Jiedu Recipe, a compound Chinese herbal medicine, inhibits cancer stemness in hepatocellular carcinoma via Wnt/β-catenin pathway under hypoxia / 中西医结合学报

OBJECTIVE@#Jiedu Recipe (JR), a Chinese herbal remedy, has been shown to prolong overall survival time and decrease recurrence and metastasis rates in patients with hepatocellular carcinoma (HCC). This work investigated the mechanism of JR in HCC treatment.@*METHODS@#The chemical constituents of JR were detected using liquid chromatography-mass spectrometry. The potential anti-HCC mechanism of JR was screened using network pharmacology and messenger ribonucleic acid (mRNA) microarray chip assay, followed by experimental validation in human HCC cells (SMMC-7721 and Huh7) in vitro and a nude mouse subcutaneous transplantation model of HCC in vivo. HCC cell characteristics of proliferation, migration and invasion under hypoxic setting were investigated using thiazolyl blue tetrazolium bromide, wound healing and Transwell assays, respectively. Image-iT™ Hypoxia Reagent was added to reveal hypoxic conditions. Stem cell sphere formation assay was used to detect the stemness. Epithelial-mesenchymal transition (EMT) markers like E-cadherin, vimentin and α-smooth muscle actin, and pluripotent transcription factors including nanog homeobox, octamer-binding transcription factor 4, and sex-determining region Y box protein 2 were analyzed using Western blotting and real-time polymerase chain reaction. Western blot was performed to ascertain the anti-HCC effect of JR under hypoxia involving the Wnt/β-catenin pathway.@*RESULTS@#According to network pharmacology and mRNA microarray chip analysis, JR may potentially act on hypoxia and inhibit the Wnt/β-catenin pathway. In vitro and in vivo experiments showed that JR significantly decreased hypoxia, and suppressed HCC cell features of proliferation, migration and invasion; furthermore, the hypoxia-induced increases in EMT and stemness marker expression in HCC cells were inhibited by JR. Results based on the co-administration of JR and an agonist (LiCl) or inhibitor (IWR-1-endo) verified that JR suppressed HCC cancer stem-like properties under hypoxia by blocking the Wnt/β-catenin pathway.@*CONCLUSION@#JR exerts potent anti-HCC effects by inhibiting cancer stemness via abating the Wnt/β-catenin pathway under hypoxic conditions. Please cite this article as: Guo BJ, Ruan Y, Wang YJ, Xiao CL, Zhong ZP, Cheng BB, Du J, Li B, Gu W, Yin ZF. Jiedu Recipe, a compound Chinese herbal medicine, inhibits cancer stemness in hepatocellular carcinoma via Wnt/β-catenin pathway under hypoxia. J Integr Med. 2023; 21(5): 474-486.

Exploration of the therapeutic mechanism of Yiqi Jiedu recipe for treatment of primary liver cancer based on network pharmacology and molecular docking / 南方医科大学学报

OBJECTIVE@#To explore the effective components of Yiqi Jiedu recipe and the main biological processes and signal pathways involved in the therapeutic mechanism of the recipe in treatment of primary liver cancer through network pharmacology and molecular docking approaches.@*METHODS@#TCMSP, Uniport, Genecards and String databases were searched to obtain the target genes of drugs and disease using Cytoscape 3.8.2 software. GO and KEGG enrichment analyses were performed to identify the common genes in the target genes of the drugs and disease. Using Pubcham, RCSB and Autoduck, the effective components of the drugs were connected with the final core genes. The effects of different concentrations of Yiqi Jiedu recipe on the expressions of the core genes DHX9, HNRNPK, NCL and PABPC1 in HepG2 cells were analyzed with Western blotting and real- time fluorescence quantitative PCR.@*RESULTS@#We finally identified 8 core genes from the drug and disease targets, including DDX5, HNRNPK, PABPC1, DHX9, RPS3A, RPS3, RPL13, and NCL. GO analysis showed that these core genes were involved mainly in the biological processes of adrenaline receptor signal communication, movement of cellular or subcellular components, blood particles, adhesion class and iron ion binding. KEGG analysis showed that the Ras signaling pathway had the greatest gene enrichment. The results of molecular docking suggested that the effective components of the recipe were capable of docking with the core genes under natural conditions, and PABPC1 and stigmasterol had the highest binding energy. In HepG2 cells, treatment with 10% medicated serum for 48 h had the strongest effect on the expression of DHX9, HNRNPK, NCL and PABPC1 (P < 0.05).@*CONCLUSION@#Yiqi Jiedu recipe is capable of regulating viral expression of primary liver cancer multiple effective components that bind to DHX9, HNRNPK, NCL and PABPC1.

