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Abstract Background The role of myeloperoxidase MPO is essential in lábujjak ízületeinek kezelésére szolgáló gyógyszerek killing of phagocytosed bacteria.

Certain steroid hormones increase MPO plasma concentration.

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Our aim was to test the effect of MPO, its inhibitor indomethacin, and certain steroid hormones on bactericidal activity. Methods Human polymorphonuclear leukocytes PMN were incubated with opsonised Escherichia coli and either MPO, indomethacin, estradiol, or hydrocortisone. Intracellular killing capacity was evaluated with UV microscopy after treatment with fluorescent dye.

Next, an in vivo experiment was performed with nine groups of rats: in the first phase of the study indomethacin treatment and Pasteurella multocida infection Iiindomethacin treatment without infection I0untreated control with does testosterone cause prostate problems Mi and untreated control without infection M0 ; in the second phase of the study rats with infection and testosterone treatment NTcastration, infection and testosterone treatment CTcastration, infection and estradiol treatment CEnon-castrated infected control N0and castrated infected control C0.

After treatment bacteria were reisolated from the liver and heart blood on agar plates, and laboratory parameters were analyzed.

Results Indomethacin did not have a remarkable effect on the bacterial killing of PMNs, while the other compounds increased bacterial killing to various degrees.

In the animal model indomethacin and infection caused ízületi fájdalom esetén melyik orvoshoz poor clinical state, a great number of reisolated bacteria, elevated white blood cell WBC count, decreased C-reactive protein CRP and serum albumin levels. Testosterone treatment resulted in less bacterial colony numbers in group NT, but not in group Könyök fájdalom alkar compared to respective controls N0, C0.

Estradiol treatment CE decreased colony numbers compared to control C0. Indomethacin treatment and castration weaken immune responses and clinical state of infected rats, while testosterone and estradiol have a beneficial effect. Background Does testosterone cause prostate problems radicals are very reactive compounds with one or more unpaired electrons on their outer orbitals. They are of great importance in the bacterial killing role of macrophages.

The most important reactive oxygen radical is the superoxide anion with limited reactivity, but it can be converted into other more reactive radicals.

HOCl deploys bactericidal activity on the bacteria ingested in the phagosome. Additionally, chloramines generated by HOCl reacting with phagosomal proteins also contribute to bacterial killing, but the full consequences of this are not yet clear [ 12 ].

The tissues are protected against free radicals by antioxidant enzymes such as SOD, catalase, glutathione reductase, α1-antitripsinpeptides with thiol groups glutathione, thioredoxin family and nutritional antioxidants vitamin C and E, carotenoids, trace elements [ 23 ]. Certain steroid hormones also play a role in antioxidant defence through various mechanisms. In our previous work we demonstrated that cortisol, ß-oestradiol E2progesterone, and testosterone decreased the superoxide release from human PMNs, while aldosterone and cortexolone, the precursor of cortisol did not have such property [ 4 ].

In the literature the most extensively studied steroid hormone is E2. It enhances the cellular anti-oxidative defence molecules, reduces the production of reactive oxygen species ROSactivates endothelial, inducible and neuronal nitric oxide synthase, and neutralizes the excess ROS in various cell types [ 5 ]. The mechanism of antioxidant action of hydrocortisone is scantly studied in the literature.

It is mainly referred to as the main biomarker of stress, which leads to oxidative changes [ 6 ].

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Other glucocorticoids have both glucocorticoid receptor mediated genomic and a rapid non-genomic antioxidant effect [ 89 ]. The antioxidant effect of testosterone is ambiguous in the literature. In one study it showed a receptor-mediated antioxidant effect in vitro on cerebellar granule cells [ 10 ], but according to others it caused a does testosterone cause prostate problems in the activity of antioxidant enzymes and led to lipid peroxidation in the prostate and testis of rodents [ 1112 ].

MPO has also been reported to have antioxidant capacity despite its known free radical producing activity. In fact, it has been found, that MPO release and activity in PMNs can be enhanced by certain steroids, like E2, testosterone, and prednisolone. Thus, the antioxidant effect of these steroid hormones might be carried out at least in part by the elevation of MPO activity and release [ 14 — 16 ].

On the other hand, the MPO inhibitor indomethacin, a non-steroidal anti-inflammatory drug, a nonselective cyclooxygenase inhibitor increases free radical production in PMNs [ 1417 ]. This could mean a beneficial effect in the bactericidal activity of PMNs. In our present studies we aimed to evaluate the effect of indomethacin on ex does testosterone cause prostate problems and in vivo bactericidal function.

We also tested MPO, E2, and hydrocortisone on antibacterial capability of human PMNs, and does testosterone cause prostate problems effect of indomethacin, testosterone, and E2 on an in vivo rat model.

Our ultimate aim was to evaluate if indomethacin can be useful as an adjuvant therapy in septic patients. Since the two experimental models are different, and sex steroids have various effects on many participants of the immune system besides PMNs, the comparison of the results of the two experiments needs caution.

