Effect Of Juantong Decoction On TLR4/NF-?B Signaling Protein In Eutopic Endometrial Cells Of Endometriosis

Objective: To study the effect of Juantong decoction on the proliferation and apoptosis of endometrial stromal cells(ESC)in endometriosis(EMs),observing the effect of Juanjianyin on expression of TLR4 / NF-?B signaling pathway in the expression of related factor protein to clarify the role of Juan pain drink on the treatment of endometriosis at the same time.Methods: Cultured the primary endometrial stromal cells in patients with endometriosis in vitro,take the ESC of the third generation to the 5th generation as the experimental study object: 1.Use Juantong decoction(high,medium and low dose),gestrinone,Chinese medicine Guizhi Fuling pill containing serum to deal with ESC.To observe the cell growth status,cell morphology and ultra structural changes by inverted microscope and transmission electron microscopy(TEM).Use the CCK-8 method to detect the inhibitory effect on the cells after 24 h,48 h and 72 h,and then draw the cell's growth curve;The effects of serum on the cell cycle and apoptosis were measured by flow cytometry(FCM).2.The expression of NF-?B,TNF-? and IL-1 protein in ESC which was intervened by PDTC was detected by Western blotting(WB).To detect the binding ability of NF-?B to DNA by gel electrophoresis mobility assay(EMSA).Results: 1.Cell proliferation inhibition rate comparison: The inhibitory rate of cell proliferation was significantly increased in each drug-containing serum group,and the difference was statistically significant(P <0.05),the inhibitory effect of Juantong decoction on cell proliferation was time-concentration-dependent,that's say the effect of the high dose group was the most significant,compared with western medicine group and Chinese medicine group,the difference was statistically significant(P <0.05).2.Cell cycle changes: the ESC of each drug-containing serum group were inhibited to varying degrees from the G1 phase to the S phase,compared with the blank group,the difference was statistically significant(P <0.05),Among them,the cell inhibition of Juantong decoction drug serum was concentration-dependent,that's say the effect of the high dose group was the most significant,compared with western medicine group and Chinese medicine group,the difference was statistically significant(P <0.05).3.Changes in apoptosis: Each drug-containing serum group have a significant role in promoting apoptosis of ESC,compared with the blank group,the difference was statistically significant(P <0.05),among them,the cell apoptosis of Juantong decoction drug serum was concentration-dependent,that's say the effect of the high dose group was the most significant,compared with western medicine group and Chinese medicine group,the difference was statistically significant(P <0.05).4.TLR4,NF-?B,IL-1 protein expression changes: Each drug-containing serum group has an inhibitory effect of the expression of TLR4,NF-?B and IL-1 protein in ESC,compared with the blank group,the difference was statistically significant(P <0.05),Among them,the inhibitory effect of Juantong decoction drug serum on cell protein was concentration-dependent,that's say the effect of the high dose group was the most significant,compared with western medicine group and Chinese medicine group,the difference was statistically significant(P <0.05).The high dose of Juantong decoction drug serum could decrease the expression of TLR4 and NF-?B protein but raising the expression of IL-1 protein after PDTC intervention,the difference was statistically significant(P <0.05).5.NF-?B and DNA binding capacity changes: Each drug-containing serum group could down-regulate the binding ability of NF-?B to DNA,the effect of the high dose group was the most significant,the binding capacity of NF-?B and DNA was significantly decreased after PDTC intervention.Conclusions: 1.Juantong decoction serum can change the ultra structure of ESC,inhibit cell proliferation,promote cell apoptosis,the inhibitory effect of high dose group was the most significant,compared with the blank group,the difference was statistically significant(P <0.05),2.Serum with Juantong decoction can reduce the expression protein of the ESC in TLR4,NF-?B,IL-1;the effect of high dose group was the most significant.And can reduce the binding capacity of ESC NF-?B and DNA;Inhibit the activity of NF-?B,Reduce the expression of NF-?B protein,Thereby inhibiting the expression of inflammatory factors,chemokines,adhesion molecules,apoptotic genes,Inhibition of ectopic endometrial invasion,adhesion and angiogenesis,and ultimately achieve the purpose of treatment of EMs.3.One of the mechanisms of Juantong decoction in treating EMs may be through the regulation of TLR4 / NF-?B signaling pathway.