ABSTRACT
Objective To investigate the effects of macrophage inflammatory protein-1α (MIP-1α) combined with molecule 4-1BB L on the tumorigenicity of hepatocellular carcinoma cells in vivo. Methods Mouse MIP-1α (mMIP-1α) expressed Hepa 1-6 cells were transfected with m4-1BBL recombinant retrovirus, the anti-histidinol cells clones were selected and amplified. The expression of m4-1BB L was confirmed by flow cytometry. The growth curve of Hepa 1-6 cells transfected with mMIP-1α and m4-1BBL alone or together was drawn and compared. C57B/L Mice were randomly divided into 7 groups, 9 mice in each group, injected with mMIP-1α+m4-1BB L Hepa 1-6 cells, m4-1BB L Hepa 1-6 cells, mMIP-1α Hepa 1-6 cells, Hepa 1-6 cells, pLXSHD Hepa 1-6 cells or PBS respectively. The tumorigenicity of hepatocellular carcinoma cells and the mice survival rate were compared between each groups. Results Hepa 1-6 mMIP-1α+m4-1BB L cells which expressed both mMIP-1α and m4-1BB L were successfully established. The expression of mMIP-1α and m4-1BB L alone or together did not affect the growth curve of Hepa 1-6 cells. Observed for 5 weeks, no tumor developed in Hepa 1-6 mMIP-1α+m4-1BB L injected mice. The tumorigenicity of Hepa 1-6 mMIP-1α+m4-1BB L was lower than that of Hepa 1-6 mMIP-1α or Hepa 1-6 m4-1BB L in vivo. The survival rate of Hepa 1-6 mMIP-1α+m4-1BBL injected mice(9/9) was higher than that of Hepa 1-6 m4-1BB L injected mice (6/9)or Hepa 1-6 mMIP-1α injected mice (1/9). Conclusion Chemokine MIP-1α combined with costimulatory 4-1BB L lowered the tumorigenicity of hepatocellular carcinoma cells in vivo, and prolonged the mice survival period.
ABSTRACT
Objective: To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages. Methods: LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay. Results: Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels. Conclusion: In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.
ABSTRACT
Objective To investigate hypolipemic treatment of hyperlipidemic panereatiti(HLP)with integrated traditional Chinese and western medicine.Methods Cinical data of 20 patients of HLP were analyzed retrospectively.Eight patients in the control group were treated with conventional therapies,while 12 patients in the treatment group were treated as follows:①Enema with 180ml solution(50% magnesium sulfate 30 ml.Glycerin 60ml,water 90ml).②Rhubarb gastrogavage with 9 g tid.③Intravenous drip with 24 g salvia miltiorrhiza qd.Results The treatment group had significant difference comparing with the control group in terms of the serum TG in 48 hours(P<0.01),time of autonomous bowel movement recover(P<0.01),days of abdominal pain disappear(P<0.05),days of hospitalization(P<0.01).Conclusion The treatment of Enema with 180 ml solution.Rhubarb gastrogavage with 9g tid,and Intravenous drip with 24 g salvia miltiorrhiza qd can relieve the symptoms of HLP and decrease blood-fat greatly.
ABSTRACT
Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP-1α and to evaluate the possibility of cancer gene therapy by mMIP-1α. Methods: mMIP-1α cDNA was cloned into retrovirus vector pBabe puro and pBabe puro-mMIP-1α was constructed, then pBabe puro-mMIP-1α was used to transfect packaging cells, anti-puromycin cells was proliferated, the supernatant was used to infect hepa1-6, the anti-puromycin clone (hepa1-6 mMIP-1α) and hepa1-6 were analysed for the expression of mMIP-1α mRNA and protein by RT-PCR and immunohistochemistry respectively. The growth curve of hepa1-6 and hepa1-6 mMIP-1α was drawn. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro-mMIP-1α with mMIP-1α cDNA was constructed. Hepa1-6 did not produce mMIP-1α mRNA and protein, while hepa1-6 mMIP-1α could produce mMIP-1α mRNA and protein. The growth curve of hepa1-6 and hepa1-6 mMIP-1α showed no difference. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1-6 mMIP-1α is established and mMIP-1α can affect the tumorigenecity of hepa1-6.
ABSTRACT
Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP 1? and to evaluate the possibility of cancer gene therapy by mMIP 1?. Methods: mMIP 1? cDNA was cloned into retrovirus vector pBabe puro and pBabe puro mMIP 1? was constructed, then pBabe puro mMIP 1? was used to transfect packaging cells, anti puromycin cells was proliferated, the supernatant was used to infect hepa1 6, the anti puromycin clone (hepa1 6 mMIP 1?) and hepa1 6 were analysed for the expression of mMIP 1? mRNA and protein by RT PCR and immunohistochemistry respectively. The growth curve of hepa1 6 and hepa1 6 mMIP 1? was drawn. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro mMIP 1? with mMIP 1? cDNA was constructed. Hepa1 6 did not produce mMIP 1? mRNA and protein, while hepa1 6 mMIP 1? could produce mMIP 1? mRNA and protein. The growth curve of hepa1 6 and hepa1 6 mMIP 1? showed no difference. The chemotaxis of mMIP 1? produced by hepa1 6 mMIP 1? to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1 6 mMIP 1? is established and mMIP 1? can affect the tumorigenecity of hepa1 6.