Background:Polycystic ovary syndrome(PCOS)is one of the most common reproductive endocrine metabolic diseases.Combined use of metformin and diane-35 has better curative effect in regulating serum hormone level(LH,FSH,...Background:Polycystic ovary syndrome(PCOS)is one of the most common reproductive endocrine metabolic diseases.Combined use of metformin and diane-35 has better curative effect in regulating serum hormone level(LH,FSH,T and E2)than using metformin alone.Traditional Chinese medicine(TCM)can also be used to treat PCOS.According to some studies,the combined use of metformin and diane-35 and TCM have achieved better curative effect than combining metformin and diane-35 in the treatment of patients with PCOS.Methods:Computerized searches of the science,Medline,VIP,Wan Fang and China HowNet(CNKI)databases were conducted to identify eligible randomized controlled trials(RCTs)from the data obtained up to March 1,2022.The Cochrane Collaboration risk of bias tool was used to assess the risk of bias in individual RCTs,and R software(version 4.0.3)was used for data statistical analysis.Results:Nine RCTs involving 1035 patients were included.Comparing to D+M,significant reduce of LH(mean difference[MD]:-1.93,95%confidence interval[CI]:-3.44,-0.42;Unit:U/L P<0.01;I2=89%)、T(MD:-1.44,95%CI-2.59,-0.30;Unit:nmol/L P<0.01;I2=98%)and significant increase of E2(MD:31.43,95%CI 24.54,38.33;Unit:pmol/L P<0.01;I2=96%)were shown in TCM+D+M.Comparing to D+M,TCM+D+M group has higher ovulation rate(RR 1.1495%CI 1.07,1.22;P=0.42;I2=0%)and higher pregnancy rate(RR 1.2995%CI 1.15,1.44;P=0.37;I2=7%).There is no significant difference between the two therapies in FSH changes(MD:-1.00,95%CI-2.27,0.28;Unit:U/L P<0.01;I2=95%).Subgroup analysis showed that compared with the Guizhi Fuling capsule group,the Kuntai capsule group had more FSH reduction and E2 increase more.In other outcome indicators,the two subgroup did not show significant differences.Conclusion:Kuntai Capsule+Diane-35+Metformin is better than Guizhi Fuling Capsule in reducing FSH,and it is also better in increasing E2.There was no significant difference between the two in LH and T hormones.There was no significant difference between Kuntai Capsules+Diane-35+Metformin and Guizhi Fuling Capsules+Diane-35+Metformin.As for the effect in lessen insulin resistance,Kuntai Capsule+Diane-35+Metformin was significantly better than Guizhi Fuling Capsules+Diane-35+Metformin.展开更多
Guizhi Fuling capsule(GFC), a traditional Chinese medicine(TCM) with effects of promoting blood circulation and dissipating blood stasis, has been widely used in the clinic. Because of the complex matrix and various c...Guizhi Fuling capsule(GFC), a traditional Chinese medicine(TCM) with effects of promoting blood circulation and dissipating blood stasis, has been widely used in the clinic. Because of the complex matrix and various chemical structure types, quality control of GFC remains great challenge. In the present study, an ultra performance liquid chromatography hybrid triple-quadrupole mass spectrometry(UPLC-QQQ MS) method with ultrafast positive/negative ionization switching was developed for simultaneous determination of 18 bioactive components in GFC, including methyl gallate, ethyl gallate, oxypaeoniflorin, benzoic acid, albiflorin, paeonolide, paeoniflorin, 1, 2, 3, 4, 6-pentagalloylglucose, mudanpioside C, benzoyloxypaeoniflorin, benzoylpaeoniflorin, pachymic acid, amygdalin, cinnamaldehyde, paeonol, cinnamic acid, 4-hydroxybenzoic acid, and gallic acid. Separation was performed on an Agilent Zorbax Extend–C18 column(2.1 mm × 50 mm, 1.8 μm), using a gradient elution with acetonitrile and water containing 0.1% formic acid. Cholic acid was selected as the internal standard. This newly developed method was fully validated for linearity, precision, accuracy, and stability, and then applied to quality assessment of GFC. Finally, the batch-to-batch reproducibility of GFC samples was evaluated by the cosine ration and Euclidean distance method, which showed high quality consistency. The results demonstrated that the developed method provided a reasonable and powerful manner for quality control of GFC.展开更多
