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Objective:To observe the effects of four prostaglandin E2 (PGE2) receptors (EP 1-4R) on the activation of inflammasomes and cell damage in human retinal microvascular endothelial cells (hRMEC) in a high glucose environment. Methods:The hRMEC were divided into normal group and high glucose group, and they were cultured in Dulbecco modified Eagle medium containing 5.5 and 30.0 mmol/L glucose, respectively. Flow cytometry was used to observe the apoptosis rate of the high glucose group and the normal group; enzyme chain immunosorbent assay (ELISA) was used to detect the level of PGE2 in the culture supernatant of hRMEC cells. Western blot was used to detect the protein expression of cyclooxyganese (COX2) and EP 1-4R in hRMEC. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of EP 1-4R mRNA in hRMEC. After 72 h of culture, the cells in the high glucose group were divided into control group, PGE2 group, EP 1-4R agonist group, PGE2+EP 1-4R inhibitor group, and dimethylsulfoxide group. According to the group, each group was given the corresponding agonist or inhibitor to continue the culture for 24 h. QRT-PCR was used to detect the expression of nucleotide-binding oligomerization structure-like receptor protein (NLRP3) and pro-interleukin (IL)-1β mRNA in each group of cells. ELISA was used to detect the content of IL-1β and lactic dehydrogenase (LDH) in the cell culture supernatant. Western blot was used to detect the expression of cleaved Caspase-1 in each group of cells. At the same time, hRMEC in a high glucose environment was given IL-1β stimulation for 24 h, and the activity of LDH in the supernatant of the cell culture medium was detected. Results:The apoptotic rate, COX2 protein expression, and PGE2 protein content in hRMEC in the high glucose group were significantly higher than those in the normal group, and they were time-dependent. Compared with the normal group, the expression levels of EP 1R, EP 2R, EP 4R protein and mRNA in hRMEC in the high glucose group were higher than those in the normal group ( P<0.05). Compared with the control group, PGE2 group ( t=4.627, P<0.01), EP 1-4R agonist group ( t=3.889, 3.583, 2.445, 3.216; P<0.05) hRMEC NLRP3 mRNA expression level was significantly increased; the expression level of pro-IL-1β mRNA increased, however the difference was not statistically significant (PGE2 group: t=1.807, P>0.05; EP 1-4R agonist group: t=1.807, 1.477, 0.302, 1.926, P>0.05). Compared with the PGE2 group, the expression of NLRP3 mRNA in hRMEC in the PGE2+EP 2R inhibitor group was significantly reduced ( t=2.812, P<0.05); the expression of pro-IL-1β mRNA in hRMEC in the PGE2+EP 3R inhibitor group was significantly increased ( t=4.113, P<0.01). The protein content of IL-1β in the cell culture supernatant of the PGE2 group, EP 1R agonist group and EP 2R agonist group was significantly higher than that of the control group ( t=5.155, 4.136, 4.817; P<0.01). Compared with PGE2 group, the protein content of IL-1β in the cell culture supernatant of the PGE2+EP 2R inhibitor group and the PGE2+EP 4R inhibitor group were significantly lower than that of the PGE2 group ( t=1.964, 4.765; P<0.05). The expression of cleaved Caspase-1 in hRMEC in the PGE2 group and EP 2R agonist group was significantly higher than that in the control group ( t=5.332, 4.889; P<0.05). The expression of cleaved Caspase-1 in hRMEC in the PGE2+EP 2R inhibitor group was significantly lower than that of the PGE2 group ( t=6.699, P<0.01). The LDH activity in the cell culture supernatant of the PGE2 group and the EP 2R agonist group was significantly higher than that of the control group ( t=4.908, 4.225; P<0.05). The activity of LDH in the cell culture supernatant of the PGE2+EP 2R inhibitor group was significantly lower than that of the PGE2 group ( t=5.301, P<0.01). Compared with the control group, the LDH activity in the culture supernatant of hRMEC cells in the high glucose environment was significantly increased ( t=3.499, P<0.05). Conclusions:The four receptors of PGE2 can activate NLRP3 and its effector molecules to varying degrees. EP 2R mainly mediates hRMEC damage under high glucose environment.
