Abstract
Purpose: To investigate the effect of Kang fu yan capsule (KFYC) on phenol mucilage-induced intrauterine adhesion (IUA) in a rat model, and the underlying mechanisms.
 Methods: An IUA model was established by injecting 0.06 mL of 25 % phenol mucilage into the uterus of female Sprague-Dawley rats. The IUA model rats (n=59) were randomly divided into 5 groups: IUA group, fuke qianjin tablet group (FKQJT, 0.22 mg/kg), and 3 KFYC groups given different doses of the drug i.e. 0.13, 0.39and 1.17 mg/kg. A group of 10 healthy female rats served as control. After 19 days treatment, blood samples were collected for determination of IL-2 and IL-10 by ELISA, while uterine tissues were subjected to histological examination using hematoxylin and eosin staining (H&E) and Masson staining. Expressions of Notch1, recombination signal binding protein-JK (RBP-JK), a disintegrin and metalloprotease (ADAM)-12, ADAM-15, matrix metalloprotein-9 (MMP-9), and inhibitor of NF-κB (IĸB) in uterine tissues were determined using RT-qPCR and western blot analysis.
 Results: Compared to IUA group, histological results showed reduced inflammatory cell infiltration in rat uterine tissue of KFYC group. Moreover, KFYC significantly reversed uterine fibrosis (p < 0.05). Serum concentrations of IL-2 significantly decreased in KFYC groups (p < 0.05 or p < 0.01), and there was significant increases the serum concentrations of IL-10 in KFYC groups (p < 0.05 or < 0.01), when compared to IUA group. The mRNA and protein expressions of Notch1, RBP-JK, ADAM-12, ADAM-15, MMP-9 were also significantly down-regulated (p < 0.05), while protein expression of IĸB was upregulated in KFYC group, when compared to IUA group.
 Conclusion: KFYC exerts an anti-IUA effect via amelioration of uterine inflammation and fibrosis, probably via a mechanism involving regulation of Notch1/ADAM pathway.
Highlights
Since 1900, intrauterine adhesions (IUAs) have been recognized as the cause of secondary amenorrhea
Results from real-time quantitative polymerase chain reaction (RT-qPCR) showed that the Notch1/a disintegrin and metalloprotease (ADAM) pathway was associated with IUA
In order to further investigate the exact role of Kang fu yan capsule (KFYC) in the IUA model, the protein expressions of Notch1, matrix metalloprotein-9 (MMP-9), IκB, ADAM-12, and ADAM-15 were determined by Western blot assay
Summary
Since 1900, intrauterine adhesions (IUAs) have been recognized as the cause of secondary amenorrhea. Kang fu yan capsule (KFYC) is a patent Chinese medicine which is used in the treatment of pelvic inflammation. The fuke qianjin tablet (FKQJT) is an effective patent Chinese medicine which has been used in the treatment of pelvic inflammation as a positive control. In line with the manufacturer’s instructions, standard or sample (100 μL) was added to each well and incubated for 2 h at 37 °C. The medium in each well was aspirated and the wells were washed 5 times, followed by addition of tetramethylbenzidine substrate (90 μL) and incubation for 20 min at 37 °C in the dark. The bands were semiquantitatively analyzed with Image J , and the results are expressed as ratios of Notch, IκB, MMP-9, ADAM-12, ADAM-15 to GAPDH densitometry readings. Values of p < 0.05 were considered statistically significant
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