Sex hormone-like Effects of Icariin on T-cells immune modulation in spontaneously hypertensive rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Epimedium brevicornu Maxim as a Chinese herb, is recommended for the treatment of menopausal women with hypertension for 50 years. Icariin, as the main hydrophilic ingredient of Epimedium brevicornu Maxim, has been proven to be a plant sex hormone and lower blood pressure down. Here, we hypothesized that Icariin can regulate T cells differentiation which leads to the blood pressure decrease in castrated SHR rats. AIM OF THE STUDY: The present study aimed to investigate the effects of the exogenous estrogen, androgen and Icariin on T-cell modulation in hypertension. MATERIALS AND METHODS: Two weeks after castration, both male and female SHR rats were given estradiol, testosterone, and Icariin intervention respectively. Body weight, blood pressure, and heart rate were tested weekly. After six weeks, proportion of T helper cells (Th), cytotoxic T cells (Tc), and regulatory T cells (Tregs) in both peripheral blood mononuclear cells (PBMCs) and splenocytes were tested by flowcytometry. Serum levels of estrogen, testosterone, AngII, TNF-α, IL-17 were tested by Elisa. Aortic arches were isolated for HE and Masson staining. The expressions of ERß and AR in aorta were tested by Western-blot. RESULTS: In both male and female SHR rats, we found that Icariin and estradiol lower blood pressure, but testosterone elevates blood pressure. Similar as testosterone, Icariin can attenuate Tc and Th proportions and elevate Tregs proportion in both peripheral blood and splenocyte in male SHR, which can be blunt by flutamide. Besides, Icariin performs similar function as estradiol that attenuates Tc proportions and elevates Tregs proportion in both peripheral blood and splenocytes in female SHR, which leads to the lower blood pressure and can be partly blunt by fulvestrant. Testosterone increases AngII and TNF-α levels in serum, leading to the higher blood pressure in both male and female SHR rats. CONCLUSION: These results verified that Icariin, as a plant sex hormone, can regulate T cells differentiation related to blood pressure decrease in SHR rats.

Jiedu Huoxue decoction improves inflammation in rat type III prostatitis: The importance of the NF-κB signalling pathway.

This study aimed to investigate the mechanism of Jiedu Huoxue decoction (JDHXD) in type III prostatitis based on the NF-κB signalling pathway. Twenty-six Sprague-Dawley male rats were divided into blank control, model, positive (Prostate Plus), low-dose JDHXD, medium-dose JDHXD and high-dose JDHXD groups. Type III prostatitis rat model was established and confirmed with HE staining. NF-кB P50 and NF-κB P65 expression was detected with immunohistochemistry. NF-κB mRNA expression was detected with qRT-PCR. Protein expression of NF-κB and its inhibitor Iκ-Bα was detected with Western blot. Compared to the model group, a decrease in glandular hyperplasia and inflammation, and in NF-кB P50 and NF-κB P65 expression in the medium- and high-dose JDHXD groups was observed. NF-κB mRNA expression was significantly increased in the model group compared to control (p < 0.05), and significantly decreased in the JDHXD treatment groups compared to model group (p < 0.05). Protein expression of NF-κB was significantly increased in the model and low-dose JDHXD groups compared to control(p < 0.05), and significantly decreased in the medium- and high-dose JDHXD groups compared to model group (p < 0.05). Protein expression of Iκ-Bα was vice versa. JDHXD could be a potential treatment for type III prostatitis via its regulation of NF-κB and Iκ-Bα expression.

Testosterone Deficiency Causes Endothelial Dysfunction via Elevation of Asymmetric Dimethylarginine and Oxidative Stress in Castrated Rats.

BACKGROUND: Testosterone is believed to mediate the penile erectile response by producing adequate nitric oxide; therefore, testosterone deficiency results in erectile dysfunction through decreased nitric oxide bioavailability. However, the mechanisms underlying endothelial dysfunction in testosterone deficiency remain unclear. AIM: To investigate the mechanism of endothelial dysfunction in a rat model of testosterone deficiency. METHODS: Rats were distributed into 3 groups: castrated (Cast), castrated and supplemented with testosterone (Cast + T), and sham (Sham). In the Cast + T group, castrated rats were treated daily with subcutaneous testosterone (3 mg/kg daily) for 4 weeks; Sham and Cast rats received only the vehicle. OUTCOMES: Erectile function using intracavernosal pressure and mean arterial pressure measurements after electrical stimulation of the cavernous nerve, endothelial function using isometric tension, asymmetric dimethylarginine (ADMA) levels using ultra-performance liquid chromatography and tandem mass spectrometry, and inflammatory biomarker expression were performed 4 weeks after the operation. RESULTS: In the Cast group, the ratio of intracavernosal pressure to mean arterial pressure significantly decreased, acetylcholine-induced relaxation was lower, and serum ADMA, oxidative stress, and inflammation biomarker levels were significantly increased (P < .01). Testosterone injection significantly improved each of these parameters (P < .01). CLINICAL TRANSLATION: The present results provide scientific evidence of the effect of testosterone deficiency on erectile function and the effect of testosterone replacement therapy. STRENGTHS AND LIMITATIONS: This study provides evidence of the influence of testosterone deficiency on endothelial function by investigating ADMA and oxidative stress. A major limitation of this study is the lack of a direct link of increased ADMA by oxidative stress to inflammation. CONCLUSION: Testosterone deficiency increased not only ADMA levels but also oxidative stress and inflammation in castrated rats, which can cause damage to the corpus cavernosum, resulting in erectile dysfunction. Kataoka T, Hotta Y, Maeda Y, Kimura K. Testosterone Deficiency Causes Endothelial Dysfunction via Elevation of Asymmetric Dimethylarginine and Oxidative Stress in Castrated Rats. J Sex Med 2017;14:1540-1548.

