contents in the testis and epididymis of both experimental groups

Manivelu D.

Punitha N.

Selvi  P.V.

Subramanian N.

Sudha M.

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Journal of Stress Physiology & Biochemistry, Vol. 20, No. 2, 2024, pp. 14-19     ISSN 1997-0838Original Text Copyright © 2024 by Punitha, Subramanian, Manivelu, Sudha and SelviORIGINAL ARTICLEStudies on the Impact of Semi-ripe along with SeedlessPapaya (Carica papaya) Fruits Pulp and Seeds Extract onthe Level of Sialic acid in male Albino Rat ReproductivetissuePunitha N.1, Subramanian N.1, Manivelu D.2, Sudha M.3and Selvi  P.V.31 Department of Zoology, Arignar Anna Govt. Arts College, Cheyyar, Thiruvannamalai District, Tamilnadu, India.2 Department of Zoology, Govt. Arts College for Men, Krishnagiri, Tamilnadu, India.3 Department of Zoology, Bharathi Women’s College, Chennai, Tamilnadu, India.*E-Mail:  prof.dmani@gmail.comReceived January 21, 2024The  present  research  investigation  intends  at  investigating  the  effect  of  on  Sialic  acidcomponents in male albino rats of an aqueous extract of seeded papaya fruit pulp and seedextract  specified  as  a  combined  dose  and  seedless  variety  papaya  fruit  pulp  extract.  Theprimary objective of the current research investigation was to evaluate the effects that occurredon male albino Wistar rats of semi-ripe fruit pulp extract from  Carica papaya (fruits with andwithout  seeds).  The  development  of  spermatozoa's  ability  to  fertilise,  the  coating  ofspermatozoa with specific antigens, and changes in their membrane surface are all related toSialic acid. Less spermatozoa cells in the testicular compartments and fewer of these cells areindicated  by  the  decrease  in  Sialic  acid  contents  in  the  testis  and  epididymis  of  bothexperimental groups.Key words:  Extracts, Carica papaya, Albino Rat, Testis, Epididymis, Sialic acidJOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 20  No. 2  2024Studies on the Impact of Semi-ripe ...India and other nations have a long history of usingmedicinal plant products for traditional purposes (Govil,1998). Papaya seed as well as peel–pulp has a potentialfor  pharmacological  raw  materials.  The  data  wouldprovide  fundamental  data  for  the  use  of  papayas  fornatural  antioxidants  also  (Yoon  et  al.,  2022).  Crudeextracts from seeds of Carica papaya have been shownto  induce  variable  responses  depending  on  the  doseduration  and  route  of  administration  in  laboratoryanimals (Udoh and Kehinde, 1999; Kamal  et al.,  2003;Verma and Chinoy, 2002). Papaya seeds have a longhistory  of  traditional  use  in  various  cultures  for  theirmedicinal properties (Sharma et al., 2022).While there is a lengthy history of  phytotherapy,  aproper  scientific  explanation  is  relatively  recent.Numerous  medicinal  herbs  both  domestically  andabroad  are  linked  to  antifertility  effects  (Duke,  1985).Papaya seed n-hexane fraction has potential as a malecontraceptive by reducing spermatogenesis activity anddecreasing sperm quality in white rats (Sri Nita  et al.,2019).  Epididymis plays a very important role in malereproduction.  The  epididymis  brings  the  sperms  tomaturity since the sperms that emerge from the testesare  immature  and  are  incapable  of  fertilization(Rajalakshmi, 1992).  Since the epididymal epithelium isin charge of producing Sialic acid and proteins that arepoured straight  into  the lumen,  it  is  essential  that  thespermatozoa  in  the  epididymis  be  exposed  to  themicroenvironment  of  the  organ  in  order  for  them  toacquire  the ability  to  fertilise and is  an active  process(Chinoy et al., 1995; Johnson et al., 2000). Sialic  acid,  which  covers  some  antigenic  markersand  makes  sperm  negatively  charged  to  preventaggregation,  is  directly  linked  to  sperm  maturation,capacitation,  and  sperm-egg  identification.  