ABSTRACT
OBJECTIVE: To evaluate the apoptosis effect of HDAC inhibitor trichostatin A (TSA) combination with Hedgehog (Hh) signaling inhibitor cyclopamine (CYA) on the PANC-1 cells line. METHODS: Hochest 33258 staining was used to measure apoptosis of PANC-1 cells treated with TSA and CYA respectively or cooperatively. The 24 h IC50 of PANC-1 cells line responding to TSA and CYA were detected by CCK8. The mRNA expression of the gene relative with apoptosis and Hh signaling were evaluated by SYBR GREEN based FQ-PCR (fluorescent quantization polymerase chain reaction). The proteins associated with apoptosis (Bcl-2 and activated caspase-3, activated caspase-8, activated caspase-9) and Hh signalling pathway (Gli1, Smo and Ptc-1) were measured by Western blotting (WB). Gli1 was also detected by cell immunofluorescence. RESULTS: TSA and CYA could induce apoptosis of PANC-1 cells by Hochest 33258 stain, respectively or combinatively. The 24 h IC50 of PANC-1 cells line treated with chemical were (0.51±0.07) μmol·L-1 (TSA) and (33.6±2.3) μmol·L-1 (CYA), respectively; the combination index (CI) of TSA and CYA on the PANC-1 is 0.835. Combination usage of TSA and CYA caused more significantly decreased expression of Bcl-2 mRNA and increased Bax mRNA as compared with the TSA or CYA treatment. TSA (0.5 μmol·L-1) inhibited the expression of Gli1 mRNA, TSA promoted the inhibition effect of CYA on Hh pathway, down-regulation of Gli1, Smo mRNA and up-regulation of Gli3 mRNA. Western blotting showed that combination usage of TSA and CYA induced more significant activation of activated caspase-3, activated caspase-8, activated caspase-9 and reduction of Bcl-2, Gli1, Smo and Ptc-1 expression than TSA or CYA treatment. Decreased Gli 1 also confirmed by immunofluorescence. CONCLUSION: Combination usage of TSA and CYA could induce PANC-1 cells line apoptosis and synergistically inhibit Hh pathway.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the efficacy and feasibility of duodenojejunal bypass(DJB)on non-severe obese patients with type 2 diabetes mellitus(T2DM).</p><p><b>METHODS</b>The body mass index (BMI), fasting plasma glucose(FPG), 2h-postprandial plasma glucose(2hPG), fasting insulin(F-ins), fasting c-peptide(F-CP), glycated hemoglobin and hypoglycemic agents dose changes were tested in 7 patients with non-severe obese T2DM undergoing DJB, preoperatively and within 24 weeks after surgery during the follow-up. Data were collected and the clinical outcomes of T2DM were analyzed.</p><p><b>RESULTS</b>In 7 cases of non-obese T2DM who underwent DJB, one patient was weaned off hypoglycemic agents with normal FPG, 2hPG and HbA1c postoperatively. Five required significantly lower dosage. No significant improvement in 1 case. Complete remission rate of hyperglycemia was 1/7, effective rate was 6/7, and effective rate of HbA1c was 5/7. No significant changes in BMI were observed between the preoperative and postoperative phases.</p><p><b>CONCLUSION</b>Plasma glucose level can be markedly reduced by duodenojejunal bypass in non-obese T2DM, independent of weight loss, and the mechanism remains unclear.</p>