ABSTRACT Introduction Neurogenic erectile dysfunction (NED) responds poorly to phosphodiesterase 5 (PDE5) inhibitors. Lycium barbarum polysaccharide (LBP) has broad-spectrum anti-inflammatory and anti-apoptotic effects, but whether it improves the erectile function of neuropathic ED through the above-mentioned mechanism is unclear. Objective We intend to explore the role of LBP in the treatment of NED. Methods A rat model of bilateral cavernous nerve injury (BCNI) was constructed by crushing the CN 5 mm distal to the major pelvic ganglion (MPG). Tweety-four BCNI rats were randomly divided into BCNI group, BCNI + LBP-LD (20 mg/kg/day) group and BCNI + LBP-HD (40 mg/kg/day) group, whereas another 8 age-matched rats formed the control group. Erectile function was evaluated by measuring intracavernosal pressure (ICP) and mean arterial pressure (MAP). The penile tissues, and MPG along with CN were collected from each group of rats for subsequent histological and molecular biological analysis. The rat Schwann cell line S16 was used for CCK-8 and wound healing assay, and the Schwann cell (SC) markers S100 and Pmp22, neuroinflammatory indicators IL-1β, TGF-β1, apoptosis-related proteins Caspase-3, Bax, Bcl-2 and nerve growth factor NGF were verified by PCR and WB. Results ICP/MAP indicated that the erectile function of rats in the BCNI model group was severely impaired, which was rescued in the BCNI+LBP-LD and BCNI+LBP-HD groups. Rats in the BCNI model group can highly express inflammatory indicators NF-kb, TNF-α and apoptosis-related proteins Bax, Caspase-3. TUNEL confirmed that the level of apoptosis in the penile tissue of BCNI rats increased. The above-mentioned penile inflammation and apoptosis changes can be inhibited by LBP. In addition, both immunofluorescence and WB confirmed that rats in the LBP-LD and LBP-HD groups highly expressed nNOS and down-regulated the expression of TH. In vitro experiments have shown that LBP can promote the proliferation and migration of SC, increase the expression of Pmp22 and NGF, and the inflammation and apoptosis-related proteins were down-regulated. Conclusion LBP can relieve the inflammation and apoptosis at the original site of nerve injury by promoting the proliferation and migration of SC, and secretion of NGF, thereby inhibiting the inflammation and apoptosis caused by hypoxia after denervation of penile tissue, and improving penile erection. Disclosure Work supported by industry: no.

ABSTRACT Introduction Acetyl-L-carnitine (ALCAR), an acetyl derivative of L-carnitine, has been proven to promote nerve repair and regeneration in a variety of neurodegenerative diseases. Neurogenic erectile dysfunction (NED) caused by CN injury is a common complication after radical prostatectomy, which lacks efficient curative treatments. Objective Although attempts have been made to apply ALCAR to treatment of peripheral neuropathy, the effects of ALCAR on NED remain unknown. Therefore, we intend to explore the potential role of ALCAR in the treatment of NED. Methods A rat model of bilateral cavernous nerve injury (BCNI) was established by crushing the CN 5 millimeters distal to the major pelvic ganglion (MPG). Twenty-four BCNI rats were randomly divided into BCNI group, BCNI+ALCAR-LD (50 mg/kg/day) group, and BCNI+ALCAR-HD (100 mg/kg/day) group, whereas the remaining eight age-matched rats were sham-operated as controls. Fourteen days after intraperitoneal injection of ALCAR, erectile function was evaluated by measuring intracavernosal pressure and mean arterial pressure. The penile tissues and CNs at the distal damage point were collected from each group of rats for subsequent histological and molecular biological analysis. Rat Schwann cell line S16 was used for CCK-8 and wound healing assay, and the expressions of SC markers Pmp22 and nerve growth factor (NGF) were verified by PCR. Results First, we found that the erectile function of rats in the BCNI model group was severely impaired, which was improved considerably in both BCNI+ALCAR-LD and BCNI+ALCAR-HD groups. Then, we observed that the penis was fibrotic after denervation in the BCNI group, which was manifested as decreased smooth muscle content and increased collagen content in the corpus cavernosum, and high expression of fibrosis indicators including TGF-beta, CTGF, and Smad 2/3. The above changes were alleviated after the administration of low and high-dose ALCAR. Meanwhile, our results also showed that after ALCAR treatment, the NO/cGMP pathway was promoted and the RhoA/ROCK pathway was inhibited in the corpus cavernosum of BCNI rats. Further, rats treated with ALCAR had high expression of ATF3 and S100 in the distal nerve tissues of the CN extrusion site, indicating the repairing effect of ALCAR on the CN. In vitro experiments showed that ALCAR promoted the proliferation and migration of SC, and increased the expression of Pmp22 and NGF. Conclusions ALCAR could boost CN repair and regeneration, thereby suppressing penile fibrosis and improving penile erection. ALCAR could inhibit penile fibrosis and improve penile erection by promoting proliferation and migration of SC and secretion of NGF to promote nerve repair and regeneration. Our study provides new insight into the therapeutic effect of ALCAR and preclinical evidence for a potential treatment strategy for NED. Disclosure Work supported by industry: no.

Purpose The pathogenesis of idiopathic hypogonadotropic hypogonadism (IHH) is genetically complex. The aims of this study were to investigate the genetic profile and clinical manifestation of IHH in a Chinese pedigree and to discover new IHH-associated genes. Methods The first step was to follow up the clinical phenotype and therapeutic outcomes of the pedigree in university hospital. The second step was that mutation screening was performed in this pedigree and 100 healthy controls. The third step was to further verify the pathogenicity of the discovered rare sequencing variant (RSV) by functional experiments. Whole exome sequencing, Sanger sequencing, testicular volume (TV), semen analysis, assessment of cell migration and necroptosis were performed. Results One heterozygous RSV (p.G517E) in CHL1 was identified in two male IHH patients and their mother in the pedigree, but not in healthy controls. All the three individuals exhibited olfactory impairment. hCG/hMG treatment significantly improved TV, serum testosterone and/or semen parameters of the two male patients. Functional analysis indicated that CHL1 significantly regulated GnRH neuronal cell line (GN11 cells) migration and necroptosis, with alteration of ERK1/2 activation, calcium loading, and transcription of RIPK3 and MLKL. However, the above processes were negatively influenced by the CHL1 RSV. Conclusions Our study reports the genetic relevance of CHL1 in IHH, and characterizes the phenotypic and therapeutic profiles in patients carrying the CHL1 RSV. CHL1 may act as a new IHH-associated gene, and should be taken into consideration in future investigations for this field.