OBJECTIVE To evaluate the effect of Guizhi Fuling Capsule active pharmaceutical ingredient(API)and its fractions on human breast cancer cells proliferation by high-throughput screening assay.METHODS The crude fraction...OBJECTIVE To evaluate the effect of Guizhi Fuling Capsule active pharmaceutical ingredient(API)and its fractions on human breast cancer cells proliferation by high-throughput screening assay.METHODS The crude fractions were obtained from the extraction and elution of the API of Guizhi Fuling Capsule,and 929 standard fractions were obtained by the optimal separation conditions.Sulforhodamine B(SRB)method was used to evaluate the effects of the Guizhi Fuling capsule API and929 kinds of fractions on the proliferation of human breast cancer cells MCF-7 and MDA-MB-231.RESULTS The Guizhi Fuling capsule API had a strong ability to inhibit the proliferation of MCF-7 cells at high concentration and the ability to inhibit MDA-MB-231 cells' proliferate at low concentration following 72 h treatment;some samples of 929 fractions(5μg·mL^(-1))was found to have a breast cancer cell growth inhibition rate above 50%,without toxicity on HUVECs proliferation.CONCLUSION The API of Guizhi Fuling capsule had significant cytotoxicity effects on these two human breast cancer cells,with significant concentration-and time-dependent manner.展开更多
Objective To observe the effects of Guizhi Fuling capsule(GZFLC)on RPMI 8226 cells and explore the mechanisms.Methods Cell Counting Kit-8(CCK-8)assays and flow cytometry were used to detect the viability and apoptosis...Objective To observe the effects of Guizhi Fuling capsule(GZFLC)on RPMI 8226 cells and explore the mechanisms.Methods Cell Counting Kit-8(CCK-8)assays and flow cytometry were used to detect the viability and apoptosis levels of RPMI 8226 cells.The effects on mitochondria were examined by ROS and JC-1 assays.Western blotting was used to detect the expression of B cell lymphoma-2(Bcl-2),Bax,cleaved caspase-3,and Apoptotic protease-activating factor 1(Apaf-1).Results GZFLC drug serum decreased the viability and increased the apoptosis of RPMI 8226 cells.In addition,this drug increased ROS levels and decreased the mitochondrial membrane potential(MMP).Western blotting showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups,whereas the expression levels of cleaved caspase-3 and Apaf-1 were increased.Conclusion GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway.展开更多
Objective:To compare the clinical efficacy of Gongliuqing Capsule(宫瘤清胶囊)combined with mifepristone,Guizhi Fuling Capsule(桂枝茯苓胶囊)combined with mifepristone,and single mifepristone in the treatment of uterine...Objective:To compare the clinical efficacy of Gongliuqing Capsule(宫瘤清胶囊)combined with mifepristone,Guizhi Fuling Capsule(桂枝茯苓胶囊)combined with mifepristone,and single mifepristone in the treatment of uterine fibroids.Methods:Retrospective analysis was performed on 307 patients with uterine fibroids receiving drug treatment in our hospital.All 307 patients has been divided into three groups according to the different drug delivery scheme,GLM group of 125 patients taking Gongliuqing Capsule and mifepristone,GFM group of 113 patients taking Guizhi Fuling Capsule(桂枝茯苓胶囊)and mifepristone,SM group of 69 patients only taking mifepristone.Comparison of the cure rate and effective rate,volume of menstruation,menstrual period and uterine fibroids,level of estrogen and the incidence of adverse reaction of the patients of the 3 groups.Results:After 90days of treatment,the efficiency of GLM and GFM group were higher than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P>0.05).The cure rate of GLM group was higher than that of the other two groups,and the difference was statistically significant(P<0.05).The menstrual volume and period length,uterine fibroids volume of GLM and GFM group was smaller than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P<0.05).The P and E2 level of GLM and GFM group was lower than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P>0.05).There was no significant difference in the incidence of other sex hormone level and adverse reactions(P>0.05).Conclusions:Compared with the SM to treat uterine fibroids,the integrative medicine therapy were better.Among them,Gongliuqing Capsule(宫瘤清胶囊)combined with mifepristone has the highest cure rate.展开更多
