Immunosuppressed Wistar Rats Research Articles

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Low-dose bitter leaf improves sperm quality disrupted in immunosuppressed Wistar rats: An experimental study.

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Oral Candidiasis in Immunosuppressed Wistar Rats (Rattus norvegicus) Post Dexamethasone Injection at 7.2 mg/kg and 16 mg/kg Doses

Objective: Oral candidiasis is an infection that occurs in the oral cavity and is caused by candida species, often Candida albicans. This infection commonly occurs in a condition of immunosuppression caused by dexamethasone. Due to the side effects of antifungal therapy, developing a standardized immunosuppressed animal model to induce oral candidiasis for new therapies is required. The aim of this study is to observe oral candidiasis in immunosuppressed Wistar rats post dexamethasone injection at 7.2 mg/kg and 16 mg/kg doses. Material and Methods: Twenty-one Wistar rats were divided into three groups: control group, treatment group 1 (injected with dexamethasone at a concentration of 7.2 mg/kg), and treatment group 2 (at a concentration of 16 mg/kg) for five days. Immunosuppression status was observed by leukocyte count and all the subjects’ palates were inoculated with C.albicans 0.1 ml of 15x108 UFC/ml 24 hours later. The subjects’ tongues were observed and confirmed by laboratory examination on day 10. A statistical analysis was performed using one way ANOVA, Kruskal–Wallis, Tukey HSD, and Mann-Whitney U tests. Results: A significant clinical appearance of the subjects’ tongues was observed only between C and T1 (p=0.023;p<0.05). Significant hyphal formation was observed between C and T1 (p= 0.037;p<0.05) and between C and T2 (p=0.007;p<0.05), and no significant difference was observed between T1 and T2. A significant increase in the colony count was also observed in similar results. Conclusion: Dexamethasone injection at doses of 7.2 mg/kg and 16 mg/kg is effective in triggering immunosuppression to induce oral candidiasis in immunosuppressed Wistar rats. Keywords Dexamethasone; Immunosuppression; Oral candidiasis.

Three Growth Factors Induce Proliferation and Differentiation of Neural Precursor Cells In Vitro and Support Cell-Transplantation after Spinal Cord Injury In Vivo.

Stem cell therapy with neural precursor cells (NPCs) has the potential to improve neuroregeneration after spinal cord injury (SCI). Unfortunately, survival and differentiation of transplanted NPCs in the injured spinal cord remains low. Growth factors have been successfully used to improve NPC transplantation in animal models, but their extensive application is associated with a relevant financial burden and might hinder translation of findings into the clinical practice. In our current study, we assessed the potential of a reduced number of growth factors in different combinations and concentrations to increase proliferation and differentiation of NPCs in vitro. After identifying a “cocktail” (EGF, bFGF, and PDGF-AA) that directed cell fate towards the oligodendroglial and neuronal lineage while reducing astrocytic differentiation, we translated our findings into an in vivo model of cervical clip contusion/compression SCI at the C6 level in immunosuppressed Wistar rats, combining NPC transplantation and intrathecal administration of the growth factors 10 days after injury. Eight weeks after SCI, we could observe surviving NPCs in the injured animals that had mostly differentiated into oligodendrocytes and oligodendrocytic precursors. Moreover, “Stride length” and “Average Speed” in the CatWalk gait analysis were significantly improved 8 weeks after SCI, representing beneficial effects on the functional recovery with NPC transplantation and the administration of the three growth factors. Nevertheless, no effects on the BBB scores could be observed over the course of the experiment and regeneration of descending tracts as well as posttraumatic myelination remained unchanged. However, reactive astrogliosis, as well as posttraumatic inflammation and apoptosis was significantly reduced after NPC transplantation and GF administration. Our data suggest that NPC transplantation is feasible with the use of only EGF, bFGF, and PDGF-AA as supporting growth factors.

Low dose bitter leaf improves sperm quality disrupted in immunosuppressed Wistar rats: An experimental study.