Theseprocesses boost the survival rate of sperm in the femalereproductive canal. It is commonly known that the seedsof the Carica papaya plant contain substances that mayregulate  fertility.  Testicular  secretory  components  likeglycogen,  protein,  cholesterol  and  Sialic  acid  can  beused to determine the functional capacity of the testes(Kamatchouing et al., 2002, Watcho et al., 2004) In rats,rabbits,  and  langur  monkeys,  a  preclinical,  briefassessment employing a range of extraction techniqueson the seeds of C. papaya demonstrated spermatogenicarrest,  resulting  in  azoospermia,  or  completesuppression  of  sperm  motility  (Lohiya  et  al.,  1994,2006).MATERIALS AND METHODSCarica papaya  fruits,  both seedless and semi-ripe,were purchased commercially from Chennai's wholesalefruit  market.  The  fruit's  outer  skin  and  interior  seedswere removed and they were then cleaned in double-distilled water. Fruit pulp was cut into slices. Separately,the  sliced  fruit  pulp  and  seeds  were  air  dried  in  theshade. Using an electronic blender, the dried chunks offruit pulp and seeds from the seeded variety and the fruitpulp from the seedless variety were separately groundinto  a  coarse  texture.  After  being  macerated  in  coldwater, the pulped fruit was filtered through a fine muslincloth.After  being  collected,  the  filtrate  was  dried.  Thedehydrated substance was stored in a plastic containerthat sealed tightly. For later usage, the dried fruit pulpand seed fractions were stored in a refrigerator at 40oC.Aqueous extracts of two distinct kinds were made. Therequired amount of distilled water was used to preparethe fruit pulp of the seedless variety after the fruit pulpand seed of the seeded variety were combined. Beforethe animals were given the extracts, a new sample ofthe necessary quantities was taken from the stock.The  King  Institute  of  Preventive  Medicine  andResearch, Guindy, Chennai, was the source of healthymale  and  female  albino  Wistar  strain  rats  (Rattusnorvegicus).  In  this  investigation,  rats  weighing 155 ±25g were employed. The animals were kept in regularenvironmental conditions with adequate ventilation anda temperature of 25±20º C in polypropylene cages witha metal grill top. They received water on demand andwere subjected to a 12-hour light–dark cycle.A regular pelleted meal that was balanced was fed tothe animals. Animals received humane treatment. Careand  oversight  were  given  during  the  course  of  theinvestigation. Acute oral toxicity studies were conductedin  accordance with  the  CPCSES recommendations  toevaluate the harmful effects, establish a tolerance limit,JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 20  No. 2  202415Punitha et al.and establish a safe dosage. Before the rats were giventhe extracts,  they were fasted for three hours.  Gastricintubation  was  used  to  give  the  extracts  as  a  singledosage. For every group, there were three sets of sixrats. For oral delivery equal parts of fruit pulp power andseeded variety powered seed were removed, blended,and  well  mixed  in  distilled  water.  The extract  mixturewas  administered  at  doses  of  1000mg,  2000mg,3000mg, 4000mg, and 5000mg / kg bw. The LD50 wascalculated  after  96  hours  of  observation  of  mortality(Weis, 1952).Every  day  at  9.00  am,  the  test  dose  wasadministered. After dosing, animals were first inspectedat least once in the first thirty minutes and then again onoccasion in the first twenty-four hours. The fatality ratesfor dosages of 5000mg, 4000mg, 3000mg, and 2000mgwere  100%,  67%,  50%,  and  33%,  respectively.  At  adosage  of  1000mg,  no  mortality  was  noted.  A  50%death  rate  was  noted  at  3000mg/kg  bw.  