Objective: To observe the effects of Guizhi Fuling Capsule(GZFLC) on myeloma cells and explore the mechanisms. Methods: MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell exp...Objective: To observe the effects of Guizhi Fuling Capsule(GZFLC) on myeloma cells and explore the mechanisms. Methods: MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell experiments were divided into negative(10%, 20% and 40%) groups, GZFLC(10%, 20%, and 40%)groups and a control group. Cell counting kit-8(CCK-8) assays and flow cytometry were used to detect the viability and apoptosis levels of myeloma cells. The effects on mitochondria were examined by reactive oxygen specie(ROS) and tetrechloro-tetraethylbenzimidazol carbocyanine iodide(JC-1) assays. Western blot was used to detect the expression of B cell lymphoma-2(Bcl-2), Bcl-2-associated X(Bax), cleaved caspase-3,-9, cytochrome C(Cytc)and apoptotic protease-activating factor 1(Apaf-1). RPMI 8226 cells(2×107) were subcutaneously inoculated into 48 nude mice to study the in vivo antitumor effects of GZFLC. The mice were randomly divided into four groups using a completely randomized design, the high-, medium-, or low-dose GZFLC(840, 420, or 210 mg/kg per day,respectively) or an equal volume of distilled water, administered daily for 15 days. The tumor volume changes in and survival times of the mice in the GZFLC-administered groups and a control group were observed. Cytc and Apaf-1 expression levels were detected by immunohistochemistry. Results: GZFLC drug serum decreased the viability and increased the apoptosis of myeloam cells(P<0.05). In addition, this drug increased the ROS levels and decreased the mitochondrial membrane potential(P<0.01). Western blot showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups, whereas the expression levels of cleaved caspase-3,-9,Cytc and Apaf-1 were increased(all P<0.01). Over time, the myeloma tumor volumes of the mice in the GZFLCadministered groups decreased, and survival time of the mice in the GZFLC-administered groups were longer than that of the mice in the control group. Immunohistochemical analysis of tumor tissues from the mice in the GZFLCadministered groups revealed that the Cytc and Apaf-1 expression levels were increased(P<0.05). Conclusion:GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway and significantly reduced the tumor volumes in mice with myeloma, which prolonged the survival times of the mice.展开更多
Purpose:The well-known traditional Chinese formula Guizhi Fuling capsule(GFC)has been reported to reverse ovarian cancer drug resistance.Extrachromosomal DNA(ecDNA)plays an important role in tumour metastasis and resi...Purpose:The well-known traditional Chinese formula Guizhi Fuling capsule(GFC)has been reported to reverse ovarian cancer drug resistance.Extrachromosomal DNA(ecDNA)plays an important role in tumour metastasis and resistance.The purpose of this study was to investigate the potential mechanisms by which GFC blocks tumour metastasis and reverses drug resistance by targeting ecDNA.Methods:CNKI and PubMed were used to obtain pharmacokinetic research data on GFC in rats,and the bioactive ingredients detected in rat serum or plasma were collected.Network databases were used to screen the abnormally expressed genes in ecDNA,tumour metastasis genes,resistance genes,and the active ingredient targets of GFC.The KOBAS3.0 database was used to enrich the KEGG pathways and GO functions;the STRING platform was used to construct the core protein interaction network;and the molecular docking online tool SwissDock was used to analyse the binding activity of the core targets and the active ingredients.RT-qPCR,Western blotting and laser confocal microscopy were used to verify the efect of the sera containing GFC on ecDNA,mRNA and protein expression of key targets.Results:Twenty-three bioactive ingredients of GFC were retrieved from PubMed and CNKI.Nine shared targets were simultaneously involved in abnormal genes in ecDNA,tumour metastasis and resistance and the active ingredient targets of GFC.GO functional analysis indicated that the cotargets involved cell proliferation,apoptotic regulation,nuclear functions,etc.The potential pathways involved in the reversal of tumour metastasis and drug resistance of GFC were the PI3K-Akt signalling,cancer,and platinum drug resistance pathways.Three shared proteins targeting ecDNA(AKT1,EGFR and MYC)stand out from the top 20 PPI targets,and all of the bioactive ingredients of GFC have strong binding afnity to the three proteins.The active ingredients can reduce the expression of MYC,EGFR and AKT1 mRNA and protein and the amount of ecDNA in drug-resistant OC cells.Conclusions:GFC targeting ecDNA to reverse tumour metastasis and drug resistance has the characteristics of multiple ingredients,multiple targets,and multiple pathways,which provides a new perspective for the development of new drugs targeting ecDNA to beneft tumour treatment.展开更多