Testosterone Replacement Therapy Prevents Alterations of Coronary Vascular Reactivity Caused by Hormone Deficiency Induced by Castration.

The present study aimed to determine the effects of chronic treatment with different doses of testosterone on endothelium-dependent coronary vascular reactivity in male rats. Adult male rats were divided into four experimental groups: control (SHAM), castrated (CAST), castrated and immediately treated subcutaneously with a physiological dose (0.5 mg/kg/day, PHYSIO group) or supraphysiological dose (2.5 mg/kg/day, SUPRA group) of testosterone for 15 days. Systolic blood pressure (SBP) was assessed at the end of treatment through tail plethysmography. After euthanasia, the heart was removed and coronary vascular reactivity was assessed using the Langendorff retrograde perfusion technique. A dose-response curve for bradykinin (BK) was constructed, followed by inhibition with 100 µM L-NAME, 2.8 µM indomethacin (INDO), L-NAME + INDO, or L-NAME + INDO + 0.75 µM clotrimazole (CLOT). We observed significant endothelium-dependent, BK-induced coronary vasodilation, which was abolished in the castrated group and restored in the PHYSIO and SUPRA groups. Furthermore, castration modulated the lipid and hormonal profiles and decreased body weight, and testosterone therapy restored all of these parameters. Our results revealed an increase in SBP in the SUPRA group. In addition, our data led us to conclude that physiological concentrations of testosterone may play a beneficial role in the cardiovascular system by maintaining an environment that is favourable for the activity of an endothelium-dependent vasodilator without increasing SBP.

The orexigenic effect of orexin-A revisited: dependence of an intact growth hormone axis.

Fifteen years ago orexins were identified as central regulators of energy homeostasis. Since then, that concept has evolved considerably and orexins are currently considered, besides orexigenic neuropeptides, key modulators of sleep-wake cycle and neuroendocrine function. Little is known, however, about the effect of the neuroendocrine milieu on orexins' effects on energy balance. We therefore investigated whether hypothalamic-pituitary axes have a role in the central orexigenic action of orexin A (OX-A) by centrally injecting hypophysectomized, adrenalectomized, gonadectomized (male and female), hypothyroid, and GH-deficient dwarf rats with OX-A. Our data showed that the orexigenic effect of OX-A is fully maintained in adrenalectomized and gonadectomized (females and males) rats, slightly reduced in hypothyroid rats, and totally abolished in hypophysectomized and dwarf rats when compared with their respective vehicle-treated controls. Of note, loss of the OX-A effect on feeding was associated with a blunted OX-A-induced increase in the expression of either neuropeptide Y or its putative regulator, the transcription factor cAMP response-element binding protein, as well as its phosphorylated form, in the arcuate nucleus of the hypothalamus of hypophysectomized and dwarf rats. Overall, this evidence suggests that the orexigenic action of OX-A depends on an intact GH axis and that this neuroendocrine feedback loop may be of interest in the understanding of orexins action on energy balance and GH deficiency.

[The effect of castration on bone in prostate cancer patients]. / Kastraation luustovaikutukset eturauhassyöpäpotilailla.

Surgical or chemical castration is widely used for the treatment of advanced prostate cancer. Common side effects of castration are e.g. sweating, increased body fat, decreased muscle mass, impotency and anemia. Castration increases the risk of osteoporosis and fractures in prostate cancer patients via hormonal effects to the bone. Castrated men should be recommended to increase physical activity. Calcium and vitamin-D supplementation is recommended. The European Association of Urology guidelines for prostate cancer recommend bone mineral density measurements before long-lasting castration therapy. Medical therapy should be considered for patients after low-energy fractures, after the diagnosis of osteoporosis based on bone mineral density measurements and for fracture high-risk patients based on clinical decision.