Study on protective mechanism of compatibility of Huoxue Jiedu recipe on H9C2 myocardial cell autophagy induced by hypoxia/reoxygenation / 中国药理学通报

Aim To investigate the protective effect of Huoxue Jiedu recipe on autophagy injury of H9C2 cardiomyocytes induced by hypoxia/reoxygenation and its mechanism. Methods H9C2 cardiomyocytes were used to establish a hypoxia/reoxygenation injury model. The effective concentration was screened and the cell activity was detected by CCK8 assay. The apoptotic rate of myocardial cells was detected by flow cytometry. The expression of autophagy marker LC3 was observed by laser confocal microscopy. The mRNA levels of Beclin-1, LC3 and Bcl-2 were detected by real-time quantitative PCR. The expressions of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, Bcl-2, p62, p-Akt, p-mTOR were detected by Western blot. Results Huoxue Jiedu recipe can enhance the growth activity of myocardial cells and reduce the apoptotic rate and autophagy level, and it can enhance the activation of PI3K/Akt/mTORCl pathway, decrease Beclin-1 and LC3 mRNA levels, while increase Bcl-2 mRNA levels. It also decreased the expression of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, and increased the expression of p62, p-Akt, p-mTOR, and Bcl-2. Conclusions Huoxue Jiedu recipe can reduce the level of autophagy and apoptosis of myocardial cells by regulating the autophagy pathway of PI3K/Akt/mTORCl, thereby playing a protective role in hypoxia/reoxygenation H9C2 myocardial cells.

Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells / 中国实验方剂学杂志

Objective:To observe the effect of Huayu Jiedu recipe （HYJDR） on apoptosis of gastric cancer cells， explore the mechanism of HYJDR on apoptosis of gastric cancer cells and provide experimental basis for clinical application of HYJDR. Method:Thiazolyl blue tetrazolium bromide （MTT） method was used to detect the effect of HYJDR （5，10，15，20，25，30 g·L-1） on the activity of SGC-7901 cells and the median inhibition concentration （IC50） of HYJDR on SGC-7901 cells. Propidium iodide （PI） and Hoechst double fluorescence staining were used to detect the effect of HYJDR on the apoptosis of gastric cancer SGC-7901 cells. Western blot was used to detect the effect of HYJDR on the expression of apoptosis related proteins in gastric cancer cells. Real-time polymerase chain reaction （Real-time PCR） was used to detect the effect of HYJDR on the expression of apoptosis related protein mRNA in gastric cancer cells. Result:As compared with blank group， HYJDR （>10 g·L-1） can significantly inhibit the proliferation of gastric cancer cells. With the increase of the concentration of HYJDR， the cell viability decreased significantly obviously in a concentration-dependent manner（P<0.05）. HYJDR can significantly promote the apoptosis of gastric cancer cells. With the increase of the concentration of HYJDR， the apoptosis of gastric cancer cells gradually increased. As compared with blank group， HYJDR can obviously inhibit the expression of apoptosis-related B -cell lymphoma-2 （Bcl-2） （P<0.05）， and increase the expression of apoptosis-promoting proteins such as Bcl-2 antagonist of cell death （Bad） and Bcl-2-associated X （Bax）（P<0.05）. Real-time PCR results showed that as compared with blank group， HYJDR（5，10 g·L-1） could effectively inhibit the expression of Bcl-2 protein mRNA in gastric cancer cells（P<0.05）， and all HYJDR groups could promote the expression level of Bad mRNA（P<0.05）. Conclusions:HYJDR can obviously promote the apoptosis of SGC-7901， and its effect is probably achieved by increasing the expression of Bad mRNA and inhibiting the expression of Bcl-2 protein.