文摘Background:Polycystic ovary syndrome(PCOS)is one of the most common reproductive endocrine metabolic diseases.Combined use of metformin and diane-35 has better curative effect in regulating serum hormone level(LH,FSH,T and E2)than using metformin alone.Traditional Chinese medicine(TCM)can also be used to treat PCOS.According to some studies,the combined use of metformin and diane-35 and TCM have achieved better curative effect than combining metformin and diane-35 in the treatment of patients with PCOS.Methods:Computerized searches of the science,Medline,VIP,Wan Fang and China HowNet(CNKI)databases were conducted to identify eligible randomized controlled trials(RCTs)from the data obtained up to March 1,2022.The Cochrane Collaboration risk of bias tool was used to assess the risk of bias in individual RCTs,and R software(version 4.0.3)was used for data statistical analysis.Results:Nine RCTs involving 1035 patients were included.Comparing to D+M,significant reduce of LH(mean difference[MD]:-1.93,95%confidence interval[CI]:-3.44,-0.42;Unit:U/L P<0.01;I2=89%)、T(MD:-1.44,95%CI-2.59,-0.30;Unit:nmol/L P<0.01;I2=98%)and significant increase of E2(MD:31.43,95%CI 24.54,38.33;Unit:pmol/L P<0.01;I2=96%)were shown in TCM+D+M.Comparing to D+M,TCM+D+M group has higher ovulation rate(RR 1.1495%CI 1.07,1.22;P=0.42;I2=0%)and higher pregnancy rate(RR 1.2995%CI 1.15,1.44;P=0.37;I2=7%).There is no significant difference between the two therapies in FSH changes(MD:-1.00,95%CI-2.27,0.28;Unit:U/L P<0.01;I2=95%).Subgroup analysis showed that compared with the Guizhi Fuling capsule group,the Kuntai capsule group had more FSH reduction and E2 increase more.In other outcome indicators,the two subgroup did not show significant differences.Conclusion:Kuntai Capsule+Diane-35+Metformin is better than Guizhi Fuling Capsule in reducing FSH,and it is also better in increasing E2.There was no significant difference between the two in LH and T hormones.There was no significant difference between Kuntai Capsules+Diane-35+Metformin and Guizhi Fuling Capsules+Diane-35+Metformin.As for the effect in lessen insulin resistance,Kuntai Capsule+Diane-35+Metformin was significantly better than Guizhi Fuling Capsules+Diane-35+Metformin.
基金supported by the Specialized Research Fund for the Doctoral Program of Higher Education(No.20130096140001)the 111 Project(No.B16046)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘Guizhi Fuling capsule(GFC), a traditional Chinese medicine(TCM) with effects of promoting blood circulation and dissipating blood stasis, has been widely used in the clinic. Because of the complex matrix and various chemical structure types, quality control of GFC remains great challenge. In the present study, an ultra performance liquid chromatography hybrid triple-quadrupole mass spectrometry(UPLC-QQQ MS) method with ultrafast positive/negative ionization switching was developed for simultaneous determination of 18 bioactive components in GFC, including methyl gallate, ethyl gallate, oxypaeoniflorin, benzoic acid, albiflorin, paeonolide, paeoniflorin, 1, 2, 3, 4, 6-pentagalloylglucose, mudanpioside C, benzoyloxypaeoniflorin, benzoylpaeoniflorin, pachymic acid, amygdalin, cinnamaldehyde, paeonol, cinnamic acid, 4-hydroxybenzoic acid, and gallic acid. Separation was performed on an Agilent Zorbax Extend–C18 column(2.1 mm × 50 mm, 1.8 μm), using a gradient elution with acetonitrile and water containing 0.1% formic acid. Cholic acid was selected as the internal standard. This newly developed method was fully validated for linearity, precision, accuracy, and stability, and then applied to quality assessment of GFC. Finally, the batch-to-batch reproducibility of GFC samples was evaluated by the cosine ration and Euclidean distance method, which showed high quality consistency. The results demonstrated that the developed method provided a reasonable and powerful manner for quality control of GFC.
