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【Objective】 Tostudy the effect of sex-differentiated human IgG samples on Dendritic cells (DC) secretion of inflammation-related factors and explore the effect of residual sex hormones in IgG products (such as IVIg) on the secretion of IL-6 by DC. 【Methods】 According to the standard IVIg production process, the company was entrustedto prepare sex-differentiated plasma purified IgG samples, and two sex-differentiated IgG samples with different sex ratios (male to female ratio1: 0, 0: 1) were obtained. The samples and referenceswere treated with human DC (induced by THP-1 cells) respectively. After 24 h of culture, the chemokines (CCL2, CCL3, CCL4), adhesion molecule (ICAM-1) and inflammatory cytokines (IL-1, IL-6, IL-10, IL-12p70, IFN-) in the cell supernatant were detected, The effects of different samples on the secretion of inflammation-related factors by DC were compared. The effect of sex hormone residues on the anti-inflammatory ability of IgG products was preliminarily explored uing sex hormones and sex hormone receptor blockers. 【Results】 The samples in each group significantly inhibitedthe secretion of chemokines (CCL2, CCL3, CCL4) and the adhesion molecule (ICAM-1) by mature DC (compared with the PBS group, P<0.05), but significantly promoted the secretion of inflammatory cytokines (IL-1a/b, IL-6, IL-10, IL-12p70), compared with the PBS group(P<0.05). The results of sex hormone residues showed that therewere residues of estradiol (E2) and testosterone (TSTO) in sex-differentiated IgG samples and IVIg products. The experimental results of IVIg and sex hormone/sex hormone receptor blockers showed that residual E2 may promote the secretion of IL-6 by DC, which may be achieved through the E2 receptor ERb. 【Conclusion】 There are differences in the effect of IgG samples prepared from combined plasma with different sex ratios on the secretion of cytokines by DC, which may be related to the residual E2 in the products. The residual sex hormones in IVIg may promote the production and secretion of IL-6 through the sex hormone receptor ERb expressed in DC, and TSTO may have a collaboration effect to enhance the secretion-promoting effect of IL-6 by E2. This study provides a theoretical basis for whether sex hormone residues need to be considered in the quality control indicators of IVIg products.
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To investigate the mechanism of long non-coding RNA plasmacytoma variant translocation 1 (PVT1) in gastric cancer caused by (HP) infection. The expression of PVT1 was detected by quantitative real-time polymerase chain reaction in HP-infected normal gastric epithelial cells GES-1. Gastric cancer cell line SGC-7901 was transfected with PVT1 small interfering RNA and co-cultured with HP,and then the inflammatory cytokines such as tumor necrosis factor-α (TNF-α),interleukin (IL) -1β,IL-6 and IL-8 were detected. After PVT1 was knocked down,the effects of PVT1 on the proliferation and migration of gastric cancer cells were examined by cell scratch assay. RNA-pulldown combined with mass spectrometry was used to detect the protein binding to PVT1,and the result of mass spectrometry was verified by RNA-pulldown combined with Western blot. In HP-infected normal gastric epithelial cells GES-1,quantitative real-time polymerase chain reaction showed that PVT1 was significantly up-regulated (=7.160,=0.019). PVT1 was knocked down in gastric cancer cells,and then infected with HP. The expressions of inflammatory factors including TNF-α (=3.899,=0.011),IL-1β (=14.610,=0.000),and IL-8 (=6.557,=0.001) were significantly inhibited. Although PVT1 knockdown had no significant effect on the proliferation ability of gastric cancer cells,it inhibited the migration of cells. PVT1 might interact with RPS8 protein. PVT1 may act as a pro-inflammatory factor and regulate gastric cancer caused by HP infection.
