Toxicity study on bupivicaine to SH-SY5Y cells cultured by high-glucose / 重庆医学

Objective To discuss the effect of bupivacaine on SH‐SY5Y cells in high sugar environment and observe the ROS and apoptosis .Methods 1 mmol/L bupivicaine was added to medium with different concentrations of glucose for SH‐SY5Y cells , and the quantity of ROS in the cells and the situation of apoptosis were detected by flow cytometry ,and Western blot was uesd to detect the change of the related protein ,GRP78 .Results The contents of intarcellular ROS in different groups ,which had different concentration(7 .80 ,11 .10 ,13 .30 mmol/L) of sugar medium ,were high than those in groups with concentrations 5 .56 ,6 .10 ,7 .00 mmol/L of sugar medium ,and it showed statistical significance (P<0 .05) .Cell apoptosis rates among different groups had statisti‐cal significance(P<0 .05) .Expression of GRP78 in group of 5 .56 ,6 .10 and 7 .80 mmol/L were (1 .02 ± 0 .12) ,(0 .97 ± 0 .06) and (0 .49 ± 0 .04) ,respectively ,and they all were higher than that in group of 7 .00 mmol/L (0 .46 ± 0 .06) .The groups of 11 .10 mmol/L (0 .22 ± 0 .03) and 13 .30 mmol/L (0 .15 ± 0 .07) were lower than the other groups in the expression of GRP78 ,and all the difference shows statistical significance (P<0 .05) .Conclusion Increasing and apoptosis of intracellular ROS are associated with strengthened endoplasmic reticulum stress by bupivacaine .With the function degrading of GRP78 ,endoplasmic reticulum stress (ERS) may strengthen further .

To constructe,package and identificate the lentiviral vector with overexpression gene Grp78 / 重庆医学

Objective To constructe ,package and identificate the lentiviral vector with overexpression gene Grp78 .Methods We used lentiviral vector and genetic engineering technology to obtain the aim gene fragment and to constructe recombinant plas‐mid .we prepared competent cells and transform the cells .Through positive clone sequencing ,lentivirus was packaged and virus titer was tested .Results Positive cloning sequence comparison results show that the test was passed .Melt curve did not appear mixed peak ,also did not appear abnormal peak broadening .It means that does not appear pollution ,primer dimers and nonspecific amplifi‐cation in the experiments .Conclusion The construction ,packaging and identification of lentiviral vector with over expression gene Grp78 are sucessful .