Huayu Sanjie Enema Liquid Relieves Pain in Endometriosis Model Rats by Inhibiting Inflammation, Peripheral Sensitization, and Pelvic Adhesion

Abstract

The objective of this study is to observe the effect of relieving pain of Huayu Sanjie enema liquid (HYSJ-EL) on endometriosis model rats and to explore its mechanism of action. Of 24 female Sprague Dawley rats, six were randomly selected as the sham operation group (normal control group). The remaining rats were used to establish rat models of endometriosis through autologous endometrial transplantation combined with estrogen injection. Successfully modeled rats were randomly divided into the model, indomethacin (Western medicine group), and HYSJ-EL (Chinese herbs group) treatment groups. The thermal pain threshold of rats was measured, and hematoxylin and eosin staining was used to observe pathological changes after sampling. Serum levels of prostaglandin E2 (PGE2), interleukin-6 (IL-6), macrophage inflammatory protein-2 (MIP-2), plasminogen activator inhibitor-1 (PAI-1), and transforming growth factor-β (TGF-β) were measured using an enzyme-linked immunosorbent assay (ELISA). Furthermore, the protein and mRNA expression levels of transient receptor potential vanilloid-1 (TRPV1) and tumor necrosis factor-α (TNF-α) in the endometrium and endometriotic lesions were measured using Western blotting and quantitative real-time PCR assays, respectively. Compared to the model group, the heat pain threshold of rats in the HYSJ-EL group was significantly increased (P < 0.01), and the serum levels of PGE2, IL-6, MIP-2, PAI-1, and TGF-β were significantly decreased (P < 0.01), as well as the expression of TRPV1 and TNF-α protein and mRNA in the tissue of the ectopic lesion was significantly decreased (P < 0.05). These results indicate that the Huayu Sanjie enema liquid exerts analgesic effects on endometriosis by inhibiting inflammation, peripheral nerve sensitization, and pelvic adhesion.

Figure 1

Flowchart of the animal experiments.

Figure 2

The thermal pain threshold at 4 and 8 weeks.  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.  ^∧∧P < 0.01, compared to the indomethacin group.

Figure 3

The morphology of the endometrial tissue of each group. Black arrows show epithelial cells; blue arrows show glands.

Figure 4

The size of the lesions in each group. In the bar charts,  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.

Figure 5

The lesion fibrosis in each group. In the bar charts,  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.

Figure 6

Serum levels of PGE2, IL-6 and MIP-2. In the bar charts,  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.  ^∧∧P < 0.0, compared to the indomethacin group.

Figure 7

The serum levels of TGF-β and PAI-1. In the bar charts,  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.  ^∧∧P < 0.0, compared to the indomethacin group.

Figure 8

TRPV1 and TNF-α protein and mRNA expression in the endometriotic tissues. In the bar charts,  ^∗P < 0.05, compared to the normal control group.  ^∗∗P < 0.01, compared to the normal control group.  ^##P < 0.01, compared to the model group.  ^∧∧P < 0.01, compared to the indomethacin group.