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Methods Ex vivo effect of indomethacin, MPO, estradiol, and hydrocortisone on the bactericidal activity of human isolated PMNs Isolation of PMNs Venous blood was drawn from 6 male and 5 female donors, aged 23—40 years. All volunteers were non-smokers, took no medications, did not suffer from any known disease and consented to participation.

The blood was taken into EDTA tubes between 8—9 a. Ízületi fájdalom és izomfájdalom were separated within 2 hours of blood drawing with the following method. The blood was applied to Histopaque Sigma, —1 in layers for the sedimentation of red blood cells does testosterone cause prostate problems put aside for an hour.

Granulocytes thus separated were buffer-washed twice and centrifuged with × g. The bacterial strain was cultivated on blood agar plates overnight at 37°C. After adding 4 drops of antibody to the suspension it was incubated for 30 min at 37°C. The antibody was specifically produced for E.

Control cells were incubated without any treatment. After incubation, PMNs were mixed with 2× opsonised E. After having removed does testosterone cause prostate problems aliquots, the pellets were dyed with μl 1. The aliquots were removed and does testosterone cause prostate problems killing was examined in the pellets.

Acridine orange stains only dead bacteria, which fluoresce in orange when excited by ultraviolet light. Several fields of vision were examined for each sample in order to find the true amount of dead bacteria.

To express intracellular killing effect of the various drugs a scoring system from 0—5 was set up, where 0 indicates no bacterial killing and 5 indicates a hypothetical maximal killing. From the results of parallel measurements an average was calculated. Effect of indomethacin, testosterone, and estradiol on the bactericidal activity in vivo in rats The in vivo experiments were carried out in two phases. SPF adult male Wistar rats {Crl. Wi Br. In the first phase animals were kept in groups of eight, had access to conventional rat food Ssniff R-Z, Spezialdiaten GmbH, Soest, Germany and fresh drinking water ad libitum.

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The rats were given does testosterone cause prostate problems following treatments through a feeding tube for five days, once daily: groups M0 and Mi: 0. On the 3rd day of treatment rats in groups Mi and Ii were infected with Pasteurella multocida ssp. Prior to infection, Pasteurella multocida ssp. Single colonies were passed and cultivated again at 37°C for 24 hours.

Bacterium culture was washed with 2 ml saline. Rats were infected with a subcutaneous injection of 0. Uninfected animals got 0.

On the 5th day of treatment animals were anaesthetized with diethyl ether and exsanguinated through the abdominal aorta. Blood was collected for laboratory analysis of inflammatory markers and hormone levels.

Blood count, serum proteins, CRP, serum testosterone and estradiol were determined with well established automated laboratory methods used in clinical practice at the Szent Istvan University, Faculty of Veterinary Science, Department and Clinic of Internal Medicine, Budapest. Isolation of the infective agent was attempted from each liver and heart blood.

A loopful of sample was streaked onto Columbia blood agar CBA and plates were incubated at 37°C for 24 hours. The uninfected groups were negative for culturing while the majority of infected animals gave positive result with variable density. In the second in vivo phase 30 rats were used and all of the animals were infected and the effects of hormone treatments were examined.

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Animals were kept as described previously. Five groups of 6 were created, and 18 rats were castrated. After a day recovery period the following treatments were given for seven days, twice daily a.

On the 4th day of treatment all animals were infected with a subcutaneous injection of 0.

On the 7th day of treatment animals were anaesthetized and exsanguinated, laboratory and microbiological examinations were carried out as in the previous experiment.

Both experiments were approved by the university ethical committee. Statistical analysis The Statistica software was used for all analyses. Results Ex vivo effect of indomethacin, MPO, estradiol, and hydrocortisone on the bactericidal activity of human isolated PMNs The ex vivo effect of indomethacin, MPO, estradiol and hydrocortisone on bacterial killing are shown in Figure 1.

Due to technical problems not all experiments gave valuable results, thus the number of donors n is variable among treatment groups.

Indomethacin did not have a remarkable effect in either direction, using either scale. The three other compounds seem to increase bacterial killing of PMNs to various degrees: estradiol is the most efficient, while hydrocortisone has the least activity.

Results are expressed as percentage of control mean ± SD and as scores mean ± SD. For better visibility the scores were multiplied by 10 on the diagram. The error bars show standard deviation.

Int J Mol Sci. Published online Jan 8.

Full size image Effect of indomethacin, testosterone, and estradiol on the bactericidal activity in vivo in rats In the first phase of our in vivo studies 1—1 rats were wasted in the infected groups Mi, Iiand 2 in the uninfected group treated with indomethacin group I0 on the 4th day of treatment, i. Animals in groups I0 and Ii treated with indomethacin, without and with infection were weak and faint.

Ascites was found in two rats in group I0 and one in group Ii. After the reisolation of P. The greatest number of bacteria could be reisolated from the animals in group Ii, with confluent colonies does testosterone cause prostate problems 11 samples, vs 3 samples in group Mi. In group Ii none of the samples gave a negative reisolation, however, in three samples in group Mi no colonies were reisolated.