Assuntos
Humanos , Linhagem Celular Tumoral , Movimento Celular , Citocinas , Metabolismo , Células Epiteliais , Biologia Celular , Microbiologia , Técnicas de Silenciamento de Genes , Infecções por Helicobacter , Patologia , Helicobacter pylori , Inflamação , RNA Longo não Codificante , GenéticaRESUMO
Objcctivc: To detect the mRNA expression leveb of inflammatory-related factors NLRP3, caspase-1. IL-lp. and 1L-18 in the murine macrophages infected by periodontitis patient's own tissue nucleic acid, and to discuss the effects of penodontitis patient' s own tissue nucleic acid on the inflammation-related factors in the macro. hacs. Methods: The mflammator. . eriodontal tissue sam. les were collected durin . eriodontal fla. surcr. of the chronic periodontitis patients, and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery. Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA. The cultured mouse macrophages RAW264. 7 were divided into control group and experiment group, then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA (the cDNA at a concentration of 1 mg • L ) were added into the RAW264. 7 cells, respectively. Real-time PCR was used to detect the mRNA expression levels of NLRP3 . Caspase-1. 1L-1(3. and 1L-18 in the macrophages in various groups at 4. 6 and 8 h after incubation. Rcsilts: The microscope observation showed that the mouse macrophages RAW264. 7 grew well with round and polygon shapes, clear cytoplasm, and full cell body. Compared with control group, the expression levels of NLRP3. Caspase-1. 1L-10. and 1L-18 mRNA in the RAW264. 7 cells in experiment group at 4. 6. and 8 h were increased significantly ( P<0. 05 or P<0. 01). and the expresaon levels of NLRP3. Caspase-1. 1L-10. and 1L-18 mRNA in RAW264.7 cells at 6 h in experiment group were the highest. Ccoclusico. The periodontitis patient's own tissue nucleic acids can promote the mRNA expressions of inflammation- related factors in the RAW264. 7 cells, suggesting that the periodontitis patient' s own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264. 7 cells.
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Objective:To detect the mRNA expression levels of inflammatory-related factors NLRP3, caspase-1, IL-1β, and IL-18in the murine macrophages infected by periodontitis patient's own tissue nucleic acid, and to discuss the effects of periodontitis patient's own tissue nucleic acid on the inflammation-related factors in the macrophages.Methods:The inflammatory periodontal tissue samples were collected during periodontal flap surgery of the chronic periodontitis patients, and the healthy periodontal tissue samples were collected from the patients without any periodontal diseases undergoing crown lengthening surgery.Then the total RNA from gingival tissue was extracted and reversely transcribed into cDNA.The cultured mouse macrophages RAW264.7were divided into control group and experiment group, then the healthy periodontal tissue cDNA and inflammatory periodontal tissue cDNA (the cDNA at a concentration of 1mg·L-1) were added into the RAW264.7cells, respectively.Real-time PCR was used to detect the mRNA expression levels of NLRP3, Caspase-1, IL-1β, and IL-18in the macrophages in various groups at 4, 6and 8hafter incubation.Results:The microscope observation showed that the mouse macrophages RAW264.7grew well with round and polygon shapes, clear cytoplasm, and full cell body.Compared with control group, the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18mRNA in the RAW264.7cells in experiment group at 4, 6, and 8hwere increased significantly (P＜0.05or P＜0.01) , and the expression levels of NLRP3, Caspase-1, IL-1β, and IL-18 mRNA in RAW264.7 cells at 6 hin experiment group were the highest.Conclusion:The periodontitis patient's own tissue nucleic acids can promote the mRNA expressions of inflammation-related factors in the RAW264.7cells, suggesting that the periodontitis patient's own tissue nucleic acid has an immunomodulatory effect on the activation of RAW264.7cells.
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Objective To study dynamically the correlation between endotheliocyte functions,inflammation-related factors and TCM syndromes in sepsis patients.Methods According to the TCM syndrome differentiation,68 septic patients were divided into Qi-fen group (23 cases),Ying-fen group (28 cases),and Xue-fen group (17 cases).The control group (26 cases) was built up for contrast.Serum von willebrand factor (vWf),nitric oxide (NO),tumor necrosis factor-? (TNF-?),interleukin 6 (IL-6),interleukin 2(IL-2),interleukin 4 (IL-4),and white blood cell (WBC) count of all groups were measured.Results TNF-?,IL-6,and WBC in all the sepsis groups were significantly higher than those in the control group (P