OBJECTIVETo investigate the effects of TNF-alpha, TNF-beta and the acceptor expression about mechanical renal trauma with extraneous ADM.

METHODSThere were 104 healthy adult plain grade Wistar rat, randomly divided into four groups:8 in the group of control, 32 in the group of trauma, 32 in the group injected ADM before trauma, 32 in the group injected ADM post trauma. The experimental model of rat kidney with mechanical trauma was prepared by striking the area of rat skin reflecting by kidney with free dropping ferrous hammer in the last three groups. ADM (0.1 nmol/kg) administrated by intraperitoneal injection at 10 minutes before trauma or post trauma respectively in injected groups. All rats were executed by drawing-out all the blood in their hearts. Renal tissue was investigated to study positive expression of TNF-alpha, TNF-beta, TNFR after SABC stained.

RESULTSTNF-alpha expression:the TNF-alpha expression of trauma group was more positive than it of control group in the wound early time. The expression of group injected post trauma was less than it of trauma group at 1 h (P < 0.01). The expression of group injected before trauma was less than it of trauma group at 6 h (P < 0.05) TNF-beta expression: the TNF-beta expression of trauma group was less than it of control group at 1 h and 6 h (P < 0.05). The TNF-beta expression of group injected post trauma was more positive than it of trauma group at the same time of 1 h and 6 h (P < 0.01). TNFR expression: the TNFR expression of trauma group was less than it of control group at 6 h (P < 0.01). The TNFR expression of group injected before trauma was more positive than it of trauma group in the at the same time of 1 h and 6 h (P < 0.01).

CONCLUSIONSThe TNFR can regulate the TNF-alpha and the TNF-beta in dynamic balancing. The regulation of TNFR is main to TNF-alpha. What the TNF-beta participated in renal trauma mainly is the anti-damage process. ADM can reduce the expression of TNF-alpha. ADM increases the expression of TNF-beta and TNFR.

Adrenomedullin ; pharmacology ; Animals ; Disease Models, Animal ; Female ; Kidney ; injuries ; metabolism ; Lymphotoxin-alpha ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Cell Line ; Glucose ; metabolism ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Inflammation underlies a wide variety of physiological and pathological processes, and plays a pivotal role in controlling pathogen infection. C1q/tumor necrosis factor (TNF) related proteins (CTRPs), a newly discovered adipokine family with conservative structure and wide distribution, has attracted increasing attention. The CTRP family consists of more than 15 members which fall into the characteristic C1q domain. Increasing studies have demonstrated that CTRPs are involved in the onset and development of inflammation and metabolism as well as related diseases, including myocardial infarction, sepsis and tumors. Here, we first clarified the characteristic domains of CTRPs, and then elucidated their roles in inflammatory-related diseases. Taken together, the information presented here provides new perspectives for therapeutic strategies to improve inflammatory and metabolic abnormalities.
Humans ; Complement C1q/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Inflammation/metabolism* ; Myocardial Infarction

Cell Line ; Humans ; Lipid Metabolism ; Neurons ; metabolism ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo compare the ability of the polysaccharide from various Porphyromonas gingivalis (Pg) type and clinical strains in inducing THP-1 cells to produce cytokines interleukin(IL)-1β, IL-8, and tumor necrosis factor(TNF)-&alpha;, in order to analyze the immunogenicity of Pg polysaccharide components and the virulence-associated factors of this periodontal pathogen.

METHODSThe bacterial polysaccharide was extracted from high virulent Pg strains, W83, SJD2, SJD12 and low virulent Pg, ATCC33277, SJD4, SJD5, and SJD11 by phenol-water extraction. The extracted polysaccharide was used to stimulate the THP-1 cells with different simulation periods and doses. The level of the cytokines, including IL-1β,IL-8 and TNF-&alpha; in the cell culture suspension was measured by enzyme-linked immunosorbent assay(ELISA).