The  mixedpreparation was given orally to the rats in experimentalgroup I for duration of 60 days, after which a third of thesafe dose for the animals was ascertained.Testing  for  toxicity  and  tolerance  limits  was  alsodone  on  rats  given  papaya  fruit  pulp  of  the  seedlesskind. A dosage of 1000mg, 2000mg, 3000mg, 4000mg,and  5000mg/kg  bw  was  applied  to  the  pulp  extract.Mortality  rates  for  5000mg,  4000mg,  3000mg,  and2000mg  dosages  were  83%,  67%,  50%,  and  17%,respectively. At a dosage of 1000mg, no mortality wasnoted.  There  was  a  50%  death  rate  reported  in3000mg/kg bw. Rats in the experimental group  II weregiven the aqueous preparation orally for duration of 60days, with a third of the safe dose being fixed from this.Before  mending,  the  animals  were  watched  forindicators  of  toxicity  and  death  for  48  hours  aftertreatment.After determining the tolerance limit for both types ofextracts, the experiment's final doses were decided. Forthe animals in experimental group I, 500mg of dried fruitpulp powder and 500mg of powdered seed from semi-ripe  papaya  fruit  seeds  (a  total  of  1000mg)  weredissolved  in  water  (1.0gm  dissolved  in  1.0ml  distilledwater).  For  the  animals  in  experimental  group  II,1000mg of fruit pulp powder from seedless papayas wasdissolved  in  water  (1.0gm  dissolved  in  1.0ml  distilledwater).  Throughout  the  course  of  the  treatment,  theanimals in each group were given the same dose.The synthesized extracts were given orally throughstomach intonation through the use of an orogastric tubethat  included  a  16-G  polyethylene  catheter  that  wasequipped  with  a  10-ml  hypodermic  syringe.  For  sixtydays in a row, extracts were administered every morningfollowing a 24-hour gap.Experimental designWhenever  the  animals  were  weighed,  they  werecategorized  into  three  groups  on  the  basis  of  theirweight. Each group featured six animals that were keptin a variety of cages.Group  I:  Control: Male  animals  which  gotnourishment and hydrated appropriately.Group  II:  Experimental  group-I:  For  an  entireperiod of 60 days, oral administration of seeded semi-ripe papaya fruit pulp and seed extract (1000mg/kg bodyweight/day)  has  been  given  to  experimental  maleanimals, in conjunction with their usual diet of food andwater.Group  III:  Experimental  group-II:  For  an  entireperiod of 60 days, oral administration of seedless, semi-ripe  papaya  fruit  pulp  extract  (1000mg/kg  bodyweight/day)  has  been  given  to  experimental  maleanimals, in conjunction with their usual diet of food andwater.Before  the  trial  began,  the  animals  were  givenregular  meal  and water for  15 days to  help them getused to the lab environment. The starting body weightswere noted both before and after the intervention. Everyday at  9.00 a.m.,  the semi-ripe papaya fruit  pulp  andseed extract with seeds and the seedless variety's semi-ripe papaya fruit  pulp extract  were administered orallyvia stomach intubation. 24 hours after the final dose wasadministered,  on  the  61st day,  the  animals  weresacrificed by cervical dislocation.The Warren (1959) method was used to estimate theamount of Sialic acid. The data was presented as Mean± SD and analysed using one way analysis of variance(ANOVA), multiple range tests, and Duncan's test. UsingSPSS version 6.0, the statistical significance was testedJOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 20  No. 2  202416Studies on the Impact of Semi-ripe ...at 1% and 5% levels.RESULTS AND DISCUSSIONThe concentrations  of  Sialic  acid  in  the  testis  andepididymal  tissues  of  male  rats  provided  papaya  fruitpulp and seed extracts are shown in the given Table andFig. The first experimental group's testis and epididymisSialic acid contents exhibited a substantial decline (p <0.001). When compared to control animals, the secondexperimental  group's  testis  revealed  a  drop,  but  itsepididymis did not change markedly.Table 1:  Effect of Papaya semi-ripe fruit pulp and seed extracts