BackgroundSynthetic prednisolone (PRED) is a widely used over-the-counter glucocorticoid. Glucocorticoids have inhibitory effects on the immune system and are often used as immunosuppressive agents. Suppressed immunity may impact fertility via the hypothalamic-pituitary-adrenal axis. Bitter leaf (BL) has been shown to improve sperm parameters, but its effects on immunosuppression-associated infertility have not yet been documented.ObjectiveTo determine the fertility effects of bitter leaf on immunosuppressed Wistar rats.Materials and MethodsA total of 30 male adult Wistar rats were randomly assigned to 6 groups (n = 5/each). Group A served as a control and were given distilled water in addition to normal feeds, group B received 2 mg/kg PRED for 14 days and served as the standard immunosuppressed group, and groups C-F were immunosuppressed as in B but in addition received 50 mg/kg levamisole, low-dose (250 mg/kg) BL, high-dose (375 mg/kg) BL, and low-dose BL + levamisole, respectively. The CD4 counts, hematological parameters, and sperm parameters were analyzed and compared.ResultsThere were significant decreases in sperm motility, progressive motility, morphology, and life/death ratio in the animals given PRED only compared to the controls (p = 0.002, 0.001, 0.001, and 0.01, respectively). These were significantly increased in the treated groups, and animals given levamisole and 250 mg/kg BL showed significantly increased sperm counts compared to the controls (p = 0.04 and p = 0.04, respectively).ConclusionLow-dose BL (250 mg/kg) restored the sperm parameters altered by prednisolone administration.

The Effect of Hyperbaric Oxygen Therapy to the Amount of Lymphocytes in Oral Candidiasis Immunosuppressed Model

<p><strong><em> </em></strong></p><p><strong><em>Background: </em></strong><em>Oral candidiasis generally caused by Candida albicans with prevalence of 30-50%. C. Albicans proliferation in the oral cavity was canducted by decreasing immune system of the host. Lymphocytes activation in immune system happened when there is a contact between host cell with cell wall of C. albicans as a result of C. albicans antigen. One of therapy to suppress the Candida infection sistemically is by giving hyperbaric oxygen therapy. <strong>Purpose: </strong>To determine the effects of giving hyperbaric oxygen therapy to the amount of lymphocytes in oral candidiasis immunosuppressed models. <strong>Methods:</strong> This research was post test only control group design. We used 12 wistar rats which were divided into 3 groups(n=4/3): K-(normal/ healthy wistar rats), K+(oral candidiasis immunosuppressed wistar rats), P1(oral candidiasis immunosuppressed wistar rats which were given hyperbaric oxygen therapy 5 days). K+ and P1 groups were immunosuppressed by giving dexamethasone 0,5mg/day/rat orally for 14 days, added with tetrasiklin 1 mg/day/rat. On the 3rd day immunosuppression, the rats were inducted with C.albicans smeared in the dorsum linguae once every two days for 12 days. <strong>Results:</strong> The data were analyzed with one way Anova test and showed significant difference among groups (p<0,05). To show the difference between each groups we used LSD test and showed significant difference (p<0,05) between K+ compared with P1, and K+ compared with K-. <strong>Conclusion:</strong> Hyperbaric oxygen therapy gives effect to the amount of lymphocytes in oral candidiasis immunosuppressed models.</em></p><p><em> </em><strong><em>Keywords:</em></strong><em> Oral candidiasis, immunosuppressed, hyperbaric oxygen therapy, lymphocytes</em></p><p> <strong><em>Correspondence</em></strong><strong><em>: </em></strong><em>Agni Febrina Pargaputri</em><em>, Departmen</em><em>t of</em><em> Oral </em><em>Biology </em><em>F</em><em>aculty of</em><em> </em><em>Dentistry</em><em>, Jl. Arif Rahman Hakim 150, Surabaya, Indonesia. </em><em>Phone:</em><em> 031-5945864, fax: 031-5912191, Email: </em><em><span style="text-decoration: underline;">agni.febrina</span></em><em><span style="text-decoration: underline;">@hangtuah.ac.id</span></em></p><p> </p>

Alleviation of cyclophosphamide-induced immunosuppression in Wistar rats by onion lectin (Allium cepa agglutinin)

