The results of the present investigation suggested the antioxidant effects of the JDEE against the oxidative stress induced with CCl4 in testes of male rates. Silymarin co-treatment showed marked protection in terms of morphology of the seminiferous tubules and the density of germ cells. Decomposition of NADPH was measured spectrophotometrically at 340nm (25C).
Ahmad KH, Habib S: Indigenous knowledge of some medicinal plants of Himalaya region, Dawarian Village, Neelum Valley, Azad Jammu and Kashmir, Pakistan.
Also uncertain is the molecular mechanism by which reductions in GSH and other antioxidant molecules in Leydig cells, and thus increased oxidative stress, elicit reduced sensitivity to LH and thus reduced steroid formation. CCl4 treatment resulted in rupturing, displacement and alteration in shapes of seminiferous tubules. & K. Krause) from Nigeria. Development of spermatozoa from spermatogonial stem cells is regulated by various hormones and this whole process in controlled by the hypothalamic-pituitary-testicular axis. Diminished level of CAT, POD, SOD, GPx, GST, GR and GSH while enhanced level of TBARS, nitrite and H2O2 in testes samples of male rats was reversed back to the control, dose dependently, by the co-administration of JDEE. Prakash KC. Glutathione is one of them.
2009;47:13939. Annals of Biochemistry. Administration of silymarin in combination with CCl4 ameliorated the toxic effect of CCl4 and increased the activity level of CAT, POD, SOD in testes samples and nonsignificant difference with the control was recorded.
Treatment of CCl4 causes hypogonadism and decrease the level of testosterone in serum of rat [3, 4]. Earlier we have reported in vitro antioxidant and DNA protection ability of the methanol extract and its derived fractions. Clin Nutr.
Differential distribution of glutathione and glutathione-related enzymes in rabbit kidney: possible implications in analgesic nephropathy.
The project was funded by the Department of Biochemistry Quaid-i-Azam University Islamabad Pakistan. To 50l of tissue homogenate, 25l of 1mM oxidized glutathione, 50l of 0.1mM NADPH, 50l of 0.5mM EDTA and 825l of 0.1M sodium phosphate buffer (pH7.6) were added. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article.
Nor did treatment of the cells with tert-butyl hydroperoxide (t-BuOOH, 2 hours), an oxidant.
International Journal of Life-Sciences and Scientific Research.
Various reports revealed that carbon tetrachloride (CCl4) a well-established hepatotoxin also causes injuries in testes [2, 3].
Comparative study of the assay of Artemia salina L. and the estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant extracts. Group III were treated (i.p.) 1982;126:1318.  activity of glutathione-S-transferase was based on the formation of conjugate between GSH and 1-chloro-2,4-dinitrobenzene (CDNB). It is a prostrate perennial herb with a thick terminal group of large flower heads and a rosette of long spreading lobed leaves with purple mid veins, root long, tuberous.
Medical treatment to improve sperm quality. Free radical scavenging capability of JDEE may be the reason in defensive impact against CCl4 harmfulness. Increase in the diameter of seminiferous tubules and increase in germinal thickness has been demonstrated in rat with pomegranate juice and alcoholic extract of Nigella sativa seeds [27, 28]. Nitrite concentration was calculated in the tissue samples by using standard curve of sodium nitrite. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1975;250(14):547580.
Al-Sa'aidi JAA, Al-Khuzai ALD, Al-Zobaydi NFH. Animals were screened for mortality and morbidity for 14days .
To cope with such reactive species and the clinical disorders, it is essential to obtain dietary antioxidants which counter measure the excessive generation of free radicals [3, 4]. For acute toxicity study 18 SpragueDawley male rats of good health were randomly divided into six groups (3 rats in each).
It is suggested that the enhanced level of testosterone with JDEE in male rats might be beneficial to increase libido and sexual function of human being.
JDEE co-treatment to rats ameliorated the toxic effects of CCl4 in testes samples. The experiment was performed according to the instruction. Protective influence of Ficus asperifolia Miq leaf extract on carbon tetrachloride (CCl4)-induced testicular toxicity in rats.
 method. Chemical and Process Engineering Research. Privacy These results suggest that, as in aging, exposure to an increasingly pro-oxidant environment, over time, can have a negative impact on Leydig cell steroidogenic function, and that, indeed, increases in oxidative stress contribute to or cause the reduced testosterone production that characterizes Leydig cells aging. With the help of the Griess reagent nitrite assay was carried out according to the method of Green et al. After collection, roots were shade dried till the complete removal of moisture and samples were made to mesh sized powder by using plant grinder and powder (5kg) was soaked in crude methanol (10L) for extraction for 72h. The extraction was repeated two times with above procedure.