on Sialic acid content in male albino rats. TissuesControl -Normalfeed and waterExperimental Group –I Seeded papaya -Fruit pulp and seedextract combinedExperimental Group -IISeedless papaya-Fruit pulp extractF- value P- valueTestis 7.04 c ± 0.25 5.80 a ± 0.26 6.35 b ± 0.22 969.159 < 0.001**Epididymis 8.69 b ± 0.18 6.72 a ± 0.01 8.21 b ± 0.17 25.681 < 0.001**Values are expressed as mg/gm wet tissue; Values are Mean ± SD (n=6) observations;              ** denotes significanceat 1% level;  Different alphabets between groups denotes significance at  5% level using Duncan multiple range test(DMRT); Means carrying at least one common superscript between groups do not differ significantly.Figure 1. Effect of Carica papaya semi-ripe fruit pulp and seed extract on sialic acid content in male albino rats.Sialic  acid  has  roles  in  the  development  ofspermatozoa's  propensity  to  fertilise,  the  alteration  oftheir membrane surface when they mature and and thecoating of spermatozoa with specific antigens (Chauhanand  Agarwal,  2009).  Less  spermatozoa  cells  in  thetesticular  compartments  and  fewer  male  reproductivecells in the epididymal lumen have been shown by thedecrease  in  Sialic  acid  contents  in  the  testis  andepididymis  of  both  experimental  groups.  In  order  toenhance  the  mobility  of  sperm  and  diminish  frictionJOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 20  No. 2  202417Punitha et al.between  spermatozoa,  Sialic  acid  performs  as  alubricant (Riar et al., 1973).The result are in agreement with those of Riar et al,(1973) they  also  founded  similar  result.  The  non-maturing of  the sperm is triggered by perturbations inthe  biochemical  mileu  of  the  epididymis  (Verma  andChinoy, 2002). There have been no obvious alterationsrevealed in the epididymis about the animals in Secondexperimental group.CONCLUSIONThe present study was investigated at the effect ofon Sialic  acid  components  in  male  albino  rats  of  anaqueous  extract  of  seeded  papaya  fruit  pulp  andseedless papaya fruit pulp. The main objective of the current research was toevaluate the effects that occurred on male Albino Wistarrats of semi-ripe fruit pulp extract from Carica papaya. The  development  of  spermatozoa's  ability  tofertilise,  the  coating  of  spermatozoa  with  specificantigens, and changes in their membrane surface areall related to Sialic acid. Less  spermatozoa  cells  in  the  testicularcompartments and fewer of these cells are indicated bythe decrease in Sialic  acid contents in the testis andepididymis of both experimental groups.CONFLICTS OF INTERESTThe  authors  declare  that  they  have  no  potentialconflicts of interest. REFERENCESChauhan  A  and  Agarwal.  M  (2009)  Pharmacologicalevaluation for the antifertility effect of the ethanolicseed extract  of  Nelumbo nucifera  (Sacred  lotus).Pharmacology Online, 2 :636-643.Chinoy  N.J.D.,  Souza.  J.M  and  Padman.  P.  (1995)Contraceptive  efficacy  of  Carica  papaya  seedextracts  in  male  albino  mice  (Mus  musculus).Phytother Res., 9:30-36.Duke J.A. (1985) Medicinal plants. Science. 229: 1036-1038.Govil  J.N.  (1998)  Current  concepts  of  multidisciplineapproach to  the  Medicinal  plants  (part  I),.  Todayand  Tomorrow’s  Printers  and  Publishers,  NewDelhi, Vol. XII ,pp.1-28.Johnson  L.,  Varner.  D.D.,  Roberts.  M.E.,  Smith.T.L.,Keillor. 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Studies on the Impact of Semi-ripe along with Seedless Papaya (Carica papaya) Fruits Pulp and Seeds Extract on the Level of Sialic acid in male Albino Rat Reproductive tissue

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Studies on the Impact of Semi-ripe along with Seedless Papaya (Carica papaya) Fruits Pulp and Seeds Extract on the Level of Sialic acid in male Albino Rat Reproductive tissue

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