Fuzheng Qingjie recipe induces apoptosis in HepG2 cells via P38 MAPK activation and the mitochondria-dependent apoptotic pathway

Abstract

Fuzheng Qingjie (FZQJ) recipe is a polyherbal Chinese medicine capable of suppressing tumor growth and is used as an adjuvant therapy for various types of cancer. However, its anticancer mechanisms are yet to be fully elucidated. In the present study, we explored whether p38 mitogen-activated protein kinase (MAPK) was involved in FZQJ-mediated mitochondria-dependent apoptosis in human hepatocellular carcinoma cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were used to measure the viability of HepG2 cells. 4,6-Diamidino-2-phenylindole (DAPI) and Annexin-V fluorescein isothiocyanate (FITC) were used to analyze the apoptosis of HepG2 cells. The mitochondrial membrane potential (∆ψ) and phosphorylated P38 MAPK protein were examined by a flow cytometer following 5,5',6,6'-tetrachloro‑1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1) and Alexa Fluor® 647 mouse anti-phosphorylated P38 MAPK antibody staining, respectively. The activation of caspase-9 and caspase-3 were measured using colorimetric assays. Additionally, Bcl-2 and Bax expression were examined using reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis. The results demonstrated that water extract of FZQJ was able to induce apoptosis of HepG2 cells in vitro. FZQJ-induced apoptosis was accompanied by the loss of ∆ψ, downregulation of Bcl-2 and upregulation of Bax expression, and the activation of caspase-3, -9 and P38 MAPK. These results indicated that FZQJ induced apoptosis in HepG2 cells at least via P38 MAPK activation and the mitochondria-dependent apoptotic pathway.

Fuzheng Qingjie granules potentiate the anticancer effect of cyclophosphamide by regulating cellular immune function and inducing apoptosis in Hepatoma 22 tumor-bearing mice

“…However the mechanism underlying the apoptosis-inducing action of FZQJ remains to be elucidated. Previous studies have demonstrated that FZQJ-induced hepatoma cell apoptosis occurs via the regulation of Bcl-2 and Bax expression in vitro (13). In the present study, FZQJ was observed to stimulate IL-2 and TNF-α production.…”

“…Fuzheng Qingjie granules (FZQJ) are composed of Fuzheng Guben and Qingre Jiedu herbs (13). The four Fuzheng Guben herbs are Astragalus membranaceus, Ligustrum lucidum, Ganoderma lucidum and Rhizoma dioscoreae, and the two Qingre Jiedu herbs are Hedyotis diffusa Willd and Prunella vulgaris (13).…”

“…The four Fuzheng Guben herbs are Astragalus membranaceus, Ligustrum lucidum, Ganoderma lucidum and Rhizoma dioscoreae, and the two Qingre Jiedu herbs are Hedyotis diffusa Willd and Prunella vulgaris (13). FZQJ granules are used to treat the symptoms that are commonly observed post-chemotherapy or radiotherapy, including thirst, night sweats, constipation, insomnia, loss of appetite and weakness.…”

“…FZQJ granules are used to treat the symptoms that are commonly observed post-chemotherapy or radiotherapy, including thirst, night sweats, constipation, insomnia, loss of appetite and weakness. A previous study demonstrated that FZQJ may induce apoptosis of HepG2 cells via activating p38 mitogen-activated protein kinases (MAPKs) and inducing mitochondria-dependent apoptosis (13).…”

Anticancer activities and mechanisms of heat-clearing and detoxicating traditional Chinese herbal medicine

“…Hara, Ranunculus ternatus, Pinelliae Rhizoma, Cremastrae Pseudobulbus, Arisaematis Rhizoma Preparatum, Houttuyniae Herba, and Prunellae Spica Lung cancer (LLC)Inhibiting proliferation, intervening immune escape↓CD4+CD2+Tr, ↓VEGF, ↓Scd44V6, ↓TGR-β1, ↓IL-10[102, 103]YiQi ChuTan Recipe Panacis Quinquefolii Radix , Ophiopogonis Radix, Phellodendri Chinensis Cortex, Cremastrae Pseudobulbus, Stephaniae Tetrandrae Radix , Pinelliae Rhizoma, Gynostemma pentaphyllum (Thunb.) Makino, and Hominis Placenta Lung cancer (A549, LLC)Inhibiting proliferation and metastasis, reversing EMT↓GRP78, ↓smad2/3, ↓SRC/MAPK, ↑Caspase-4, ↑DNA-PK, ↓Hspd1, ↓PH, ↓PDI, ↓EG433182, ↓HSPA 5 precursor, ↓HSPA 9, ↓PP1, ↓PRDX-1, ↓PRDX-6[104–106]Jianpi Yangzheng Xiaozheng Recipe Codonopsis Radix , Atractylodis Macrocephalae Rhizoma , Poria, Dioscoreae Rhizoma , Coicis semen, Citri Reticulatae Pericarpium , Aucklandiae Radix , Angelicae Sinensis Radix , Paeoniae Radix Alba , Smilacis Chinae Rhizoma, Salivae Chinensis Herba, and Glycyrrhizae Radix Rhizoma Praeparata Cum Melle Gastric cancer (MGC-803)Inducing apoptosis and autophagy↑Bax, ↓Bcl-2, ↓cyclin D1, ↓cyclin D2, ↓cyclin D3, ↑Fas, ↓procaspase-3, ↓procaspase-8, ↓procaspase-9, ↑cleaved-PARP, ↑Beclin-1, ↑LC3 II[107]Fuzheng Qingjie Recipe Astragali Radix, Ligustri Lucidi Fructus, Ganoderma, Dioscoreae Rhizoma, Prunellae Spica and Hedyotis diffusa Willd Hepatocellular carcinoma (HepG2)Inducing apoptosis↑caspase-9, ↑caspase-3, ↑P38 MAPK, ↑Bax, ↓Bcl-2[108]Baihe Recipe Solanum lyratum Thunb., Hedyotis diffusa Willd, Agrimoniae Herba , Codonopsis Radix, and Poria, Gastric cancer (BGC-823)Inhibiting proliferation and metastasis↓VEGF, ↓p53[109]Weikangfu Granule Curcumae Radix , Astragali Radix , Glycyrrhizae Radix Et Rhizoma, and Poria Sarcoma (S180)Inducing apoptosis and cell cycle arrest at G0/G1 phase, modulating immune response↑p53, ↑Bax, ↓Bcl-2[110, 111] …”