ABSTRACT
Objective To investigate the effects of different doses of gingko flavonoid on the expression of nuclear factor-κBp65 (NF-κBp65) and tumor necrosis factor-α(TNF-α) in mice with non-alcoholic fatty liver disease(NAFLD) .Methods The mice mod-el of NAFLD was constructed by the high fat diet for 4 weeks .The mice were randomly divided into the control group ,high fat model group ,high ,middle and low doses of gingko flavonoid groups .High ,middle and low doses of gingko flavonoid groups were respectively gavaged with the different doses of gingko flavonoid .The normal control group and the high fat model group were ga-vaged with normal saline .After 12 weeks ,the mice were killed for taking liver tissue and detecting NF-κBp65 expressions ,and ser-um TNF-αand triacylglycerol(TG) levels were detected .Results Compared with the normal control group ,the levels of NF-κBp65 ,TNF-αand TG in the high fat model group were increased(P<0 .05) .Compared with the high fat model group ,the levels of NF-κBp65 ,TNF-αand TG in the high ,middle and low doses of gingko flavonoid groups were decreased (P<0 .05) ,moreover the higher the dose ,the more obvious the decrease .Conclusion Gingko flavonoid may play certain role in the treatment of NAFLD by reducing the generation of NF-κBp65 ,TNF-αand TG .
ABSTRACT
Objective To investigate the effect of hypothermia on the expression Toll-like receptor 2 (TLR2),myeloid differentiation factor 88(MyD88),nuclear factor-κBp65(NF-κBp65),plasminogen activator inhibitor-1(PAI-1)in the TLR2/MyD88 pathway in rats with acute lung injury(ALI)induced by lipopolysaccharide (LPS)inhalation. Methods Ninety male Sprague-Dawley(SD)rats were randomly divided into control group (n=18),hypothermia group(n=24),temperature controlled group(n=24),and temperature-uncontrolled group(n=24). The ALI model was reproduced by 0.5 mL/kg LPS intratracheal instillation,while only normal saline was instilled intratracheally for control group. Arterial blood was collected and physical cooling was started 1 hour after instillation. The body temperature was lowered to 32-34 ℃in hypothermia group and 36-37 ℃in temperature controlled group,and no intervention was used for temperature-uncontrolled group and control group. The arterial blood gas was determined in all the groups before and 1 hour after instillation of saline or LPS and 1,6, 12 hours after intervention. Rats were sacrificed respectively at 1,6 and 12 hours after temperature control therapy, the morphological changes in lung tissue were observed under light microscope. The protein expression of PAI-1 in bronchoalveolar lavage fluid(BALF)was determined by enzyme linked immunosorbent assay(ELISA). TLR2 mRNA and MyD88 mRNA transcriptional level were determined by reverse transcription-polymeras chain reaction (RT-PCR). NF-κBp65 protein level was determined by Western Blot. Results After instillation of LPS,the oxygenation index(PaO2/FiO2)of each group was decreased obviously,the damage of lung tissues was aggravating,the lung injury score was increased significantly,PAI-1 protein in BALF and the expressions of TLR2 mRNA,MyD88 mRNA, NF-κBp65 protein in lung tissues were increased obviously. Each index was improved by therapeutic Hypothermia, the effect of which was best in using a cooling period in the 1-6 hours,while might be benefit at 6-12 hours. Compared with temperature controlled group,PaO2/FiO2(mmHg,1 mmHg=0.133 kPa)at 1 hour and 6 hours of hypothermia group was improved(1 hour:402.49±38.61 vs. 324.36±28.93,6 hours:349.72±98.20 vs. 284.35±13.68, both P<0.01),the lung injury score at 1,6 and 12 hours were significantly decreased(1 hour:6.04±0.74 vs. 7.96±0.65,6 hours:9.09±0.80 vs. 13.13±1.02,12 hours:10.79±1.42 vs. 13.42±0.68,all P<0.01),the PAI-1 protein(ng/L)in BALF at 1,6 and 12 hours were significantly decreased(1 hour:121.36±4.62 vs. 197.74±9.42, 6 hours:230.53±10.76 vs. 294.06±16.60,12 hours:270.48±13.20 vs. 319.40±10.24,all P<0.01),TLR2 mRNA and MyD88 mRNA expressions(2-ΔΔCt)in the lung tissues at 1,6 and 12 hours were significantly decreased (TLR2 mRNA 1 hour:2.18±0.26 vs. 3.04±0.39,6 hours:4.09±0.29 vs. 4.90±0.35,12 hours:6.02±0.43 vs. 7.10±0.54;MyD88 mRNA 1 hour:2.25±0.41 vs. 3.04±0.30,6 hours:5.67±0.55 vs. 7.01±0.76,12 hours:7.14±0.60 vs. 8.87±0.54,all P<0.01),NF-κBp65 protein expression(A value)at 6 hours and 12 hours was significantly decreased(6 hours:0.31±0.08 vs. 0.53±0.12,12 hours:1.05±0.17 vs. 1.76±0.35,both P<0.01). There was no difference in each index between temperature controlled group and temperature-uncontrolled group. Conclusion Hypothermia can down-regulate the expression of TLR2 mRNA,MyD88 mRNA,NF-κBp65 protein and PAI-1 in the TLR2/MyD88 pathway to protect lung tissue of rats with ALI induced by LPS inhalation from injury.
ABSTRACT
Objective To explore the effects of matrine on apoptosis and nuclear factor(NF)-κBp65 activity of HL-60 cells induced by pirarubicin (THP).Methods Effects of the apoptosis:the HL-60 cells in blank control group were cultured 14 hours with RPMI 1640; groups of different concentration drugs:matrine:0.1 μmol/L,1.0 μmol/L,10.0 μmoL/L and THP:1.0 μmol/L,10.0 μmol/L,100.0 μmol/L.The apoptosis of HL-60 cells were tested by fluorescence double colour painting and flow cytometry (FCM).The NF-κBp65 activity of HL-60 cells were determined by FCM.Results The rates of apoptosis of HL-60 cells were (7.14 ± 2.95) ％,(12.34 ± 2.55) ％,(19.3 ±2.31)％ and (31.78 ±4.31)％,(47.25 ±5.27)％,(56.49 ±1.59)％ in matrine 0.1 μmol/L,1.0 μmol/L,10.0 μmol/L and THP 1.0 μmoL/L,10.0 μmol/L,100.0 μmol/L groups,compared to blank control group (6.46 ± 1.45) ％,there were significant difference (F =257.72,677.19 ; all P ＜ 0.001) ; (70.17 ± 5.68) ％ in matrine 0.1 μ mol/L +THP 1.0 μ moL/L group[vs the THP 1.0 μmoL/L group (31.78 ±4.31)％,(t =38.94,P＜0.001)].The rates of NF-κBp65 activity of HL-60 cells in matrine 0.1 μmol/L,1.0 μmol/L,10.0 μmol/L and THP 1.0 μmol/L,10.0 μmol/L,100.0 μmol/L groups were(8.34 ± 1.52)％,(7.11 ± 1.29)％,(4.78 ±0.31)％ and (16.21 ± 1.20) ％,(23.98 ± 3.21) ％,(32.44 ± 2.89) ％,with the control group (8.44 ± 2.20) ％,there were significant difference(F =65.35,P ＜ 0.001 ; F =674.11,P ＜ 0.001) ; (12.01 ± 2.27) ％ in matrine 0.1 μmol/L + THP 1.0 μmol/L group [with (16.21 ± 1.20) ％ of THP 1.0 μmol/L group(t =21.42,P ＜ 0.001)].Conclusions The apoptosis of HL-60 cell is induced by matrine and pirarubicin in a dose-dependent manner.NF-κBp65 activity of HL-60 cell is inhibited by matrine and increased by pirarubicin in a dose-dependent manner.Matrine can enhannce the effect of induction by pirarubicin of apoptosis of HL-60 cell,and decrease activity of NF-κBp65 by pirarubicin.