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Aim To investigate the effects of Kudino- side D on lipid accumulation induced by oxidized low density lipoprotein ( ox-LDL) and inflammation induced by lipopolysaccharide ( LPS ) in RAW264.7 cells.Methods Foam cells were established by incubating the RAW264.7 cells with ox-LDL.The concentration of lipid droplets in the cells was observed by oil red staining, and the level of total cholesterol (TC) in cells was measured by enzyme method.The gene and protein expressions of scavenger receptors CD36 and SR-A1, ATP binding cassette transporters A1 and Gl ( ABCA1 and ABCGI) were detected by RT-qPCR and Western blot, respectively.The expressions of inter- leukin-6 (IL-6), interleukin-1 (3 (IL-ip), monocyte chemoattractant protein-1 (MCP-1 ) and tumor necrosis factor-a (TNF-a) were detected by ELISA and RT-qPCR.The protein expressions of mTOR and p-mTOR were detected by Western blot.Results Compared with model group, the high dose of Kudinoside D decreased the content of TC and down-regulated the gene and protein expression of SR-A induced by ox-LDL.Meanwhile Kudinoside D also decreased the levels of IL-ip and MCP-1 and down-regulated the protein expression of p-mTOR induced by LPS.Conclusions Kudinoside D may reduce the intracellular TC content by down-regulating the gene and protein expression of SR-A1.Kudinoside D may play an anti-inflammatory role through mTOR pathway.
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Aim To investigate the interactions be-tween the neuroinflammation caused by lipopolysaccha-ride(LPS) and brain CYP2E1.Methods The human cholinergic neuroblastoma cell line IMR-32 was treated with LPS ( 0.1 mg · L-1 , 1.0 mg · L-1 ) , and the LDH and SOD activities were determined after 24 h in-cubation .In order to determine the roles of MAPK sig-naling pathway in the regulation of CYP 2E1 by LPS, the IMR-32 cells were treated with p38 pathway inhibi-tor SB203580 or ERK pathway inhibitor U 0126 for 45 min before the incubation with LPS .The human do-paminergic neuroblastoma cell line SH-SY5Y with CYP2 E1 over-expression was established . The LDH and SOD activities were determined in SH-SY5 Y cells over-expressed CYP2 E1 and control cells treated with LPS(0.1 mg· L-1 , 1.0 mg· L-1 ) for 24 h.Results The levels of LDH in IMR-32 cells treated with high-dose LPS were increased by 1.38-fold ( P <0.01 ) compared with the control group , and the levels of SOD reduced by 15.0%( P <0.01 ) .Compared with the control, CYP2E1 mRNA and protein levels in IMR-32 cells treated with high-dose LPS were increased by 1.25-fold(P<0.01) and 1.19-fold(P<0.05).The up-regulation of CYP2E1 by LPS could be attenuated by SB203580 and U0126 pretreatment.Compared with the control cells, the CYP2E1 over-expression in-creased LDH levels by 1.28-fold ( P<0.01 ) and de-creased SOD levels by 3.53-fold ( P<0.01 ) after the low-dose of LPS treatment .The CYP2E1 over-expres-sion increased LDH levels by 1.54-fold ( P <0.01 ) and decreased SOD levels by 2.17-fold( P<0.01) af-ter the high-dose of LPS treatment , compared with the control cells.Conclusions LPS can induce CYP2E1 mRNA and protein levels , and the p38 and ERK sig-naling pathway may be involved in the regulation .The elevated CYP2 E1 levels aggravate the damage to neuro-nal cells caused by LPS .Brain CYP2E1 may be an im-portant contributing factor to the pathological process of neuroinflammatory injury .
