Mechanism of Wenjingtang on Improving Endometriosis in Rats Based on Regulating HIF-1α Expression / 中国实验方剂学杂志

ABSTRACT

Objective:To observe the effect of Wenjingtang on the expression of hypoxia inducible factor-1α （HIF-1α） and ultrastructure of mitochondria in rats with endometriosis （EMs）， and to explore the mechanism of Wenjingtang in the treatment of EMs. Method:The EMs model rat was established by autogenous intimal transplantation， and the volume of ectopic lesions was measured by small animal ultrasound imaging system. According to the volume of ectopic lesions， rats successfully modeled were randomly divided into model group， low， middle ang high dose Wenjingtang group （4.85， 9.7， 19.4 g·kg-1） and gestrinone group （0.25 mg·kg-1）， 10 in each group， another 10 rats were recruited in a sham operation group. After 6 weeks of drug treatment， the volume of ectopic lesions was measured by ultrasound imaging system and caliper， the morphology of ectopic endometrium was observed by hematoxylin-eosin （HE） staining， the levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and transforming growth factor-β1 （TGF-β1） in peritoneal fluid were measured by enzyme linked immunosorbent assay （ELISA）， the mRNA and protein expression of HIF-1α in eutopic or ectopic endometrial tissues were detected by Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， and the ultrastructure of mitochondria in ectopic endometrium was observed by transmission electron microscope （TEM）. Result:Compared with sham operation group， ectopic lesions were found in model group， showing vesicular structure， and the typical endometrial tissue morphology was found in the lesions. The contents of IL-1β， TNF-α， TGF-β1 and the expression of HIF-1α mRNA and protein were significantly increased than those in the sham operation group （P<0.05，P<0.01）. TEM showed that the number of mitochondria in the cytoplasm of ectopic endometrium increased and the structure was intact. Compared with model group， the volume of ectopic focus was significantly decreased in all Wenjingtang groups （P<0.01）， and the results of ultrasonic examination were basically consistent with those of caliper measurement， HE staining showed that the columnar epithelial cells of ectopic endometrium were damaged or exfoliated and the interstitial cells were loose， the content of TNF-α in each dose group of Wenjingtang was significantly decreased， the content of IL-1β and TGF-β1 in the medium dose and high dose groups of Wenjingtang was significantly decreased （P<0.05，P<0.01）， the expression of HIF-1α mRNA and protein were significantly decreased in all Wenjingtang groups （P<0.05，P<0.01）. The mitochondria of ectopic endometrium were obviously swollen， the crest was broken or even disappeared， some of the mitochondria were vacuolar degeneration and the outer membrane was ruptured. Conclusion:Wenjingtang has a good therapeutic effect on experimental EMs in rats， and the mechanism is related to reducing the expression of HIF-1α， improving hypoxia in ectopic lesions and inducing mitochondrial damage in ectopic endometrium.