Anti-tumor Effects of Yiqi Jiedu Recipe and Analysis of Its Bioinformatics / 广州中医药大学学报

Objective To observe the inhibitory effects of Yiqi Jiedu Recipe(YJR)on transplanted Lewis lung cancer in mice and to explore its relative mechanism. Methods In-vivo mouse model of transplanted Lewis lung cancer was established. Twenty modeled mice were divided into model group,low- and high-dose YJR groups, and cisplatin group, 5 mice in each group. After treatment, the mouse general situations, body mass, body mass excluding tumor mass,tumor mass,tumor index,tumor volume,and tumor-inhibition rate were detected. In the in-vitro experiments, the inhibitory effect of YJR on the proliferation of A549 and H1299 cells was evaluated by CCK8 assays, and the effect of YJR on the apoptosis of A549 and H1299 cells was evaluated by flow cytometry assay. Bioinformatics analysis was carried out to identify the critical function,pathways and genes with bioinformatics mining method. Results Compared with the model group, high-dose YJR group and cisplatin group significantly inhibited the tumor growth of the transplanted Lewis lung cancer in mice(P<0.05 or P < 0.001). The experimental results in vitro showed that YJR significantly inhibited lung cell growth, and induced lung cell apoptosis. Bioinformatics analysis results showed that the inhibitory effects of YJR was probably related to multiple paths including apoptosis,cell cycle,autophagy,etc.,and promoting apoptosis may be the important way. YJR played a role in promoting apoptosis via AKT, HSP90, BCL2, CASP9 and activation of PI3K-AKT pathway. Conclusion YJR exhibits anti-tumor actions through regulating the targets AKT,HSP90, BCL2, CASP9 and affecting PI3K-AKT pathway to promote apoptosis and inhibit cell proliferation, thus to suppress the growth of transplanted tumor.

The effect of Yiqi-jianpi-jiedu recipe decoction containing serum on oxaliplatin-induced HepG2 human hepatoma cell / 国际中医中药杂志

Objective To explore the effect of Yiqi-jianpi-jiedu recipe decoction containing serum on Oxaliplatin-induced HepG2 Human hepatoma cell survivin, caspase-3, caspase-9 expression. Methods The serum pharmacological method was used to prepare nourishing Qi and invigorating spleen decoction containing serum of rat. The HepG2 human hepatoma cells were divided into Chinese herb containing serum group, chemotherapy with Oxaliplatin (OXA) group and the combination group (medicine containing serum and OXA), and the blank group without drugs. Apply MTT assay, Western Blotting method and RT-PCR method to detect the cell inhibition rate and survivin, caspase-3, caspase-9 protein and gene expression changes. Results Compared with the blank group, the inhibition rate of HepG2 Human hepatoma cell was significantly higher in Chinese herb containing serum group, chemotherapy group and the combination group (P < 0.05). Compared with thechemotherapy group and Chinese herb containing serum group, the Survivin (0.151 ± 0.054 vs. 0.288 ± 0.089, 0.375 ± 0.063) and the expression of Survivin mRNA (0.205 ± 0.091 vs. 0.487 ± 0.073, 0.725 ± 0.092) of the conbination group downgraded significantly lower (P < 0.05 or P< 0.01). Compared with chemotherapy group and Chinese herb containing serum group, the caspase-3 (0.821 ± 0.079 vs. 0.634 ± 0.098, 0.487 ± 0.102), caspase-9 (0.901 ± 0.047 vs. 0.709 ± 0.054, 0.402 ± 0.012) expression, caspase-3 mRNA (1.928 ± 0.226 vs. 1.564 ± 0.195, 1.287 ± 0.312) and caspase-9 mRNA (2.063 ± 0.517 vs. 1.536 ± 0.084, 1.019 ± 0.182) expression of the conbination group upgraded significantly higher (P < 0.05 or P < 0.01). Conclusions Nourishing-Qi and invigorating spleen decoction containing serum may affect Oxaliplatin-induced HepG2 apoptosis mechanism through increasing the suppression on Survivin and promoting the expression of caspase-3 and caspase-9 protein and genes. This research provided important experimental basis for clinical application of traditional Chinese medicine treating liver cancer.