RESULTSThe polysaccharide extraction of Pg strains was composed of lipopolysaccharide(LPS) and capsular polysaccharide. The secretion of IL-1β, IL-8 and TNF-&alpha;, produced by the THP-1 cells showed in a time- and dose-dependent manner in the medium containing 10% fetal bovine serum. The level of these cytokines of the high virulent strains was higher than that of the low virulent strains in medium containing 1% fetal bovine serum.Four hours after stimulation with polysaccharide extracted from high virulent strains, the levels of IL-1β,IL-8, and TNF-&alpha; in the cell suspension were (1 639 ± 497), (1 648 ± 513) and (140 ± 48) µg/L, respectively, whereas for low virulent strains, the levels of IL-1β, IL-8, and TNF-&alpha; were (773 ± 382), (892 ± 400) and (67 ± 33) µg/L, respectively.

CONCLUSIONSPolysaccharide extracted from Pg could induced the THP-1 cells to secrete the cytokines of IL-1β, IL-8 and TNF-&alpha;. The level of the cytokines produced by the THP-1 cells associates with the bacterial virulent properties.

Cytokines ; metabolism ; Interleukin-1beta ; Interleukin-8 ; Lipopolysaccharides ; physiology ; Porphyromonas gingivalis ; metabolism ; Tumor Necrosis Factor-alpha

OBJECTIVE: To investigate the role of acetylated modification induced by coactivator p300 in lipopolysaccharide (LPS)- induced inflammatory mediator synthesis and its molecular mechanism. METHODS: Agilent SurePrint G3 Mouse Gene Expression V2 microarray chip and Western blotting were used to screen the molecules whose expression levels in mouse macrophages (RAW246.7) were correlated with the stimulation intensity of LPS. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (chip-qPCR) were used to verify the binding of the molecules to the promoters of IL-6 and TNF-α genes. The effects of transfection of RAW246.7 cells with overexpression or interfering plasmids on IL-6 and TNF-α synthesis were evaluated with ELISA, and the binding level of the target molecules and acetylation level of H3K27 in the promoter region of IL-6 and TNF-α genes were analyzed by chromatin immunoprecipitation sequencing technique (chip-seq). RESULTS: Gene microarray chip data and Western blotting both confirmed a strong correlation of p300 expression with the stimulation intensity of LPS. Immunocoprecipitation confirmed the binding between p300 and c-myb. The results of EMSA demonstrated that c-myb (P < 0.05), but not p300, could directly bind to the promoter region of IL-6 and TNF-α genes; p300 could bind to the promoters only in the presence of c-myb (P < 0.05). The expressions of p65, p300 and c-myb did not show interactions. Both p300 overexpression and LPS stimulation could increase the level of promoter-binding p300 and H3K27 acetylation level, thus promoting p65 binding and inflammatory gene transcription; such effects were obviously suppressed by interference of c-myb expression (P < 0.05). Interference of p65 resulted in inhibition of p65 binding to the promoters and gene transcription (P < 0.05) without affecting p300 binding or H3K27 acetylation level. CONCLUSION LPS can stimulate the synthesis of p300, whose binding to the promoter region of inflammatory genes via c-myb facilitates the cohesion of p65 by inducing H3K27 acetylation, thus promoting the expression of the inflammatory genes.
Acetylation ; Animals ; Inflammation Mediators ; Interleukin-6/metabolism* ; Lipopolysaccharides/pharmacology* ; Mice ; Tumor Necrosis Factor-alpha/metabolism*

