1.Effects and Mechanism of Xiaozheng Decoction on the Proliferation and Migration of Uterine Leiomyoma Cells in Rat
Yanli WU ; Kai FAN ; Shoufang KONG ; Xiufang ZHAO ; Jianghua OUYANG
China Pharmacy 2021;32(20):2450-2458
OBJECTIVE:To study the effects a nd mechanism of X iaozheng decoction on the proliferation and migration of uterine leiomyoma cells in rat. METHODS :Female SD rats were randomly divided into normal control group ,model group , chemical medicine positive control group (Mifepristone tablets ,2.25 mg/kg),TCM positive control group (Guizhi fuling capsules , 200 mg/kg),Xiaozheng decoction low-dose ,medium-dose and high-dose groups (1.4,2.8,5.6 g/kg),with 15 rats in each group. Except for normal control group ,other groups were given intramuscular injection of estrogen and progesterone to induce uterine leiomyomas model. On the second day after modeling ,rats in administration groups were given relevant medicine intragastrically ; normal control group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 months. After last administration ,the uterus was removed and its morphology was observed ;the uterine coefficient was calculated. Uterine leiomyoma cells or uterine smooth muscle cells were isolated and cultured. The proliferation rate and migration rate of cells were detected by MTT method and cell scratch test ;flow cytometry was used to detect the cell apoptosis rate ;mRNA expression of HMGB 1 were detected by RT-qPCR. The expression of phosphorylated protein kinase B (p-Akt),nuclear factor κB inhibitor α (IκBα)and phosphorylated transforming growth factor β-activated kinase 1(p-TAK1)were detected by ELISA ;the protein expression of HMGB 1,phospholipid 3 kinase(PI3K),p-Akt in cytoplasm and nuclear factor κB p65(NF-κB p65)in nucleus were detected by Western blot assay. RESULTS :Compared with normal control g roup,the myometrium of the model group was significantly thickened ,the number of uterine smooth muscle cells were significantly increased and the sizes were 83777930。E-mail:jianghua_ouyang@126.com different,the arrangement of muscle fibers in so me areas was disordered,the uterine coefficient ,and the relative expression of HMGB 1 mRNA and protein were increased significantly (P< 0.01). Compared with model group ,the thickening of uterine myometrium and other symptoms were improved to different extents in Xiaozheng decoction groups and positive control groups ;the uterine coefficient ,cell proliferation rate ,migration rate ,mRNA and protein expression of HMGB 1,the expression of p-Akt and IκBα,protein expression of PI 3K and p-Akt (except for Xiaozheng decoction low-dose group )in cytoplasm were all decreased significantly (P<0.05 or P<0.01);the cell apoptosis rate ,the expression of p-TAK 1,protein expression of NF-κB p65 in nucleus were all increased significantly (P<0.05 or P<0.01). The effects of Xiaozheng decoction showed a dose-dependent trend. CONCLUSIONS :Xiaozheng decoction can inhibit the proliferation and migration of uterine leiomyoma cells by down-regulating the expression of HMGB 1,PI3K and p-Akt ,up-regulating the expression of NF-κB p65,so as to promote cell apoptosis.