Ethnopharmacological relevanceIn various traditional medicines, onion has been classified as an immune-boosting food. Recent studies have claimed this property due to the presence of bioactive organosulfur compounds, prebiotic fructo-oligosaccharides and an immunomodulatory protein, lectin (Allium cepa agglutinin; ACA) (Prasanna and Venkatesh, 2015. Characterization of onion lectin (Allium cepa agglutinin) as an immunomodulatory protein inducing Th1-type immune response in vitro. Int. Immunopharmacol. vol. 26, pp. 304–313). Aim of the studyThe aim of this study was to evaluate the immunoprotective properties of ACA in normal and cyclophosphamide (CP; 100μg/kg)-induced immunosuppressed Wistar rats. Materials and methodsWistar rats were administrated different doses of ACA (1, 10, and 100μg) to respective groups in normal as well as immunosuppressed animals. The effect of ACA on the status of immune organs was assessed by examining the splenic and thymic indices, and histopathological changes. The biomarkers for humoral immunity (serum IgG and IgA levels) and serum pro-inflammatory markers (COX-2, TNF-α and IL-10) were measured by ELISA. ResultsACA showed immunoprotective properties by significantly promoting the restoration of lymphoid cell count by ~6 fold vs. model control (immunosuppressed animals) and promotes the immune response significantly (~1.5-fold) in CP-induced immunosuppressed animals compared to model control; production of pro-inflammatory molecules (COX-2 and nitric oxide) and expression levels of immune regulatory molecule (TNF-α) were elevated in a dose-dependent manner. ConclusionsThe observed in vivo results suggest that ACA has the potential to be used as a nutritional therapeutic to boost the immune status of immunosuppressed subjects brought about by CP administration.

Evaluation of biochemical and reproductive biomarkers in pre and during gestational period in immunosuppressed Wistar rats with mycophelonate sodium

Materials and methods The rats were divided into three groups: rats treated with water (CONT=10) rats orally treated with MFS (20mg/ Kg) daily for 15 days until a positive diagnosis of pregnancy (MICO-1=10) and rats orally treated with MFS (20mg/Kg) daily for 15 days prior to and during 21 days of pregnancy (MICO-2=10). It was evaluated on the 17th day of pregnancy the Oral Tolerance Glucose Test (OGTT) and on day 21 the rats were anesthetized and killed by decapitation and thus measured reproductive parameters (Index fertility, childbirth, pregnancy and number of live births and resorption) and serum biomarkers (lipid parameters and liver transaminases) Statistical significance was p <0.05. Results In pregnancy period, there were no changes in glucose data during OGTT evaluation, although MICO-2 rats presented higher glycemia compared to other groups (CONT and MICO-1), with 170mg/dL. Furthermore, the MICO-2 rats presented other negative effects, including dead rats and total resorption of fetuses. The biochemical data presented decreasing of rates of serum lipid (cholesterol and lipoprotein fractions, triglycerides and alanine aminotransferase).

Evaluation on physiological biomarkers pre and during gestational period in immunosuppressed Wistar rats with mycophenolate sodium

Materials and methods The rats were divided into three groups: rats treated with water (CONT=10) rats orally treated with MFS (20mg/ Kg) daily for 15 days until a positive diagnosis of pregnancy (MICO-1=10) and rats orally treated with MFS (20mg/Kg) daily for 15 days prior to and during 21 days of pregnancy (MICO-2=10). It was weekly evaluated body weight, blood glucose, and water and food intake (before and during pregnancy). Statistical significance was p <0.05.

Evaluation of biochemical and reproductive biomarkers on gestational period in immunosuppressed Wistar rats with Cyclosporin

Evaluation of biochemical and reproductive biomarkers on gestational period in immunosuppressed Wistar rats with Cyclosporin

Eugenol biologic activity in immunosuppressed rat females withCandida albicans genital infection: histocytological changes

From our prior studies concerning eugenol's biologic activity we observed the very good in vitro antifungal efficiency of this natural compound. Those positive results generated the need to supplement the available information with a comparative in vivo animal model. In this context, our current study was proposed to ascertain and compare the effects of eugenol with nystatin with a placebo control group (saline solution) by evaluating the cytohistological alterations in immunosuppressed Wistar female rats genitally infected with Candida albicans. The observed histological alterations were most apparent in the placebo group. Candidiasis affected the liver, ovaries, kidneys, and spleen of the infected female rats. When compared to the groups treated with eugenol and nystatin, these alterations were either less discernible or were not reported at all, except for the liver degenerations, which were attributed to the immunosuppressive activity of drugs used and not to the Candida infection. The efficiency in the in vivo experimental candidiasis exerted by eugenol upon the tissues' histoarchitecture allows us to seriously consider it as a promising antifungal active substance, useful therapeutically in genital candidiasis and, in our opinion, with comparable biologic activity to that of nystatin.