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Male infertility is a major clinical problem affecting 30% of the world population . Testes were excised from each animal and placed in saline solution. The reaction mixture was prepared by the addition of 1ml of 0.28nM phenol red, 2.0ml of sample homogenate, 5.5nM dextrose, 0.05M phosphate buffer (pH7) and horseradish peroxidase (8.5units) and incubated for 60min at 37C. Most aging men and many young men have reduced circulating testosterone levels, referred to as hypogonadism. 2006;2:27. Reprod BioMed Online. Exposure of rats to CCl4 prompted significant reduction in CAT, POD, SOD and GPx profile, exhausted the GSH level and elevated TBARS in testes .
In this regard we have assessed the activity level of antioxidant enzymes and lipid peroxidation in testes samples whereas the concentration of testosterone in serum of rat. At 412nm absorbance was immediately read and the GSH activity was presented by M GSH/g tissue. The homogenate was treated with an equal volume of (100l) of 0.3M NaOH and 5% ZnSO4. Root of Jurenia dolomiaea is used traditionally in various disorders involving oxidative injuries i.e. Increase of glutathione, testosterone and antioxidant effects of, https://doi.org/10.1186/s12906-017-1718-z, http://creativecommons.org/licenses/by/4.0/, http://creativecommons.org/publicdomain/zero/1.0/.
Bars with different letters indicate significant difference (P<0.05). To determine the difference among various treatments one way analysis of variance was estimated by using the Statistix 8.1. Plants of J. dolomiaea were collected in 2011 from Nazar zera area of Kohistan, Pakistan.
The supernatant was collected which was used for further analyses. Animals were off feed but had open access to water 15h prior of test samples. Sensitivity of the kit is 0.2nmol/L 50nmol/L. (1) Control, (2) Vehicle control, (3) CCl4 treated control, (4) CCl4+Silymarin (200mg/kg), (5) CCl4+JDEE (200mg/kg), (6) CCl4+JDEE (400mg/kg), (7) JDEE (400mg/kg).
40 ; Microphotographs of testes histology of different groups after J. dolomiaea ethyl acetate fraction (JDEE) treatment (H & E staining). alone, CCl4 (I ml/kg; 1:10v/v in olive oil) alone, JDEE (200mg/kg, 400mg/kg) with CCl4, and silymarin (200mg/kg) with CCl4 on alternate days for 60days.
2006;12:70414. Roots are cooked with maize flour and used for the treatment of bone fractures. Similar histopathological alterations were recorded while assessing the defensive impact of various extracts of plants on testes against CCl4 induced toxicity [3, 4]. Oxidative Med Cell Longev.
Indian Journal of Biochemical Biophysics. Like many antioxidants, glutathione can play an important role in reducing inflammation in the body, particularly the lungs.
2022 BioMed Central Ltd unless otherwise stated. Co-treatment of silymarin and the JDEE (200mg/kg, 400mg/kg) lessened the toxic effects of CCl4 and reversed the level of these parameters towards the control group.
The method of Ohkawa et al. with 1ml/kg of CCl4 (1:10; v/v) after dissolving in olive oil and DMSO (1:1; v/v). This study makes use of rats and the experimental protocol for the use of animal was approved (Bch#0248) by the ethical board of Quaid-i-Azam University, Islamabad Pakistan. Some features of this site may not work without it. Protective role of glutathione and evidence for 3, 4-bromobenzene oxide as the hepatotoxic metabolite. Biochem Pharmacol. As yet, cause-effect relationships have not been established, however. The reaction mixture consisted of 100l of tissue homogenate, 10l of 100mM FeCl3, 100l of 100mM ascorbic acid and 290l of sodium phosphate buffer (pH7.4).