基金supported by National Science and Technology Major Projects of China(2013ZX09402203,2013ZX09508104)Medical and Health Science and Technology Innovation Engineering of Chinese Academy of Medical Sciences(2016-I2M-3-007)National Natural Science Foundation of China(81573454)
文摘OBJECTIVE To evaluate the effect of Guizhi Fuling Capsule active pharmaceutical ingredient(API)and its fractions on human breast cancer cells proliferation by high-throughput screening assay.METHODS The crude fractions were obtained from the extraction and elution of the API of Guizhi Fuling Capsule,and 929 standard fractions were obtained by the optimal separation conditions.Sulforhodamine B(SRB)method was used to evaluate the effects of the Guizhi Fuling capsule API and929 kinds of fractions on the proliferation of human breast cancer cells MCF-7 and MDA-MB-231.RESULTS The Guizhi Fuling capsule API had a strong ability to inhibit the proliferation of MCF-7 cells at high concentration and the ability to inhibit MDA-MB-231 cells' proliferate at low concentration following 72 h treatment;some samples of 929 fractions(5μg·mL^(-1))was found to have a breast cancer cell growth inhibition rate above 50%,without toxicity on HUVECs proliferation.CONCLUSION The API of Guizhi Fuling capsule had significant cytotoxicity effects on these two human breast cancer cells,with significant concentration-and time-dependent manner.
基金This work was supported by the National Natural Science Foundation of China(No.82074348)the Taishan Scholar Project(tsqn201812145)the Key Technology Research and Development Program of Shandong(No.2019GSF108162).
文摘Objective To observe the effects of Guizhi Fuling capsule(GZFLC)on RPMI 8226 cells and explore the mechanisms.Methods Cell Counting Kit-8(CCK-8)assays and flow cytometry were used to detect the viability and apoptosis levels of RPMI 8226 cells.The effects on mitochondria were examined by ROS and JC-1 assays.Western blotting was used to detect the expression of B cell lymphoma-2(Bcl-2),Bax,cleaved caspase-3,and Apoptotic protease-activating factor 1(Apaf-1).Results GZFLC drug serum decreased the viability and increased the apoptosis of RPMI 8226 cells.In addition,this drug increased ROS levels and decreased the mitochondrial membrane potential(MMP).Western blotting showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups,whereas the expression levels of cleaved caspase-3 and Apaf-1 were increased.Conclusion GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway.
文摘Objective:To compare the clinical efficacy of Gongliuqing Capsule(宫瘤清胶囊)combined with mifepristone,Guizhi Fuling Capsule(桂枝茯苓胶囊)combined with mifepristone,and single mifepristone in the treatment of uterine fibroids.Methods:Retrospective analysis was performed on 307 patients with uterine fibroids receiving drug treatment in our hospital.All 307 patients has been divided into three groups according to the different drug delivery scheme,GLM group of 125 patients taking Gongliuqing Capsule and mifepristone,GFM group of 113 patients taking Guizhi Fuling Capsule(桂枝茯苓胶囊)and mifepristone,SM group of 69 patients only taking mifepristone.Comparison of the cure rate and effective rate,volume of menstruation,menstrual period and uterine fibroids,level of estrogen and the incidence of adverse reaction of the patients of the 3 groups.Results:After 90days of treatment,the efficiency of GLM and GFM group were higher than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P>0.05).The cure rate of GLM group was higher than that of the other two groups,and the difference was statistically significant(P<0.05).The menstrual volume and period length,uterine fibroids volume of GLM and GFM group was smaller than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P<0.05).The P and E2 level of GLM and GFM group was lower than that of SM group(P<0.05),and there was no significant difference between GLM and GFM group(P>0.05).There was no significant difference in the incidence of other sex hormone level and adverse reactions(P>0.05).Conclusions:Compared with the SM to treat uterine fibroids,the integrative medicine therapy were better.Among them,Gongliuqing Capsule(宫瘤清胶囊)combined with mifepristone has the highest cure rate.
