Metabolomic Study on the Preventive Effect of Patrinia scabiosaefolia Fisch on Multipathogen Induced Pelvic Inflammatory Disease in Rats

Abstract

Patrinia scabiosaefolia Fisch (PSF), a well-known traditional Chinese medicine (TCM), has been used as a "heat-clearing and detoxifying" agent. The present study was to illustrate the preventive effect of PSF on pelvic inflammatory disease (PID) in rats. The PID model was constructed by multipathogen infection of the upper genital tract with reference to the method previously reported. Urine metabolomic analysis was conducted with a GC-MS coupled with derivatization method. In this study, PID rats showed obvious infiltration of inflammatory cells and elevated expression of cytokines (IL-1β and IL-6) in upper genital tract, compared with control rats. Sixteen differentiating metabolites contributed to the alteration of metabolic profile in PID rats, including two amino acids, three fat acids, nine organic acids, and two types of sugars. The rats, infected by multipathogen and administered with PSF, showed decreased infiltration of inflammatory cells and lowered expression of cytokines in upper genital tract, compared with PID rats. Meanwhile, PSF intervened in the PID-associated alterations in TCA cycle, sugar metabolism, amino acid metabolism, and other uncertain metabolic pathways. These results indicate that PSF has preventive effect on multipathogen induced PID and holistic interventional effect on disease-associated metabolomic change.

Figure 1

Histological changes in uterus and fallopian tube after pathogen infections and administration of PSF extract. Representative micrographs of uterus and fallopian tube stained by H & E (×100) are showed: (a) uterus of control group; (b) uterus of PID group; (c) uterus of PSF group; (d) fallopian tube of control group; (e) fallopian tube of PID group; and (f) fallopian tube of PSF group. The positive staining of neutrophil is indicated as ▲, and the lymphocyte cell infiltration is indicated as ↑. Histological semiquantitative scores of inflammatory cells infiltration in uterus (g) and fallopian tube (h) are presented. Each bar represents the mean ± SD (^∗∗ P < 0.01, significantly different when compared with PID group; n = 3).

Figure 2

Representative base peak intensity chromatogram of the rat urine based on GC-MS.

Figure 3

PCA scores plot based on the metabolomic data obtained from urine in 3 groups (R2X = 0.713, Q2 = 0.519; n = 8). The red squares indicate control group, the green triangles indicate PSF group, and the black diamonds indicate PID group.

Figure 4

OPLS-DA scores plot based on the metabolomic data obtained from urine in control group and PID group (R2X = 0.605, R2Y = 0.996, Q2 = 0.978; n = 8). The red squares indicate control group, and the black diamonds indicate PID group.

Figure 5

Representative mass spectrum of differentiating metabolite (a) and glycine standard (b).

Figure 6

PID-associated differentiating metabolite changes after pathogen infections and administration of PSF extract. The value of the y-axis represents mean normalized peak areas of these metabolites. Each bar represents the mean ± SD (^∗ P < 0.05, ^∗∗ P < 0.01, significantly different from control group; ^# P < 0.05, ^## P < 0.01, significantly different from PID group; n = 8).