Anti-uterine Tumor Effect And Characterization Of Volatile Oil Constituents In Prunella Vulgaris L

Objective: To find the best extraction process of supercritical fluid(CO2)extraction of Prunella vulgaris L,and to characterize its chemical composition,evaluate the pharmacodynamics and mechanism of its anti-uterine myoma,and provide basis for the clinical treatment of uterine myoma.Methods: 1)The volatile oil of Prunella vulgaris L was extracted by the supercritical fluid(CO2)extraction unit,the process was optimized.2)The chemical composition of the volatile oil of Prunella vulgaris L was analyzed by GC-MS.3)The MTT method was used to screen and evaluate the toxicity of the volatile oil of Prunella vulgaris L.4)Using the double fluorescent cell counting instrument to detect the volatile oil of Prunella vulgaris L to promote the apoptosis of uterine myoma cells and to block the growth cycle of the uterine myoma cells.5)the anti-uterine myoma activity of the volatile oil of Prunella vulgaris L was investigated by using the SD rat model of uterine myoma.6)The TUNEL and PCNA staining methods were used to detect the apoptosis of the volatile oil of Prunella vulgaris L and the inhibition of the proliferation of uterine myoma.7)The level of estrogen and progesterone in the serum of SD rats was detected by Elisa kit.8)The expression of estrogen,progesterone receptor,Survivin,Caspase-3,Bax and Bcl-2 in the uterus of SD rats was detected by immunohistochemical method.Result: 1)Volatile oil is one of the main components of Prunella vulgaris L.Supercritical fluid(CO2)extraction is an effective method for extracting volatile oil from Prunella vulgaris L.The optimum extraction process is 120 min,extraction pressure of 25 Mpa,and extraction temperature of 30.2)There are 16 kinds of volatile oil in Prunella vulgaris L.The main components are squalene(28.03%),linoleic acid(9.96%),linolenic acid(9.95%),stearic acid(6.26%)and oleic acid(5.51%).3)The results of in vitro experiment showed that the volatile oil of Prunella vulgaris L has significant anti human uterine myoma cell activity and no drug toxicity,and its mechanism is to promote the apoptosis of human uterine myoma cells,inhibit the uterine myoma cells in the G0-G1 stage,make the cells unable to enter the G2 stage for DNA replication,and inhibit the proliferation of uterine myoma cells.4)The experimental results showed that the volatile oil of Prunella vulgaris L had significant anti uterine leiomyoma activity.Its main mechanism was that the volatile oil of Prunella vulgaris L obviously inhibited the expression of estrogen and progesterone receptor in the uterus of SD rats,thus inhibiting the combination of estrogen,progesterone and receptor to inhibit the proliferation of uterine fibroids,and the volatile oil of Prunella vulgaris L.It can also promote the apoptosis of uterine myoma cells in SD rats,inhibit the proliferation of uterine myoma cells,reduce the overexpressed Survivin and Bcl-2 protein levels,inhibit the expression of Caspase-3 and Bax protein levels,and thus promote the apoptosis of uterine myoma cells.Conclusion: The volatile oil of Prunella vulgaris L has obvious anti uterine fibroid activity and is safe and non-toxic.It can be used as an ideal anti uterine fibroid candidate drug.