Antiandrogen monotherapy: indications and results.

Many patients with prostate cancer for whom hormonal therapy is indicated are still physically and sexually active; quality of life is therefore a vital issue when considering treatment options. Traditional castration-based therapies, although effective, have implications with respect to quality of life, causing loss of libido, impotence, fatigue, and reduced bone mineral density. Monotherapy with a nonsteroidal antiandrogen is an attractive therapeutic alternative to castration, offering effective therapy with potential quality-of-life benefits. Of the available nonsteroidal antiandrogens, bicalutamide has been most extensively evaluated in the monotherapy setting. Mature combined data (56% mortality) from 2 large randomized studies show no statistically significant difference in overall survival between bicalutamide 150-mg monotherapy and castration in patients with locally advanced, nonmetastatic (stage M0) disease. Survival outcome in patients with metastatic (stage M1) disease (43% mortality) favored castration, although the difference in median survival between the groups was only 6 weeks. Bicalutamide 150-mg monotherapy was associated with significant advantages compared with castration, in terms of sexual interest and physical capacity, in patients with either M0 and M1 stage disease. Data from a small subgroup of patients with stage M0 disease suggest that bicalutamide may also reduce the risk of osteoporosis compared with castration. Long-term therapy with bicalutamide 150-mg monotherapy is generally well tolerated, with a predictable side-effect profile. The most common side effects are male breast pain and gynecomastia. Emerging evidence also supports the use of bicalutamide 150 mg, both as immediate monotherapy and as adjuvant therapy in early stage (localized or locally advanced) prostate cancer.

Castration of calves: a study of methods used by farmers in the United Kingdom.

A postal survey of farmers was conducted to determine the main methods used to castrate calves, and by whom and how they were applied. Among the 28 per cent of farmers who replied, those who did castrate calves used one or more of three methods: the Burdizzo was used by 43 per cent of farmers, surgery by 39 per cent, and rubber rings by 32 per cent, with 10 per cent using more than one method. Calves were castrated at all ages from less than one week to over six months, with one third of them being castrated at an age that legally requires the operation to be done under local anaesthesia by a veterinary surgeon. Rubber rings were never used by veterinary surgeons, but they carried out 43 per cent of surgical castrations, which was the method of choice in older calves. Local anaesthetic was used on 15 per cent of farms, mainly for surgical castrations. Sixty-seven per cent of farmers using the Burdizzo applied it twice, with the majority correctly applying the second crush below the first, and 90 per cent used precautions to control infection after surgical castration.

Determination of prostatic secretion in rats: effect of neurotransmitters and testosterone.

To study the neuronal and hormonal control of prostatic secretion, the prostatic urethra was cannulated in urethane anesthetized rats. The volume of prostatic secretion was measured following infusion of cholinergic and adrenergic agonists intact animals. Prostatic secretion was elicited by norepinephrine, phenylephrine and carbachol; but not by clonidine, isoproterenol, pilocarpine, or acetylcholine. Phenylephrine and norepinephrine infusions caused a high initial rate of secretion, which then declined rapidly. Carbachol infusion, in contrast, produced a low but constant rate of secretion that was maintained for up to 1 hour. Histological examination of the prostate revealed contraction of smooth muscle surrounding prostatic ducts after infusion of phenylephrine and norepinephrine, but not carbachol. Prostatic secretion was also measured in castrated rats supplemented with various doses of testosterone. Testosterone exerted a dose dependent control of prostatic weight and secretory volume. These results indicate 1) alpha 1 receptor agonists can cause contraction of smooth muscle to expel fluid from the rat prostate, 2) carbachol induces prostatic secretion through a mechanism other than contraction of gland, and 3) testosterone has a primary role in controlling prostatic size.

[Effect of elcatonin on experimental osteoporosis induced by ovariectomy and low calcium diet in beagles].

Effect of elcatonin, a synthesized analogue of eel calcitonin, on experimental osteoporosis in beagles induced by ovariectomy and a low calcium diet was studied. Twelve female beagles, 15-18 months old, were divided equally into 3 groups. The first group of beagles were ovariectomized and then fed a low calcium diet, Ca: 0.08%, (ovariectomy plus low Ca diet group); the second group was subjected to the same procedure and diet plus daily injection of elcatonin, 1.0 u/kg, i.m., (elcatonin group); and the third group was fed a standard diet, Ca: 1.36%, (control group) for 6 months. The undecalcified bone sections from the iliac crest were analyzed using histomorphometric methods following in vivo double labeling with Calcein. In the ovariectomy plus low Ca diet group, there were a decrease in trabecular bone volume and an increase in both resorption and formation surface of the trabeculae compared with the control group. These changes of trabecular bone volume and resorption surface were significantly prevented by daily injection of elcatonin. The increase of formation surface of trabeculae and mineral appositional rate was noted in the elcatonin group in comparison with the ovariectomy plus low Ca diet group. These results suggested that elcatonin could prevent bone loss caused by deficiency of sex hormone and dietary calcium.