OBJECTIVE: To investigate the effects of Tongxinluo (TXL) on thromboangiitis obliterans (TAO) and the underlying mechanisms. METHODS: Ninety male C57/BL6J mice were randomly divided into 6 groups according to a random number table: the sham group, TAO model group, Compound Danshen Tablet (CDT) group, and the high-, medium-, and low-dose TXL groups. All mice except the sham group were injected with sodium laurate (0.1 mL, 5 mg/mL) in the femoral artery to establish TAO mouse model. After modeling, mice in the sham and TAO model groups were intragastrically administered 0.5% (w/v) sodium carboxymethylcellulose, mice in the CDT group were intragastrically administered 0.52 g/kg CDT, and mice in the TXL-H, TXL-M, and TXL-L groups were intragastrically administered 1.5, 0.75, and 0.38 g/kg TXL, respectively. After 4 weeks of gavage, the recovery of blood flow in the lower limbs of mice was detected by Laser Doppler Imaging. The pathological changes and thrombosis of the femoral artery were observed by morphological examination. The expressions of tumor necrosis factor α (TNF-α) and inducible nitric oxide synthase (iNOS) in the femoral artery wall were detected by HE staining. Levels of thromboxane B2 (TXB2), 6-keto-prostaglandin F1α (6-keto-PGF1α), endothelin-1 (ET-1), interleukin (IL)-1β and IL-6 were measured using enzyme-linked immunosorbent assay (ELISA). Levels of activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (FIB) were detected by a fully automated biochemical analyzer. RESULTS: TXL promoted the restoration of blood flow in the lower limbs, reduced the area of thrombosis in the femoral artery, and alleviated the pathological changes in the femoral artery wall. Moreover, the levels of TXB2, ET-1, IL-6, IL-1β, TNF-α and iNOS were significantly lower in the TXL groups compared with the model group (P<0.05 or P<0.01), while the level of 6-keto-PGF1α was significantly higher (P<0.01). In addition, APTT, PT, and TT were significantly prolonged in TXL groups compared with the model group (P<0.05 or P<0.01), and FIB levels were significantly decreased compared with the model group (P<0.01). CONCLUSIONS TXL had a protective effect on TAO mice, and the mechanism may involve inhibition of thrombosis and inflammatory responses. TXL may be a potential drug for the treatment of TAO.
Mice ; Male ; Animals ; Thromboangiitis Obliterans/chemically induced* ; Interleukin-6/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Thrombosis

OBJECTIVETo investigate the expression of nuclear factor kappa B (NF-kgr;B) and tumor necrosis factor &alpha; (TNF-&alpha;) in lung tissue of acute paraquat poisoned rats.

METHODS68 male Wistar rats were randomly divided into 2 groups: the control group (n = 8), the intoxication group (n = 60). On the 1st, the 3rd, the 7th, the 14th and the 28th day after intoxication, the expression of NF-κB p65 and TNF-&alpha; in lung tissue were detected by LSAB immunohistochemistry (IH) staining. Meanwhile, the level of malondialdehyde (MDA) in plasma, and lung homogenate, the content of malondialdehyde (HPY) in lung homogenate were detected.

RESULTSThe levels of MDA in plasma on the 1st, the 3rd, the 7th day and in lung homogenate on the 1st, the 3rd day of the intoxication group [in plasma: (10.15 ± 3.15), (6.97 ± 1.65) and (5.44 ± 0.66) nmol/ml; in lung homogenate: (10.20 ± 2.43), (10.71 ± 171) nmol/ml] were significantly higher than that of the control group [in plasma: (3.84 ± 1.04) nmol/ml, in lung homogenate: (7.66 ± 0.66) nmol/ml]. The content of HPY in lung homogenate on the 14th and the 28th day after intoxication [(19.98 ± 2.86), (26.06 ± 4.06) µg/0.1 g lung homogenate] were higher than that of the control group [(8.80 ± 1.26) µg/0.1 g lung homogenate] significantly. The expression of NF-κB p65 and TNF-&alpha; in lung tissue were both significantly increased on the first day and the 3rd day of the intoxication group compared with the control group and weakened obviously after the 7th day.

CONCLUSIONAcute paraquat poisoning can induce increased expression of both NF-κB p65 and TNF-&alpha; in lung tissue; the enhanced activity of NF-κB may take part in the process of pulmonary injury in PQ poisoning.

Animals ; Lung ; metabolism ; pathology ; Male ; Paraquat ; poisoning ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism