Figure 1.
Structure of triptolide.
Table 1.
ESI-MS MS/MS MRM and HPLC conditions for each sphingolipid.
Figure 2.
a) The splenic index of each group. The splenic index is expressed as the ratio of the weight of the spleen weight to the weight of the mouse. The four groups include: control, DTH model, DTH model+triptolide and control+triptolide, each of which contains eight samples. Statistical differences detected between the control & DTH model groups and the DTH model & DTH model+triptolide groups are indicated by the P-value between each group. b) Ear swelling in each group. Ear swelling was expressed as the difference between the weight of left and right ear patches obtained from 8 mm punches 30 h after challenge. The punches were obtained in a blinded manner. Data are expressed as the mean ± SD, and the values for each sample are the mean of eight separate samples.
Figure 3.
Sphingolipid composition of mice liver or spleen measured by triple quadruples MS/MS.
Four groups including: control, model, model+triptolide and control+triptolide, each of which contains 8 samples. Sphingolipids were isolated from liver or spleen homogenate corresponding to 1 mg protein. Bars are expressed as means ± SD, values for each tissue sample are the average of 8 samples separately (pmol/mg protein). Statistical difference from control or model group is indicated with an asterisk or a “&”, respectively. * or &: p<0.05 and ** or &&: p<0.01.
Figure 4.
Visualization of sphingolipid metabolites in the plasma, kidney, spleen and liver.
The figure depicts sphingolipid metabolites that participate in the early steps of sphingolipid biosynthesis. The pathway maps are overlaid by the fold changes in subclasses of sphingolipid metabolites between the control group and other groups by asterisks indicating the statistical significance on the right side (* = P<0.05, ** = P<0.01; n = 8). The content of each sphingolipid metabolite was determined as the mean of eight independent parallel samples.
Figure 5.
Score plots from supervised OPLS-DA.
Analysis showed distinct clustering between each group in the spleen, kidney, liver and plasma. R2Y(cum) and Q2(cum) determined by Simca P+12.0.1 are shown under each panel and indicate the stability and predictability of the model.
Table 2.
Sphingolipid biomarkers found in spleen, liver, plasma and kidney.
Figure 6.
Sphingolipid composition of mice kidney or plasma measured by triple quadruples MS/MS.
Four groups including: control, model, model+triptolide and control+triptolide, each of which contains 8 samples. Sphingolipids were isolated from kidney homogenate corresponding to 1 mg protein or from 0.1 mL plasma. Bars are expressed as means ± SD, values for each sample are the average of 8 samples separately (pmol/mg protein, 0.1 mL plasma). Statistical difference from control or model group is indicated with an asterisk or a “&”, respectively. * or &: p<0.05 and ** or &&: p<0.01.
Figure 7.
Sphingolipid biomarkers of DTH disease severity and response to triptolide treatment.
The levels of seven potential sphingolipid biomarkers in the plasma (A, B) and spleen (C-G) in control, DTH model, DTH model+triptolide and control+triptolide groups. The sphingolipids were identified based on data obtained by OPLS-DA with a significant correlation (p<0.05) to ear swelling and splenic index as indicators of DTH severity. Significant differences between the two groups were evaluated by Mann-Whitney U test with the P-value shown above each plot.
Table 3.
Correlation of sphingolipid biomarkers with ear swelling and splenic index.