Hibiscus tiliaceus: A possible immunomodulatory agent

ObjectiveThe objective of the study is to evaluate the immunomodulatory effect of methanolic leaf extract of Hibiscus tiliaceus (MLHT) in pyrogallol induced immunosuppressed Wistar rats. MethodsThe methanolic extract of leaves of H. tiliaceus was administered orally at the dosage levels of 250 mg/kg/day and 500 mg/kg/day b.w in Wistar rats for 28 days. The assessment of immunomodulatory activity, humoral and cellular immunity was studied by hemagglutination antibody (HA) titer, delayed type hypersensitivity (DTH), Neutrophil adhesion test and carbon clearance test. In order to induce immunosuppression in rats pyrogallol (100 mg/kg/day, p.o.) is used and septilin syrup (1ml/100 g/day, p.o.) used as standard as it is immunostimulating agent. Hematological and biochemical were estimated by standard methods. ResultsOral administration of MLHT showed a significant increase in the production of circulating antibody titer in response to sheep red blood cells (SRBCs). A significant (P < 0.001) increase in both primary and secondary HA titer was observed when compared to control group, whereas in H. tiliaceus showed significant (P < 0.001) increase in HA titer. MLHT significantly (P < 0.001) potentiated the DTH reaction by facilitating the footpad thickness response to SRBCs in sensitized rats. Also MLHT evoked a significant (P < 0.001) increase in percentage neutrophil adhesion to nylon fibers and phagocytic activity. It also enhanced the production of RBC, WBC and hemoglobin. It does not much affect the biochemical parameters. ConclusionAn oral administration of the MLHT showed immunomodulatory effect in Wistar rats in a dose dependent manner. From the results obtained and reported phytochemical studies H. tiliaceus has a significant effect on both humoral and cellular immunity in experimental animals, this may be attributed to the polyphenols and flavonoid content of the plant extract.

Free radical scavenging capacity, anticandicidal effect of bioactive compounds from Sida Cordifolia L., in combination with nystatin and clotrimazole and their effect on specific immune response in rats

BackgroundInfectious diseases caused by fungi are still a major threat to public health, despite numerous efforts by researchers. Use of ethnopharmacological knowledge is one attractive way to reduce empiricism and enhance the probability of success in new drug-finding efforts. In this work, the total alkaloid compounds (AC) from Sida cordifolia L. (Malvaceae) have been investigated for their free radical scavenging capacity, antifungal and immunostimulatory properties.MethodThe antifungal activity was investigated against five candida strains using the microplate dilution method and the Fractional Inhibitory Concentration Index (FICI) of compounds was evaluated. The antioxidant activity of the samples was evaluate using three separate methods, at last, the immunostimulatory effect on immunosuppressed wistar rats was performed.ResultsAs for the antifungal activity, result varied according to microorganism. The results obtained in this antifungal activity were interesting and indicated a synergistic effect between alkaloid compounds and the antifungal references such as Nystatin and Clotrimazole. Antioxidant capacity noticed that the reduction capacity of DPPH radicals obtained the best result comparatively to the others methods of free radical scavenging. Our results showed a low immunostimulatory effect and this result could be explained by the lack of biologically active antioxidants such as polyphenol compounds lowly contained in the alkaloid compounds.ConclusionThe results of this study showed that alkaloid compounds in combination with antifungal references (Nystatin and Clotrimazole) exhibited antimicrobial effects against candida strains tested. The results supported the utilization of these plants in infectious diseases particularly in treatment of candida infections.

Cross-Linking Enhances Deposition of Human Endothelial Progenitor Cells in the Rat Heart after Intracoronary Transplantation

Transplantation of human endothelial progenitor cells (hEPCs) may improve vascularization and left ventricular function after myocardial infarction. The scope of this study was to explore, whether cross-linking of EPCs may enhance the deposition of cells in the rat heart after clinical-like, intracoronary transplantation. To this end, (111)In-oxinate-labeled hEPCs were infused by a minimally invasive technique into the coronary arteries of immunosuppressed Wistar rats under control conditions and after ischemia/reperfusion. In a second set of experiments hEPCs were treated with phytohemagglutinin to create small cell clusters prior to transplantation. Continous three-dimensional HiSPECT images for 1 h and after 48 h revealed that cell deposition was significantly higher when hEPCs were cross-linked. Therefore, cross-linking of hEPCs may provide a promising approach to enhance the number of trapped cells also in a clinical setting.