Ethnomedicine in Himalaya: a case study from Dolpa, Humla, Jumla and mustang districts of Nepal. By using molar extinction coefficient (6.22103/M/cm) activity of GPx was assessed and expressed as nM of NADPH oxidized/min/mg protein. The authors declare that they have no competing interests. Bars with different letters indicate significant difference (P<0.05). Glutathione-S-transferases: the first enzymatic step in mercapturic acid formation. Antioxidant and antibacterial activities of the J. dolomiaea plant have also been evaluated . Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Cite this article. Food Chem Toxicol. However, the administration of JDEE (400mg/kg) alone to rats significantly enhanced the level of testosterone in serum as compared to the control group.  was followed to measure GSH activity in the testes samples. Concentration of TBARS, H2O2 and nitrite was diminished, dose dependently, and at higher dose of JDEE (400mg/kg) co-administered with CCl4 similar level of these parameters to that of the control group was observed. Absorbance change of 0.01 as units/min defines one unit CAT activity . General behavior of animals was noted after 120min of treatment. Absorbance of the reaction mixture was recorded at 560nm. To test the effect of decreased GSH on steroid formation, we first cultured MA-10 Leydig cells with BSO short-term (24 hours). Pick A, Keisari Y. Superoxide anion and hydrogen peroxide production by chemically elicited peritoneal macrophages-induction by multiple nonphagocytic stimuli. 2010;9:18794. 1984;33(11):18017. In vivo evaluation of the plant was needed to ensure its protective effects against CCl4 induced toxicity in testes of rat. 1. Further, administration of JDEE (400mg/kg) alone to rats significantly (P<0.05) increased the level of GSH as compared to the control group. As JDEE interfere with the oxidation process by scavenging, reducing and chelation of free radical  that is reflected in decrease of TBARS and increase in total antioxidant capacity. Hypogonadism is estimated to affect about 5 million American men, including both aged and young. BMC Complementary and Alternative Medicine However, an admirable effect of JDEE alone to male rats was the enhanced level of GSH in testes and testosterone in serum samples.
Rats when orally treated with JDEE (400mg/kg) alone did not alter the activity level of these enzymes as compared to the control group. Treatment of JDEE (400mg/kg) alone to rats did not change the level of protein as compared to the control group. , NADPH was used as substrate in order to determine GPx activity.
Jurinea macrocephala Royle) has a place with Family Asteraceae (Compositae).
The designed protocol (Bch#248) was then approved by the Ethical Committee of Quaid-i-Azam University, Islamabad, Pakistan. The activity level of GST, GPx and GR in testes samples treated with CCl4 decreased (P<0.05) as comparison to the control group. We make it easy to stay informed about the latest trends in mens health to help you perform at the top of your game.
Protective effect of plants against the CCl4 induced testicular toxicity have been reported [3, 4]. with JDEE (200mg/kg; 400mg/kg, respectively).
2009;23:1238. Journal of Applied Pharmaceutical Science.
JDEE hold the constituents (flavonoids, terpenoids and saponins) which directly or indirectly ameliorate the oxidative harm to distinctive cells and organs.
However, when the GSH-depleted cells subsequently were exposed acutely to t-BuOOH, intracellular reactive oxygen species concentration was significantly increased, and this was accompanied by reductions in steroid production. Tissues after becoming clear were embedded in paraplast. Deterioration of seminiferous tubules and germ cells, interstitial in part was vanished and replaced by fibroblast and inflammatory cells were recorded in testes of CCl4 treated group of this study. The mixture was then centrifuged at 6400g for 20min to get the protein free supernatant.