基金Supported by the National Natural Science Foundation of China(No.82074348)the Taishan Scholar Project(No.tsqn201812145)+1 种基金the Key Technology Research and Development Program of Shandong(No.2019GSF108162)the Natural Science Foundation of Shandong Province(No.ZR2020MH388)。
文摘Objective: To observe the effects of Guizhi Fuling Capsule(GZFLC) on myeloma cells and explore the mechanisms. Methods: MM1S and RPMI 8226 cells were co-cultured with different concentrations of serum and the cell experiments were divided into negative(10%, 20% and 40%) groups, GZFLC(10%, 20%, and 40%)groups and a control group. Cell counting kit-8(CCK-8) assays and flow cytometry were used to detect the viability and apoptosis levels of myeloma cells. The effects on mitochondria were examined by reactive oxygen specie(ROS) and tetrechloro-tetraethylbenzimidazol carbocyanine iodide(JC-1) assays. Western blot was used to detect the expression of B cell lymphoma-2(Bcl-2), Bcl-2-associated X(Bax), cleaved caspase-3,-9, cytochrome C(Cytc)and apoptotic protease-activating factor 1(Apaf-1). RPMI 8226 cells(2×107) were subcutaneously inoculated into 48 nude mice to study the in vivo antitumor effects of GZFLC. The mice were randomly divided into four groups using a completely randomized design, the high-, medium-, or low-dose GZFLC(840, 420, or 210 mg/kg per day,respectively) or an equal volume of distilled water, administered daily for 15 days. The tumor volume changes in and survival times of the mice in the GZFLC-administered groups and a control group were observed. Cytc and Apaf-1 expression levels were detected by immunohistochemistry. Results: GZFLC drug serum decreased the viability and increased the apoptosis of myeloam cells(P<0.05). In addition, this drug increased the ROS levels and decreased the mitochondrial membrane potential(P<0.01). Western blot showed that the Bcl-2/Bax ratios were decreased in the GZFLC drug serum-treated groups, whereas the expression levels of cleaved caspase-3,-9,Cytc and Apaf-1 were increased(all P<0.01). Over time, the myeloma tumor volumes of the mice in the GZFLCadministered groups decreased, and survival time of the mice in the GZFLC-administered groups were longer than that of the mice in the control group. Immunohistochemical analysis of tumor tissues from the mice in the GZFLCadministered groups revealed that the Cytc and Apaf-1 expression levels were increased(P<0.05). Conclusion:GZFLC promoted apoptosis of myeloma cells through the mitochondrial apoptosis pathway and significantly reduced the tumor volumes in mice with myeloma, which prolonged the survival times of the mice.
基金supported by the National Natural Science Foundation of China (No.82074076)the Natural Science Foundation of Henan Province (No.202300410022).
文摘Purpose:The well-known traditional Chinese formula Guizhi Fuling capsule(GFC)has been reported to reverse ovarian cancer drug resistance.Extrachromosomal DNA(ecDNA)plays an important role in tumour metastasis and resistance.The purpose of this study was to investigate the potential mechanisms by which GFC blocks tumour metastasis and reverses drug resistance by targeting ecDNA.Methods:CNKI and PubMed were used to obtain pharmacokinetic research data on GFC in rats,and the bioactive ingredients detected in rat serum or plasma were collected.Network databases were used to screen the abnormally expressed genes in ecDNA,tumour metastasis genes,resistance genes,and the active ingredient targets of GFC.The KOBAS3.0 database was used to enrich the KEGG pathways and GO functions;the STRING platform was used to construct the core protein interaction network;and the molecular docking online tool SwissDock was used to analyse the binding activity of the core targets and the active ingredients.RT-qPCR,Western blotting and laser confocal microscopy were used to verify the efect of the sera containing GFC on ecDNA,mRNA and protein expression of key targets.Results:Twenty-three bioactive ingredients of GFC were retrieved from PubMed and CNKI.Nine shared targets were simultaneously involved in abnormal genes in ecDNA,tumour metastasis and resistance and the active ingredient targets of GFC.GO functional analysis indicated that the cotargets involved cell proliferation,apoptotic regulation,nuclear functions,etc.The potential pathways involved in the reversal of tumour metastasis and drug resistance of GFC were the PI3K-Akt signalling,cancer,and platinum drug resistance pathways.Three shared proteins targeting ecDNA(AKT1,EGFR and MYC)stand out from the top 20 PPI targets,and all of the bioactive ingredients of GFC have strong binding afnity to the three proteins.The active ingredients can reduce the expression of MYC,EGFR and AKT1 mRNA and protein and the amount of ecDNA in drug-resistant OC cells.Conclusions:GFC targeting ecDNA to reverse tumour metastasis and drug resistance has the characteristics of multiple ingredients,multiple targets,and multiple pathways,which provides a new perspective for the development of new drugs targeting ecDNA to beneft tumour treatment.