Dietary NaCl loads promote calciuria and bone loss in adult oophorectomized rats consuming a low calcium diet.

This study was undertaken to determine whether dietary supplements of NaCl would exaggerate osteopenia in oophorectomized (OOPX) rats consuming a low calcium (0.01% Ca) diet. Thirty 300 g OOPX rats with 45Ca-labeled bones were studied. Animals in group 1 were killed at the start of the experiment, whereas those in groups 2 and 3 were fed a low calcium diet for 2 months. Group 3 rats received NaCl (8 g/100 g diet). Salt increased the urinary excretion of sodium, calcium, phosphate, cyclic AMP, 45Ca and hydroxyproline but did not augment fecal excretion of calcium or 45Ca. Salt caused bone loss. The femora of NaCl-treated rats contained less 45Ca, less calcium, less phosphate and less mineral ash than those of rats killed at the start of the experiment. It is suggested that in OOPX rats consuming a low calcium diet, increased NaCl intake causes decreased renal tubular reabsorption of calcium and consequently lowered plasma Ca++. This results in stimulation of parathyroid hormone secretion and thus increased bone resorption. We conclude that NaCl supplements exacerbate osteopenia in adult OOPX rats consuming a low calcium diet. The effects of high dietary salt intakes on bone loss in postmenopausal women deserve further study.

Effects of castration and testosterone substitution on body composition and muscle metabolism in rats.

Male weanling Wistar rats were castrated or sham-operated and followed for 12 weeks without substitution or with large (2 mg . 14 days-1) or small (0.2 mg . 14 days-1) intramuscular dose of testosterone enantate. Castration without substitution was associated with lower body weight and smaller fat cell sizes in different adipose tissue depots. The epididymal and caudal subcutaneous depots were the most sensitive to castration. The percentage of fast-twitch high-oxidative (type II A) muscle fibers decreased in the non-substituted castrated animals. There was a decrease in phosphorylase and lactate dehydrogenase activities in the white portion of the gastrocnemius muscle of the castrates. These changes were reversed by the large dose of testosterone. Removal of testosterone by castration thus seems to "feminize" male rats with respect to body composition and muscle metabolism.

[Experimental studies on the castration osteoporosis in the rat. II. Behavior of bone minerals]. / Experimentelle Untersuchungen an der Kastrations-Osteoporose der Ratte. II. Verhalten der Knochenmineralien.

In Wistar rats the bone mineral content of calcium magnesium, phosphor, sodium and potassium was investigated 10 months after complete ovarectomy. 20 of ovarectomized animals obtained in this time 0,65 mg ethinyloestradiolsulfonate/kg/month orally. The ovarectomy decreased significantly the content of calcium and magnesium of femur, humerus, vertebra thoracis and lumbalis. After therapy with the oestrogen these minerals were increased, but not significantly. These results did not correlate with the degree of histomorphometrically established osteoporosis. Phosphor, sodium and potassium were not changed significantly. The role of oestrogen dosis was discussed.

Osteoporosis, oestrogens and oöphorectomy. A proposed new test of oestrogenic potency.

The role of oestrogens, endogenous and exogenous, in the pathogenesis of osteoporis is briefly reviewed. Aspects of research into effects of oestrogens on plasma calcium and phosphorus metabolism, conducted at Groote Schuur Hospital, are incorporated in the discussion. The current role of oestrogen therapy in the prevention of osteoporosis is presented. Evidence suggests that different oestrogenic substances produce different metabolic effects. On this basis a new test for assessment of oestrogenic potency is proposed. It is concluded that appropriate or specific oestrogens are of value in the prevention of osteoporosis, but play little or no role once the process is fully developed. A positive preventive measure recommended is conservatism with regard to ovaries during gynaecological operation on young women.

Oestrogen replacement therapy for prevention of osteoporosis after oophorectomy.

The value of oestrogen therapy in the prevention of osteoporosis after oophorectomy was assessed in 114 middle-aged women who participated in a double-blind controlled trial of mestranol in an average daily dose of 23 mug. The skeletal response to treatment was measured by a photon absorption technique. Where treatment was started within two months of operation subsequent bone mineral loss was prevented. Treatment started three years after oophorectomy caused a highly significant increase in bone mineral content. When treatment was delayed for six years mestranol failed to prevent subsequent bone mineral loss with age. These effects occurred independently of the associated humoral changes in calcium and phosphorus homoeostasis. Mestranol in this dosage appeared to be relatively safe, but it is too early to evaluate the long-term hazards of such therapy.