ABSTRACT
Objective To establish the Nifedipine-induced liver cell damage model,and to investigate the cellular or molecular mechanism for children's liver cell damage.Methods The HepG2 cells were utilized to establish liver cell damage models.The optimal concentration and the optimal pretreatment time of Nifedipine-induced liver cell injury were confirmed.Western blot and real-time PCR(RT-PCR)were used to check the alteration in proteins and mRNAs level of the liver function-associated classic markers,which contained alkaline phosphatase(ALP),aspartate amino transferase(AST),glutamyl transpeptidase(γ-GT)and alanine aminotransferase(ALT).Additionally,flow cytometry(FCM),colony formation assay(CFA)and cell wound healing assay(CWHA)were utilized to check the effect of Nifedipine on the cell cycle progression and proliferation of HepG2.Results (1)The optimal concentration of Nifedipine was 20 mg/L and the optimal treatment period was 21 days for liver damage.(2)Western blot:intracellular ALT protein content after Nifedipine group was less than that of control group,while the protein levels of AST,γ-GT and ALP in the culture medium after Nifedipine addition(3.55 ± 0.05,4.91 ± 0.055,3.51 ± 0.05,3.08 ± 0.08) were higher than those of control group(0.96 ± 0.02,1.03 ± 0.02,1.00 ± 0.05,0.90 ± 0.13),and the differences were all significant(t = -85.695,-117.582,-47.371,-33.260,all P<0.05).(3)The findings of RT-PCR showed that the mRNA levels of intracellular ALT,γ-GT and ALP in Nifedipine group(0.26 ± 0.02,0.05 ± 0.04, 0.05 ± 0.02)were lower than those of control group(1.13 ± 0.21,0.94 ± 0.10,1.03 ± 0.06),and the differences were all significant(t=7.233,127.436,25.687,all P<0.05).However,the mRNAs levels in purified culture me-dium in Nifedipine group(5.95 ± 0.05,3.13 ± 0.10,3.32 ± 0.08)were higher than those in control group(1.01 ± 0.08,1.00 ± 0.05,1.00 ± 0.05),and the differences were all significant(t= -92.339,-31.250,-43.007,all P<0.05).(4)The portion of G0/G1 phase in Nifedipine group[(84.09 ± 0.43)%]was more than that of control group[(30.93 ± 0.32)%],which had statistical significance(t=173.084,P=0.000).(5)In contrast with control group,the colony formation of cells in Nifedipine group declined from(97.10 ± 1.17)% to(38.56 ± 1.51)%(t=92.088,P=0.000)and the migration rate of cells wound healing was(56.37 ± 2.06)%,(25.00 ± 1.71)% sepa-rately(t=20.285,P=0.000).Conclusion Nifedipine may promote children's liver injury through regulating cell cycle related proteins.
ABSTRACT
Background:The incidence of chronic constipation is increasing in recent years,which seriously affects the quality of life of patients. Aims:To investigate the therapeutic efficacy of lactulose combined with Bacillus subtilis and Enterococcus faecium preparation on chronic constipation. Methods:A total of 112 patients with chronic constipation from Jan. 2011 to Jan. 2014 at Emeishan Hospital of TCM were enrolled,and were randomly divided into observation group and control group. Patients in both groups accepted the guidance of diet. In addition,patients in control group were orally administrated with lactulose while patients in observation group were orally administrated with lactulose combined with Bacillus subtilis and Enterococcus faecium preparation. After 4 weeks of treatment,therapeutic efficacy in the two groups was compared,patient assessment of constipation quality of life scale(PAC-QOL)was used to evaluate the quality of life. Results:The overall therapeutic efficacy rate in observation group was significantly higher than that in control group (98. 2% vs. 72. 2% ;χ2 = 12. 399,P < 0. 05). Score of every factor and the total score of PAC-QOL were significantly lower after treatment than those before treatment in both groups(P < 0. 05),while physical discomfort score and the total score in observation group were significantly lower than those in control group( P < 0. 05). Conclusions:Lactulose combined with Bacillus subtilis and Enterococcus faecium preparation can effectively improve the symptoms of difficulty in defecation,hard stool,and improve the quality of life in patients with chronic constipation.