1 filter was used and methanol was evaporated on a rotary evaporator at 40C under reduced pressure to get the viscous material and fractionated on escalating polarity basis. For this the reaction mixture contained 25l of tissue homogenate, 25l of 20mM guaiacol, 75l of 40mM H2O2 and 625l of 50mM potassium phosphate buffer (pH5.0). Protein measurement with Folin phenol reagent. The protocol of Pick and Keisari  was followed to assess the level of hydrogen peroxide (H2O2) in the testes samples. Trk G, Snmez M, Aydin M, Yce A, Gr S, Yksel M, Aksu EH, Aksoy H. Effects of pomegranate juice consumption on sperm quality, spermatogenic cell density, antioxidant activity and testosterone level in male rats. Food and water were given ad libitum. Higher testosterone level with pomegranate juice and alcoholic extract of Nigella sativa seeds has been determined in rat [27, 28]. MRK has made substantial contribution to designing, analyzing and drafting of the manuscript. The control group orally received 15% DMSO in olive oil. Department of Biosciences, COMSATS Institute of Information Technology, Islamabad, Pakistan, Department of Biochemistry, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan, You can also search for this author in
Briefly, 500l of testes supernatant was mixed with 500l of sulfosalicylic (4%) to carry out precipitation. Histopathological observation of testes samples endorsed the alterations induced with different treatments. Increase in GSH level with juice and methanol extract of pomegranate peel has been determined in rat . Ojo OA, Ojo AB, Ajiboye B, Fadaka A, Imiere OB, Adeyonu O, Olayide I. Additionally, it is recognized cordial and is given in puerperal fevers. The higher dose of JDEE (400mg/kg) produced similar protective effects (P>0.05) for these enzymes to that of the silymarin co-treated group. These results suggested the evaluation of JDEE for sexual behavior. 1951;193:26575. JDEE constituted significant amount of total flavonoids 807.07.2mg rutin equivalent/g of JDEE. Testes samples were investigated for antioxidant enzymes, biochemical markers and histopathology while the serum samples were analyzed for the testosterone level. 2016;6(06):03741. (1) Control, (2) Vehicle control, (3) CCl4 treated control, (4) CCl4+Silymarin (200mg/kg), (5) CCl4+JDEE (200mg/kg), (6) CCl4+JDEE (400mg/kg), (7) JDEE (400mg/kg). Dwivedi SD, Wagay SA.
Roots are also used by the local population for loose bowels and stomachache .
Presence of saponins in the JDEE might be responsible for the release of pituitary LH while the flavonoids might have been implicated in the synthesis of androgens. EXCLI J. Tissue homogenates were first centrifuged at 1500g for 10min and then for 15min at 10000g. From the supernatant obtained a volume of 150l was added to the reaction mixture containing 50l of 186M phenazine methosulphate and 600l of 0.052mM sodium pyrophosphate buffer (pH7.0). In another study use of pomegranate juice increased the GSH in testis of rat . Over time, at times after GSH was reduced, steroid synthesis decreased, reminiscent of natural aging of primary Leydig cells. At 340nm absorbance of the reaction mixture was recorded and with molar extinction coefficient of 9.6103/M/cm, GST activity was determined, expressed as nM CDNB conjugate formed/min/mg protein. alone with 400mg/kg of JDEE.
The concentration of testosterone in serum was estimated through Astra Biotech kit purchased from Immunotech Company. Earlier we have investigated in vitro antioxidant and DNA protective ability. Antioxidant potential, DNA protection, and HPLC-DAD analysis of neglected medicinal Jurinea dolomiaea roots.
Sprague Dawley male rats (42) were equally divided in to 7 groups: control, vehicle control, JDEE (400mg/kg; p.o.) On testis against oxidative stress of carbon tetrachloride in rat. The altered histopathological changes induced with CCl4 were also diminished with co-treatment of JDEE. SOD activity was determined by Kakkar et al. It is apparent from Fig.
Enhanced level of GSH, thickness of germinal layers in testes and testosterone in serum with JDEE (400mg/kg) treatment alone to rats demanded the evaluation of JDEE for sexual behavior. The reaction mixture was incubated for 1h at 37C in a shaking water bath at 37C.
Effect of different treatments of J. dolomiaea ethyl acetate fraction (JDEE) on (a) catalase (b) peroxidase (c) superoxide dismutase (d) glutathione-S-transferase (e) glutathione peroxidase (f) glutathione reductase in testes of rat.
 was used to measure the TBARS (thiobarbituric acid reactive substances) in testes samples. This treatment reduced intracellular levels of GSH somewhat but had no effect on progesterone production by these cells at this early time. 1974;11(3):15169. The results indicated that administration of CCl4 to rats cause toxicity and depleted the level of GSH in testes as compared to the control group. 2014;2014:726241.
However, the activity level of CAT, POD, SOD in testes of rat with JDEE (400mg/kg) treatment alone was not changed as compared to the control group. Given the abundance of GSH in Leydig cells, we hypothesized that the experimental depletion of GSH would result in an increasingly pro-oxidant intracellular environment, and that this would cause reduced steroidogenic function. J Ethnobiol Ethnomed. However, co-administration of JDEE at higher dose of 400mg/kg showed more protection in terms of nitrite content than the reference silymarin treated group.