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Aim Toinvestigatetheeffectofbencyclo-quidium bromide(BCQB)on mucus MUC5AC expres-sion induced by lipopolysaccharide in cultured human nasalepithelialcells(HNECs).Methods Primary culture of human nasal epithelial cells (HNECs)was randomly divided into control group (C,with no treat-ment),LPS group (LPS,with LPS 1 mg · L-1 added in),BCQB low dose group(BCQBL,with LPS 1 mg· L-1 and BCQB 10 -8 mol·L-1 added in),BCQB mid-dle dose group(BCQBM,with LPS 1 mg·L-1 and BC-QB 10 -7 mol·L-1 added in),BCQB high dose group (BCQBH,with LPS 1 mg·L-1 and BCQB 10 -6 mol· L-1 added in)and ipratropium bromide group(IB,with LPS 1 mg·L-1 and IB 10 -6 mol·L-1 added in).Af-ter incubation at 37 ℃with 5% CO2 for 24 h,the ex-pression of MUC5 AC mRNA was detected with Real-time PCR and the expression of MUC5 AC protein in HNECs was detected with Western blot,while the ex-pression of MUC5 AC protein in supernatant was detec-tedwithELISAineachgroup.Results Ascompared with control group,the expression of MUC5 AC mRNA and protein increased significantly in LPS group (each P0. 05 ). Conclusion Bencycloquidiumbromidecansuppress MUC5 AC expression induced by LPS in cultured hu-man nasal epithelial cells,indicating that BCQB may be a new drug for nasal mucous hypersecretion diseases.
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Objective To investigate the effect of radix paeoniae rubra (RPR) on expressions of p38MAPK, NF-κB and iNOS in ltpopolysaccharide (LPS)-induced acute lung injury (ALI) and explore its protective mechanism. Methods Forty Wistar rats with ALI were divided randomly into five groups: saline control group (Group A) , LPS group (Group B), RPR for treatment group (Group C), RPR for prevention group (Group D) and SB203580 group (Group E). The effects of RPR on protein content and the ratio of neutrophiles in bronchoalveolar lavage fluid (BALF), contents of malondidehyde (MDA) and serum NO in lung tissue were observed. Arterial blood was collected for blood gas analysis. Expressions of p38MAPK, NF-κB and iNOS in rat lung tissues were detected. Results Compared with Group A, expressions of p38MAPK, NF-κB and iNOS were significantly increased (P <0.01 orP< 0.05) , the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly higher (P<0.01 or P<0.05) in Groups B, C, D and E. There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Groups B, C, D and E (P<0.01 or P<0.05). Compared with Group B, the expressions of p38MAPK, NF-κB and iNOS were significantly lower, and the protein content and the ratio of neutrophiles in BALF, contents of MDA and serum NO were significantly decreased, while the levels of arterial bicarbonate and partial pressure of oxygen were significantly higher in Groups C, D and E (P<0.05 or P<0.01). Compared with Group B, the lunginjury of Groups C, D and E was significantly alleviated, while there was no statistical difference among Groups C, D and E in the indices of lung injury. Conclusion Protective effect of RPR on LPS-induced ALI is closely related to the inhibited expressions of p38MAPK, NF-κB and iNOS.
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Objective To investigate the regulation of protein kinase C(PKC) to the expression of β-1,4-galactusyhransferase- Ⅰ ( β-1,4-GalT- Ⅰ ) and the influence on cytoskeleton and adherence ability of human umbilical vein endothelial cells(HUVECs) when stimulated by lipopolysaccharide (LPS). Methods Cultured HUVECs were pretreated by various PKC inhibitors or phorbol 12-myristate 13-acctate( PMA), an excitomotor of PKC respectively for 30 min, then stimulated by LPS for 4 h. β-1,4-GalT-Ⅰ expression were detected by RT-PCR and Western blot, expression of β-1,4-galactosylated carbohydrate chains and cytoskeleton were assayed by immumofluorescence, and adherence ability of HUVECs was observed by endothelialmonocyte cell adherence test. Results Up-regulated expression of β-1,4-GalT- Ⅰ and β-1,4-galactosylated carbohydrate chains in HUVECs stimulated by LPS were suppressed by PKC inhibitors and increased by PMA. F-actin and β-1,4-GalT- Ⅰ were partly co-localized in HUVECs. PKC inhibitor inhibited the effect of LPS on the distribution of F-actin and β-1,4-GalT- Ⅰ. Adherence ability of HUVECs enhanced by LPS was significantly suppressed by PKC inhibitor. Conclusion PKC signal transduction pathway may participate in regulating β-1,4-GalT-Ⅰ expression in endothelial cells stimulated by LPS. Furthermore, polytypes of PKC may participate in this regulating process; PKC might regulate cytoskeleton reorganization and adherence ability of EC through β-1,4-GalT-Ⅰ during inflammation.