Effects ofYiqi Huoxue Jiedu Recipe on Immunomodulatory of Patients with Advanced Ovarian Cancer / 中国中医药信息杂志

Objective To study the effects ofYiqi Huoxue Jiedu Recipe on immunomodulatory of patients with advanced ovarian cancer.Methods It was performed in a single-arm, open-label study with 40 patients with advanced epithelial ovarian cancer. All patients were treated withYiqi Huoxue Jiedu Recipe lasting for 4 weeks (200 mL, twice a day). The main endpoint was the quantities of CD4+CD25+ cells, CD8+CD28- regulatory T cells and CD8+CD28+ cytotoxic T cells measured by flow cytometry, and the accessory endpoints were variations of symptoms which included pain, fatigue, and numbness etc.Results The quantities of CD4+CD25+ and CD8+CD28- cells significantly decreased compared with those of pre-treatment (P<0.05); the quantity of CD8+CD28+ cell increased significantly (P<0.05). Except for pain, significant improvement showed in fatigue, tinnitus, numbness, poor appetite, abdominal discomfort, insomnia, diarrhea and abnormal urinate (P<0.05).ConclusionYiqi Huoxue JieduRecipe is proved to be effective in regulating immunosuppression advanced ovarian cancer and improving patients’ life quality.

Protective effects of Yiqi Huoxue Jiedu Recipe on ischemic brain injury / 中国药学杂志

OBJECTIVE: To observe the protective effects and mechanisms of Yiqi Huoxue Jiedu Recipe traditional Chinese medicine on cerebral ischemia and reperfusion injury. METHODS: SD rats were r andomly divided into sham-operation group, model group, saponin group, flavonoids group, polysaccharide group, combination group of saponin, flavonoids and polysaccharide, and nimodipine group. Using focal middle cerebral artery embolization(MCAO) model, the symptoms of neurological deficit score were observed;the infarct volume, brain index, brain water content, SOD activity, the contents of MDA, IL-1β, IL-6 and TNF-α of ischemic brain tissue, the expressions of Bcl-2 and Bax proteins, and the quantity of nissl Body were determined. RESULTS: The combination with saponin, flavonoids and polysaccharide, the main active part of Yiqi Huoxue Jiedu Recipe, could significantly reduce the symptoms of neurological deficits, promote the recovery of neurological deficits, narrow infarct volume of brain tissue, reduce the percentage of infarct volume, brain index and brain water, raise SOD activity, reduce the contents of MDA, IL-1β, IL-6 and TNF-α of ischemic brain tissue. It also increased Bcl-2protein, reduced Bax expression and reperfusion significantly. The reduction or disappearance of nissl body was inhibited. CONCLUSION: The combination with saponin, flavonoids and polysaccharide has significant protective effects on ischemic brain injury.

Effect of Huoxue Jiedu Recipe Medicated Serum on Proliferation and Cell Cycle Status of Keratinocytes of Psoriasis / 中医杂志

Objective To observe the effect of Huoxue Jiedu Recipe(Herbal recipe for activating blood and removing toxins)on proliferation and cell cycle status of keratinocytes cultured in vitro in psoriasis.Methods Using the serum pharmacologic technique,the clean SD rats were adopted to make the Huoxue Jiedu Recipe medicated serum(high,moderate,low dose),Dexamethasone medicated serum,and blank serum,which were added into the keratinocytes culture system respectively.After incubated for 48 hours,the cell proliferation of keratinocytes was measured by MTT assay,and the cell cycle distribution estimated with flow cytometer(FCM).Results The different density of Huoxue Jiedu Recipe could all inhibit significantly the generation of COLO-16 keratinocyte line and change the distribution proportion of each phase in the cell division cycle.Conclusion Huoxue Jiedu Recipe treats psoriasis through inhibiting the proliferation of keratinocytes.