Nisin F in the treatment of respiratory tract infections caused byStaphylococcus aureus

To determine the antimicrobial activity of nisin F against Staphylococcus aureus in the respiratory tract. The respiratory tract of nonimmunosuppressed and immunosuppressed Wistar rats were colonized with 4 x 10(5) viable cells of S. aureus K and then treated by administering 8192 arbitrary units (AU) nisin F intranasal. Symptoms of pneumonia were detected in the trachea and lungs of immunosuppressed rats that had not been treated with nisin F. The trachea and lungs of immunosuppressed rats treated with nisin F were healthy. No significant differences were recorded in blood cell indices. The antimicrobial activity of low concentrations nisin F (80-320 AU ml(-1)) was slightly stimulated by lysozyme and lactoferrin. Nisin F inhibited the growth of S. aureus K in the respiratory tract of immunocompromised rats. Treatment with nisin F at 8192 AU proofed safe, as the trachea, lungs, bronchi and haematology of the rats appeared normal. Nisin F is nontoxic and may be used to control respiratory tract infections caused by S. aureus. This is, however, a preliminary study with an animal model and need to be confirmed with studies on humans.

Molecular typing of Pneumocystis jirovecii found in formalin-fixed paraffin-embedded lung tissue sections from sudden infant death victims.

Previous studies have provided histological evidence of an association between primary Pneumocystis infection and sudden infant death syndrome (SIDS). The aim of this work was to determine the species of clustered Pneumocystis organisms found in formalin-fixed paraffin-embedded (FFPE) lung tissue sections from Chilean sudden infant death (SID) victims. This approach needed first to optimize a DNA extraction method from such histological sections. For that purpose, the QIAamp DNA Isolation from Paraffin-Embedded Tissue method (Qiagen) was first tested on FFPE lung tissue sections of immunosuppressed Wistar rats inoculated with rat-derived PNEUMOCYSTIS: Successful DNA extraction was assessed by the amplification of a 346 bp fragment of the mitochondrial large subunit rRNA gene of the Pneumocystis species using a previously described PCR assay. PCR products were analysed by direct sequencing and sequences corresponding to Pneumocystis carinii were found in all the samples. This method was then applied to FFPE lung tissue sections from Chilean SID victims. Pneumocystis jirovecii was successfully identified in the three tested samples. In conclusion, an efficient protocol for isolating PCR-ready DNA from FFPE lung tissue sections was developed. It established that the Pneumocystis species found in the lungs of Chilean SID victims was P. jirovecii.

Radiosensitivity of different human tumor lines grown as xenografts determined from growth delay and survival data : Schwachofer JHM, Hoogenhout J, Kal HB, Koedam J, Van Wezel HPN. Department of Radiotherapy, University Hospital, P.O. Box 9101, 6500 HB Nijmegen. In Vivo 1990;4:253-8

Four human tumor lines were grown as xenografts in nude mice to determine whether xenografts derived from different types of tumors would show tumor-type dependent differences in response to single-dose irradiation, and whether these differences, paralleled clinical behavior. Xenografts from a neuroblastoma, a squamous cell carcinoma, a melanoma and a lung adenocarcinoma were studied in terms of growth delay and tumor control dose (TCD50). To exclude an immunoreaction of the host in the radiation response of the tumor xenografts, the tumor lines were tested for their growth in immunosuppressed Wistar rats. No differences in growth of xenografts in either immunodeficient mice or immunosuppressed rats were observed. Both growth delay and local tumor control as expressed by cure correlated well with clinical behavior of the tumor types of origin. This study demonstrates that radiosensitivity of different human tumor lines can be evaluated in terms of growth delay and tumor control dose50 when they are grown as xenografts. To exclude immune reactions, proper controls should be included. The sensitivities established from these evaluations parallel clinical behavior, thus offering a tool for analysis of human tumor radiosensitivity of histologically different tumor types.