ABSTRACT
@#Objective: To investigate the expression of Artemin in chondrosarcoma and its effect on proliferation and migration of endothelial cells, and to explore the mechanism. Methods: A total of 40 chondrosarcoma tissue samples (low degree (Ⅰ), 20 cases; high degree (Ⅱ,Ⅲ), 20 cases) surgically resected from patients, who were treated in Lianyungang HospitalAffiliated to Nanjing University of Chinese Medicine from May, 2015 to April, 2019, were collected for this study. Another 20 cases of normal cartilage tissue specimen from patients with amputations due to car accidents were served as control. The expressions of Artemin, vascular endothelial growth factor (VEGF), Ki-67 and CD31+ vascular density in tumor tissues were detected by immunohistochemistry.After being treated with 10 ng/ml Artemin, the changes of VEGF, stromal cell derived factor-1 (SDF-1), matric metalloproteinase 2 (MMP2) and MMP9 in the supernatant of SW1353 cell culture were detected by enzyme-linked immunosorbent assay (ELISA), and the effects of Artemin-treated chondrosarcoma cells on the migration and proliferation of ECV304 cells were detected by Transwell migration assay and MTT cell proliferation assay, respectively. Results: The expressions of Artemin and Ki-67 in the tissues of low-level group were significantly higher than those in the control group (all P<0.01); the expressions ofArtemin and Ki-67 in the tissues of high-level group were significantly higher than those in the low-level group (all P<0.01). The expression of Artemin was positively correlated with VEGF level and vascular density in chondrosarcoma tissues (all P<0.01);Artemin promoted the secretion of VEGF by chondrosarcoma cells, but had no significant effect on the secretion of SDF-1, MMP2 and MMP9. Artemin induced the proliferation and migration of ECV304 cells by promoting the secretion of VEGF by chondrosarcoma cells (all P<0.01). Conclusion: Artemin is highly expressed in chondrosarcoma tissues and has a positive correlation with the expression of VEGF and vascular density. Artemin can enhance the angiogenesis induced by chondrosarcoma.
ABSTRACT
In the present study, we investigated the role of peripheral ionotropic receptors in artemin-induced thermal hyperalgesia in the orofacial area. Male Sprague-Dawley rats weighting 230 to 280 g were used in the study. Under anesthesia, a polyethylene tube was implanted in the subcutaneous area of the vibrissa pad, which enabled drug-injection. After subcutaneous injection of artemin, changes in air-puff thresholds and head withdrawal latency time were evaluated. Subcutaneous injection of artemin (0.5 or 1 µg) produced significant thermal hyperalgesia in a dose-dependent manner. However, subcutaneous injection of artemin showed no effect on air-puff thresholds. IRTX (4 µg), a TRPV1 receptor antagonist, D-AP5 (40 or 80 µg), an NMDA receptor antagonist, or NBQX (20 or 40 µg), an AMPA receptor antagonist, was injected subcutaneously 10 min prior to the artemin injection. Pretreatment with IRTX and D-AP5 significantly inhibited the artemin-induced thermal hyperalgesia. In contrast, pretreatment with both doses of NBQX showed no effect on artemin-induced thermal hyperalgesia. Moreover, pretreatment with H-89, a PKA inhibitor, and chelerythrine, a PKC inhibitor, decreased the artemin-induced thermal hyperalgesia. These results suggested that artemin-induced thermal hyperalgesia is mediated by the sensitized peripheral TRPV1 and NMDA receptor via activation of protein kinases.
Subject(s)
Animals , Humans , Male , Rats , Anesthesia , Head , Hyperalgesia , Injections, Subcutaneous , N-Methylaspartate , Polyethylene , Protein Kinases , Rats, Sprague-Dawley , Receptors, AMPAABSTRACT
Objective To investigate artemin and its receptors GFRα3 expression in human pancreatic ductal adenocarcinoma and its significance. Methods Expression and distribution of artemin and GFRα3 in 100 cases of human pancreatic ductal adenocarcinoma(PDAC) and 40 cases of normal pancreatic tissue were detected by immunohistochemistry and RT-PCR, quantitative RT PCR. Relations of artemin and GFRα3 with clinicopathological characteristics, especially nerve invasion were analyzed. Results Nerve invasion was detected in 64 out of 100 cases of PDAC, and the rate of nerve invasion was 64%. The ratio of expression of Artemin, GFRα3 protein in PDAC/normal pancreatic tissue was 2.697 ± 0.231 and 2.599 ± 0.588; the ratio of expression of Artemin, GFRα3 mRNA was 7.01 and 4.63. Artemin and GFRα3 expression were associated with tumor location, differentiation degree, TNM staging, nerve invasion node metastasis, but it was not associated with age, sex and tumor size. Artemin and GFRα3 expression was more positively expressed in cancer cells close to nerve tissue than cells far from nerve tissue. Conclusions Artemin and GFRα3 were involved in the development of pancreatic cancer and their high expression was closely related to perineural invasion.
ABSTRACT
Background Glial cell line derived neurotrophic factor (GDNF) is determined to have a neurotrophy effect and promoting effect to the growth of axon.GDNF has been applied in ophthalmology.Research showed that artemin,a new member of GDNF family,has a better function in protection of neuron,but seldom relevant document of distruibution of artemin in retina is found so far.Objective The aim of the present study is to investigate the distribution and expression of artemin in normal rat retinal neuron cells and retinal ganglion cells,and imitate diabetic environment to observe the expression of artemin at the condition of high glucose.Methods Retinal tissue was isolated from clean neonatal SD rats and cultured by expand culture method in DMEM/F12 containing 10% fetal bovine serum.40 mmol/L of glucose was added in medium in the seventh day after culture for 12 hours as experimental group.The expression and location of artemin in retina were tested by real-time PCR and cell immunofluorescence assay.Use of experimental animals followed the Management Regulation of experimental animals of Jiangsu Province.Results Cultured cells showed the typical cell body and processes in the seventh day.Cultured retinal ganglion cells (RGCs) presented the red fluorescence for Thy1.1 antibody,and multiple fluorescence label revealed that RGCs exhibited the green fluorescence for artemin antibody and red fluorescence for Thy1.1 antibody,indicating artemin protein was positively expressed in cultured RGCs.The numbers of positive cells for Thy1.1 antibody was (442±9)/high field in normal culture group and (263±7) /high field in 40mmol/L glucose culture group,showing a significant difference between them (P＜0.05).The expression of artemin mRNA in normal culture group and in 40 mmol/L glucose culture group,was showing a considerably difference between them(P＜0.05).Conclusion Artemin can be expressed in cultured retinal neuron cells and RGCs in rats.High glucose environment down-regulate the expression of artemin.This study proved a new idea for protecting RGCs against damage.