Günter Ross

Afferent nerves are involved in the febrile response to injection of LPS into artificial subcutaneous chambers in guinea pigs

In guinea pigs, fever was induced by injections of 100 or 10 microgram/kg lipopolysaccharide (LPS) into artificial subcutaneous chambers and analysed under the influence of the local anesthetic, ropivacaine (ROPI), which was administered into the chamber at a dose of 10 mg/kg 30 min prior to LPS. In response to injections of 100 microgram/kg LPS into the subcutaneous chambers, fever was not modified by pretreatment with ROPI. High amounts of bioactive tumor necrosis factor (TNF) alpha and interleukin-6 (IL-6) were measured in the lavage of the chambers after administration of LPS. Comparatively low concentrations of both cytokines (0.5-4% of the concentrations in the lavage fluid) were detected in blood plasma simultaneously. In response to injections of 10 microgram/kg LPS into the subcutaneous chambers, fever was significantly reduced by pretreatment with ROPI to about 60% of the febrile response of control animals. Levels of TNF and IL-6 were lower in response to the reduced dose of LPS. TNF in plasma was even below the limit of detection. The suppression of fever by the local anesthetic was not observed when ROPI was subcutaneously injected into the contralateral site of the chamber position so that a systemic effect of ROPI in the reduction of fever can be excluded. The results indicate a participation of afferent neural signals in the manifestation of fever. This effect becomes obvious only if the dose of the applied inflammatory stimulus (LPS) is not high enough to activate a systemic generalised inflammatory response.

Parathyroid hormone–related peptide improves contractile responsiveness of adult rat cardiomyocytes with depressed cell function irrespectively of oxidative inhibition

AbstractParathyroid hormone–related peptide (PTHrP) was found to improve contractile function of stunned myocardium in pigs. The peptide is released from coronary endothelial cells during ischemia and significantly improves post–ischemic recovery. The present study was aimed to decide whether such an induction of contractile responsiveness of the heart requires co–activation of adjacent cells or is a genuine phenomenon of cardiomyocytes. A second aim of this study was to decide whether such an improvement is linked to depressed cell function in general or oxidative inhibition. Isolated adult ventricular cardiomyocytes from rats were constantly paced at 0.5 Hz for 10 min. Cells were exposed to a brief oxidative inhibition by addition of 0.5 mmol/l potassium cyanide (KCN) in the presence of glucose. Under these conditions, cells stopped beating after 280 s on average. 30 s before they stopped to beat, cells had already developed a reduction in cell shortening, maximal relaxation and contraction velocity. In the co–presence of PTHrP (1–34) (100 nmol/l) cells continued to beat regular and did not develop reduced cell shortening, irrespectively of oxidative inhibition. In a second attempt, cells were exposed to the NO donor SNAP (100 µmol/l) or 8–bromocGMP (1 mmol/l). As expected both agents reduced cell shortening significantly. This reduction in cell shortening was attenuated in co–presence of PTHrP, too. Finally, we investigated the effect of PTHrP on cell shortening at different extracellular concentrations of calcium. Although, PTHrP increased intracellular calcium at 2 and 5 mmol/l extracellular calcium, respectively, it improved cell shortening only at 5 mmol/l extracellular calcium. Thus, the beneficial effect of PTHrP on cell shortening was independent from intracellular calcium but dependent on the steepness of the calcium gradient between intra– and extracellular calcium. In conclusion, our study shows that PTHrP is able to improve cell shortening rapidly and directly irrespectively of the reason for the reduced cell function. Improved electromechanical coupling rather than intracellular calcium handling seems to be the most important mechanism.

N-terminal parathyroid hormone-related peptide hyperpolarizes endothelial cells and causes a reduction of the coronary resistance of the rat heart via endothelial hyperpolarization.

Parathyroid hormone-related peptide (PTHrP) is known to be a strong vasorelaxant peptide. The mechanisms by which PTHrP reduces the coronary resistance of the rat heart have not been worked out but seem to be independent of the classical PTH/PTHrP receptor-mediated, cAMP-dependent effect. In this study we hypothesized that PTHrP reduces the coronary resistance of the rat heart via endothelial cell hyperpolarization. Isolated microvascular endothelial cells from rat heart were incubated with PTHrP(1-36), and changes in the membrane potential were recorded via DiBAC fluorescence. Cells exposed to PTHrP showed a hyperpolarization of approximately 7mV. In the isolated Langendorff preparation, PTHrP-dependent vasodilatation of l-nitro-arginine-exposed hearts was abolished under depolarizing conditions (high potassium). Denudation of the endothelial cell layer significantly impaired the vasodilatory effect of PTHrP. In the presence of H89 (a cAMP/protein kinase A pathway antagonist) and indomethacin (a cyclooxygenase inhibitor), PTHrP dilated the vessels. In conclusion, PTHrP exerted a nitric oxide-independent vasodilatory effect that depends on endothelial cell hyperpolarization.

Vasodilatory effect of tuberoinfundibular peptide (TIP39): requirement of receptor desensitization and its beneficial effect in the post-ischemic heart.

Tuberoinfundibular peptide of 39 residues (TIP39) is a member of the parathyroid hormone (PTH) family and a highly specific ligand of the PTH-receptor type 2 (PTH-2r). Recent studies have shown vasoactive properties of TIP39 in the kidney. This effect was stronger after desensitization of the parathyroid hormone-receptor type 1 (PTH-1r). The aims of our study were three-fold: (1) to investigate the influence of TIP39 on coronary resistance (CR), (2) to investigate a possible cross-talk between vascular PTH-receptors in the cardiovascular system, and (3) to investigate whether the endogenously released PTHrP during ischemia induces such a desensitizing effect. Experiments were performed on isolated rat hearts that were perfused with a constant pressure (Langendorff mode) and the coronary flow was determined. Under basal conditions, TIP39 showed no influences on CR. However, TIP39 reduced the CR by approximately 22% after pre-treatment of the hearts with a PTH-1r agonist. This TIP39 effect was abolished either by co-administration of a PTH-2r antagonist or by inhibition of nitric oxide (NO) formation. In an ischemia-reperfusion model endogenously released PTHrP desensitized the PTH-1r and pre-ischemic addition of TIP39 reduced post-ischemic CR by about 28%. Again, this effect was completely abolished in the presence of the PTH-2r antagonist or the PTH-1r-antagonist or by inhibition of NO formation. However, no effect was observed when TIP39 was washed-out prior to ischemia or if the treatment with TIP39 was restricted to the reperfusion. Furthermore, a pre-ischemic application of the NO-dependent vasorelaxant bradykinin provoked a similar effect on the post-ischemic CR than TIP39. In conclusion, a NO-dependent vasodilatory effect of TIP39 was demonstrated if the PTH-1r is desensitized by either exogenously applicated PTHrP peptides or endogenously released PTHrP.