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Objective To explore the relationship between HHIP gene with COPD patients in Xinjiang Uygur population. Methods DNA was extracted from peripheral blood samples. HHIP gene (rs1828591 and rs12504628)polymorphic loci was detected by iMLDR technique in 233 cases and 292 controls in Uygur. Results There was no significant difference in the genotype,allele frequencies distribution of 2 haplotypes of HHIP (rs1828591 and rs12504628)between the disease group and the control group(P > 0.05). There was no differ? ence in 2 haplotypes of HHIP gene between the disease group and the control group(P > 0.05). Rs1828591 and rs12504628 gene showed significance with predicted FEV1%(P < 0.05). Conclusion Rs1828591 and rs12504628 gene are related with predicted FEV1%.
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Objective To investigate the association between polymorphism of S1, S2 locus allele in ADAM 33 gene and chronic obstructive pulmonary disease (COPD) and lung function in Xinjiang Uygur population. Methods Blood sam?ples from 217 COPD patients and 218 healthy controls were collected. Samples of DNA was extracted, and S1, S2 single nu?cleotide polymorphism (ADAM 33) was detected by ABI SNaPshot SNP genotyping. Results There were no significant dif?ferences in the frequencies of S1 locus CC, CT, TT genotypes and C, T alleles between patient group and control group (P>0.05). There were no significant differences in the frequencies of S1 locus CC, CG, GG genotypes and C, G alleles between patient group and control group (P>0.05). In patient group, there were no significant differences in S1, S2 locus genotype and clinical indicators of lung function display, and in the FEV1%predicted and FEV1/FVC (P>0.05). Haplotype analysis showed that there were no significant differences in three kinds of haplotypes between patient group and control group ( P>0.05). Conclusion There is no significant difference in the polymorphism of S1, S2 locus allele in ADAM 33 gene and the susceptibility to COPD in Xinjiang Uygur population.
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Objective To investigate the risk factors and treatment of silicone oil glaucoma (SOG).Methods Ninety-five eyes of 93 patients who underwent pars plana vitrectomy and silicone oil tamponade were evaluated in this study. The lens was removed in 58 eyes in which intraocular lens (IOL) was implanted in 10 eyes, so 48 eyes were aphakic. Silicone oil tamponade time was ≤6 months in 32 eyes,and ＞6 months in 63 eyes. The follow-up time ranged from 2 to 25 months, with a mean of (9.5±5.1)months. The fundus and intraocular pressure (IOP) were evaluated at 1 week, 2 weeks and 1 month after surgery. The diagnosis of SOG was established if the one-month postoperative IOP ＞ 21 mm Hg (1 mm Hg=0.133 kPa), and primary and neovascular glaucoma were excluded. After the diagnosis of SOG, carteolol hydrochloride and brinzolamide solution were immediately applied to the eye, and intravenous mannitol infusion was performed. If the IOP still can not be controlled after 1 week of such treatment, silicone oil removal surgery will be performed. If removal of silicone oil can not control the IOP,trabeculectomy surgery will be performed. Results SOG occurred in 21 eyes (22.1%), including 5 phakic eyes (10.6% of 47 phakic eyes) and 16 aphakic eyes (33.3% of 48 aphakic eyes) , 3 eyes (9.4% of 32 eyes)with short tamponade time (≤6 months) and 18 eyes (28.6% of 63 eyes) with long tamponade time (＞6months). The average silicone oil tamponade time was (10.8±5.1) months. Emulsification of the silicone oil occurred in 17 eyes (81.0%). After silicone oil removed, IOP was controlled in 17 eyes (81.0%) within one week. Conclusions Aphakic eye and the duration of silicone oil tamponade are the risk factors of SOG.Emulsification of silicone oil is the main cause. Silicone oil removal is an effective way to treat SOG.
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Objective To evaluate the effects of inflammatory cytokines,including tumor necrosisfactor TNF-α and interleukins(IL-6 and IL-8),to the expression of pigment epithelium-derived factor(PEDF)in human retinal pigment epithelium(RPE)cells.Method Cuhured primary human RPE cellswere treated with 20,2,0.2,and 0.02 ng/ml of TNF-α,IL-6 and IL-8 separately.The levels of PEDFexpression were determined by Western blot of the supernant after 6,1 2,24 and 48 hours of culture.Results PEDF secretion of RPE cells was inhibited by TNF-α,IL-6 and IL-8 in a time-and dose-dependent fashion.Compared with the controls,the expression of PEDF decreased significantly in 0.02ng/ml and 6 hours group(F=7.14,P<0.05),2.00 ng/ml and 48 hours group(F=14.05,P<0.01),and 20.00 ng/ml and 24 hours group(F=11.53,P<0.01).TNF-α was the most strength inhibitor(F=14,P<0.01).Conclusion TNF-α,IL-6,and IL-8 could suppress the expression of PEDF in thecultured human RPE cells.