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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.
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<p><b>OBJECTIVE</b>To further investigate the {ptin vitro} effects of an osteoprotective herbal formula "ELP" (Herba Epimedii, Fructus Ligustri Lucidi and Fructus Psoraleae) using seropharmacological approach.</p><p><b>METHODS</b>Rats were fed with ELP or its individual component herbs for 2 days. The serum containing the postabsorbed ingredients of the herbal items were collected for cell culture using UMR106 cell, RAW264.7 cell and mesenchymal stem cell (MSC) isolated from the bone marrow of the rats. The effects of the herbal-containing serum on cell toxicity were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay; bromodeoxyuridine assay was conducted to measure the cell proliferation of UMR106 cell and MSC; cell activity was measured using colorimetric method, and mRNA expression of runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteopontin (OPN) of UMR106 and MSC as well as matrix metalloproteinase 9 (MMP-9), tartrate-resistant acid phosphatase (TRAP) and cathepsin K of RAW264.7 were analyzed using real-time reverse-transcription polymerase chain reaction.</p><p><b>RESULTS</b>ELP and its component serum exhibited no cytotoxic effects on the cells. The ELP-containing serum increased the proliferation of UMR106 cell and MSC by 25.7% and 14.4 %, respectively and the alkaline phosphatase activity of MSC was increased by 42.6%. On the contrary, it inhibited the RAW264.7 cell differentiation by 29.2 %. ELP serum upregulated the Runx2 expression of UMR and MSC by 1.18 fold and 1.27 fold, respectively. It also upregulated ALP and OPN expression in MSC by 1.69- and 2.12-fold, respectively. On the other hand, ELP serum down-regulated MMP-9 and cathepsin K expression of RAW264.7 cell by 0.46- and 0.36-fold, respectively.</p><p><b>CONCLUSIONS</b>The serum of the animals fed with ELP contains active ingredients which are effective in promoting osteogenesis and inhibiting osteoclastogenesis.</p>
Subject(s)
Animals , Male , Mice , Rats , Absorption, Physiological , Bone and Bones , Pathology , Cell Differentiation , Cell Proliferation , Cell Survival , Drugs, Chinese Herbal , Pharmacology , Osteoclasts , Metabolism , Pathology , Osteogenesis , Protective Agents , Pharmacology , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Serum , MetabolismABSTRACT
Objective To investigate the influence of serum containing Qingre Chubi Decoction ( QCD) on the THP-1 cell viability and the release of interleukin 1 beta ( IL-1β) stimulated by monosodium urate crystals in vitro. Methods The cultured human monocyte THP-1 strain were divided into blank serum group, model control group, and high-, middle- and low-concentration ( volume fraction being 20%, 10%, 5%) QCD-containing serum groups. Except for the blank serum group , the other groups were all given 500 mg/L of monosodium urate crystals. On culturing hour 0, 12, 24 and 48, THP-1 cell viability was tested by methy1 thiazolyl tetrazolium celorimetry ( MTS) method. On culturing hour 48, the content of IL-1β in the supernatant of THP-1 cells was detected by enzyme-linked immunosorbent assay ( ELISA) . Results The THP-1 cell viability in various groups was increased along with the prolongation of culturing time. The THP-1 cell viability in the model control group was increased as compared with that in the blank serum group at different time points (P<0.05 or P<0.01) . And the content of IL-1β in the model control group was increased significantly as compared with that in the blank serum group on culturing hour 48 (P<0.01) . The THP-1 cell viability in various QCD-containing serum groups on culturing hour 12 and 24, and in high- and middle-concentration QCD-containing serum group on culturing hour 48 was decreased significantly as compared with that of the model control group at the same time point ( P<0.05 or P<0.01) . The content of IL-1β in various serum containing QCD groups was markedly decreased as compared with that in the model control group on culturing hour 48 ( P<0.01) . Conclusion Serum containing QCD can inhibit the viability of THP-1 cells stimulated by monosodium urate crystals, and the possible mechanism is related with the inhibition of IL-1 release.
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【Objective】To investigate the effect of serum containing Yifa Compound(YC) on the growth of hair papilla cells.【Methods】Hair papilla cells were cultured in vitro.After subculture for 4～6 generations,hair papilla cells at logarithmic growth phase were cultured with YC-containing serum(at the concentrations of 1.25,2.5 and 5.0?g/L respectively) and blank serum respectively.After the culturing,the proliferation curve of hair papilla cells was drafted by methylthiazolyltetrazolium(MTT) assay and their growth was observed under scanning electron microscope.【Results】Optical density of hair papilla cells was higher at logarithmic growth phase while lower at growth lag phase in high-dose YC-containing serum group than that in blank serum group.Moderate-and high-dose YC-containing serum promoted the agglutinating radial growth.【Conclusion】Serum containing YC can promote the proliferation of hair papilla cells.
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OBJECTIVE:To study the antitumor mechanism of Ruixiang Langdu(Stellera chamaejasme L ) abstracts(SCA) METHODS:SCA-contained serum was derived from mice pre-administrated with different oral dosages of SCA and at different times after administration The effects of the serum on the proliferation of K562 leukemic cells were observed with MTT assay and clone formation RESULTS:After mixing with the SCA-contained serum derived at 1,2,4,8h after giving SCA(3,6 and 12g/kg),the rates of MTT transformation and clone formation of K562 cells were decreased significantly The SCA-contained serum 12 h after giving drug was more effective than others CONCLUSION:The SCA-contained serum inhibited the proliferation of tumor cells,which may be one of its important antitumor mechanism