| Uterine leiomyomas are the most common benign tumors in women of reproductive age ,clinically diagnosed in 20 ~ 30% of women, however have been reported to occur at an incidence as high as 77%in women with or without clinical history of myometriums uteri [1].Although uterine leiomyomas are benign tumors,sympotommatic tumors are offen associated with infertility, spontanous abortion ,pelvic pain and menorrhagia.The homorne and operation treatment may also give rise to some significant side-effects such as menopausal syndrome,virilism ,premature-ovarian-failure,et al ,What's more ,cause the psychological disorders of the patients. Studies have shown that the traditional Chinese medicine of HUO XUE HUA YU could treat the uterine leiomyomas with seldom side-effects, My study aims at the mechanism of XIAO ZHENG WAN (XZW, abbreviated) treating leiomyomas at cell and molecular level and investigate the serum containing XZW effect on proliferation and early apoptosis in human uterine leiomyomas cells on basis of serum pharmacology. Objective:To investigate the serum containing-XZW effect on proliferation and apoptosis in human uterine leiomyomas cells. Design:The Wistar rats are fed on XZW decoctions ,with the dosage as ten times as human normal. Then stripple blood from the rats and contracts the serum containing-XZW, in terms of serum pharmacology principle .Use primary cultured human uterine leiomyomas cells as experimental cells in vitro. Devide cells into seven groups, add the different concetration rats serum containing -XZW(5% ?10%. 20%) as culture medium to the cells .compared to the complete medium(20%FBS) and the matched drug-free rats serum medium. After 24 hours,detect OD index and apoptosis of the all cells with MTT , Annexin V-FITC/PI respectively, while the nistological examination is although done by microscopy. Results:1 Serum containing-XZW effect on proliferation in human uterine leiomyomas cells Histological examination:The cells' growths are different, they shrink ,detacth from the cultured wells in the Serum containing-XZW groups.The other groups cells hold then- nomarl shuttle-shape, attaching on the wells. MTT assay:(1)The inhibition rates of leiomyomas cells treated by the Serum containing-XZW are significantly higher than that treated by the complete medium and drug-free serum medium(p<0.05). Among the serum containing-XZW groups;the inhibition rate is statistically increased for 20% serum group compared to the other two groups (p<0.05). Serum containing-XZW could arrest leiomyomas cells proliferation.(2) The concentrations of the serum containing-XZW are statistically closelylinked to the inhibition rate ( Regression equation : Y=0.5389+0.1 08901nx, r = 0.9360,p<0.01 ) ,the relations between the two aspects is in line with the curve (Y=0.5389+0.108901nx). In some concentration area, the higher the concentrations are ,the higher the inhibition rate is.(3) With the regression analysis ,r2=0.8760, the changes of cells proliferation arrested majorly attributed to the changes of serum containing-XZW, making up for 87.60%.2 Serum containing-XZW responsiveness of cells in human uterine leiomyomas early apoptosis Histological examination:Serum containing-XZW groups have more quincunx apoptosis cells with cells compacted, nucleolus shattered, fluorescent-colored fragments irregulated called apoptosis body, apoptosis bodies are hardly seen hi complete medium group . Annexin V-FITC/PI assay :(1) the Serum containing-XZW groups are significantly higher than the other two groups (p<0.05). The proportion is statistically increased for 20% serum group compared to the other two groups (p<0.05),making up for 26.88%.(2) The proportions of the cells in the cycle of GI and S+G2 in Serum containing-XZW is statistically higher and lower than that of hi complete medium and drug-depleted serum medium groups,respectively (p<0.05). The proportions of is statistically increased and decreased respectively for 20% serum group Serum containing-XZW group compared to 5% ,but n... |
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