| Objective To summarize Shen Minhe's mainly therapeutic method of ovarian cancer.And we preliminary discussed the molecular mechanism of the compound as a representative of this therapeutic method on regulating drug resistance and metastasis of ovarian cancer in vitro.Methods ?We collected the Shen's clinical cases between the year of 2009 and 2012,and established the corresponding database in EXCELL.Then we used SPSS 20.0 to do the descriptive frequency analysis,correlation coefficient analysis,cluster analysis and chi-square test to study the rules and characteristics of his medication,from the angles of drug number,four natures of drugs,the five flavors(sour,bitter,sweet,pungent,and salty),channel tropism,function,drug corelationship,cluster and so on.?We summarized the correlation between Shen Minhe's mainly therapeutic method and the season in ovarian cancer.?We preliminary observed that the drug(Compound FuLing Granule)serum function had an effect on the drug resistance and metastasis of Skov3/DDP,through the experiment of cell proliferation,IC50,RI,Transwell and so on.? We observed the protein phosphorylation and protein expression of PI3k?Akt?GSK-3p?NK-?B,when the Skov3/DDP cultured with the drug(Compound FuLing Granule)serum in different concentration and time.Results ?The drug number of Shen's cases is 9-12 mostly.The drugs are based on warm(four natures of drugs),sweet(the five flavors),spleen and lung meridians,with vital complement functions mostly.We extracted and purified 18 couplet medicines.?We used two classification fast clustering method of SPSS and gained 7 drugs clustering.We summarized as follows:category 1 treatment is method of promoting blood circulation and removing blood-stasis based on Guizhi Fuling Wan;category 2 treatment is method of nourishing and warming kidney-yang based on Guifu Dihuang Wan;category 3 treatment is method of soothing liver to regulate qi based on Xiao Chaihu Tang;category 4 treatment is method of replenishing qi to invigorate the spleen based on Yigong San;category 5 treatment is method of detoxifing and warming kidney yang;category 6 treatment is method of dissipating carbuncle and eliminating phlegm and stagnation;category 7 treatment is method of dissipating dampness and helping digestion.From comprehensive results of constituent ratio and frequency,we found the top three treatments is method of promoting blood circulation and removing blood-stasis,replenishing qi to invigorate the spleen and warming and invigorating kidney yang.?We classified the drugs of cases in to 9 categories according to traditional Chinese pharmacology,such as tonifying qi,tonifying blood,tonifying yin,tonifying yang,regulating qi,promoting blood circulation,warming interior,clearing heat and dissipating phelm and dampness.From comprehensive results of constituent ratio and frequency,tonifying qi is given priority in deficiency syndrome elements and promoting blood circulation is given priority in excess syndrome elements.? Combined with clinic situation,for treatment summarization,7 treatment clustering according to clustering method of SPSS is better than 9 function categories according to traditional Chinese pharmacology.?From solar terms of Vernal Equinox,Ching Ming,Summer Solstice,Autumnal Equinox and winter solstice,three treatment(promoting blood circulation,replenishing qi to invigorate the spleen,dissipating dampness and helping digestion)has significant differences(P<0.001)between different solar terms.The treatment of promoting blood circulation is top 1.Treatment in Vernal Equinox,Ching Ming,and winter solstice is continued with replenishing qi to invigorate the spleen,warming and invigorating kidney yang.Treatment in Summer Solstice is continued with dissipating dampness and helping digestion,warming kidney yang.Treatment in Autumnal Equinox is continued with replenishing qi to invigorate the spleen,soothing liver to regulate qi.Because the main treatment of ovarian cancer in promoting blood circulation,so the patients should pay attention to doctors' office visiting in Ching Ming and Winter solstice.?Compound FuLing Granule at the concentration of 12.5mg/ml decreased RI(resistant index)of Skov3/DDP from 2.97±0.56 to 1.58±0.31,showing the effect of drug resistant inhibition.The Compound showed a significant dose-effect and time-effect correlation on suppressing the proliferation of Skov3/DDP at the concentration of 12.5mg/ml?62.5mg/ml,through different IC50 of 43.49mg/ml?29.27mg/ml?14.76mg/ml?13.84mg/ml in different time point of 24h?48h?72h?96h.The Compound decreased the cell crosswise movement and the number of penetrating cell membrance significantly.?The Compound at the concentration of 6.25mg/ml?12.5mg/ml had the ability of reversing drug-resistance and penetrating cell membrane,which may be related to PI3k-Akt-GSK-3? cell signaling.Conclusion ?Shen's main treatment methods of ovarian cancer are promoting blood circulation,tonifying qi and warming yang.The patients should pay attention to doctors' office visiting in Ching Ming and Winter solstice.In addition the treatment methods of promoting blood circulation,tonifying qi and dissipating phelm and dampness should be applied differently in different solar terms.?The in vitro studies show that,the drug resistant and metastasis inhibition effect of Compound FuLing Granule,as the representative therapy of promoting blood circulation,tonifying qi and warming yang,may be related to PI3k-Akt-GSK-3? cell signaling. |
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