Effects of Callicarpa nudiflora Granules on the Proliferation and Apoptosis of Uterine Fibroid Cells

Abstract

This research was aimed to discuss and understand the effects and mechanisms of action of Callicarpa nudiflora granules on proliferation and apoptosis of uterine leiomyoma (UL) cells. Firstly, normal uterine myometrium (UM) and UL tissues were collected, and the levels of p-Akt and Phosphatase and Tensin Homolog (PTEN) in UL tissues were detected using immunohistochemistry. Next, the UL cells were successfully obtained using enzymatic digestion, and their identification was performed using alpha-smooth muscle actin ( α -actin) immunocytochemistry. Specifically, the cells were grouped into four: a control group (CG), a low-dose group (LDG, 50 mg/L Callicarpa nudiflora solution), a medium-dose group (MDG, 100 mg/L Callicarpa nudiflora solution), and a high-dose group (HDG, 200 mg/L Callicarpa nudiflora solution). Moreover, the proliferation of UL cells was assessed using the thiazolyl blue (MTT) assay, while cell apoptosis was analyzed using flow cytometry (FCT). Real-time fluorescent quantitative PCR (fq-PCR) and Western blot assay (WBA) were utilized to determine the PAI-1, P38, TGF- β 1, E-cadherin, and Vimentin in UL cells. The results revealed that the positive rate (PR) of p-Akt in the UL tissues was much higher to that in normal UM tissues ( P < 0.001). More than 90% of UL cells were positive for α -actin. The viabilities of UL cells in the Callicarpa nudiflora treatment groups were greatly weakened to that of untreated cells ( P < 0.05). Viability of UL cells in the HDG group was the lowest, showing a great difference with P < 0.01 to the LDG group and that with P < 0.05 to the MDG group, while that between the MDG and LDG groups exhibited a great difference with P < 0.05. AR of UL cells in CG group was sharply lower to that in the Callicarpa nudiflora treatment groups, showing great differences with P < 0.05, P < 0.01, and P < 0.001, respectively. AR of UL cells in HDG group was higher to the LDG group ( P < 0.01) and MDG group ( P < 0.05), and that in LDG group was lower and exhibited a great difference with P < 0.05 to the MDG group. The HDG, LDG, and MDG groups exhibited greatly lower TGF- β 1, PAI-1, and P38 to the CG group ( P < 0.05). In the HDG group, the TGF- β 1, PAI-1, P38, and Vimentin levels were greatly lower and presented a great difference with P < 0.01 to those in the CG group and LDG group. Additionally, E-cadherin in UL cells was elevated in the LDG and MDG groups to CG group, showing P < 0.05 and P < 0.01, respectively. Such findings indicated that the Callicarpa nudiflora granules can suppress proliferation of UL cells and promote their apoptosis, which may be associated with the TGF- β 1/P38/PAI-1 singling pathway (SPW).