[Effect of complete Freund's adjuvant on islet beta cell apoptosis and its mechanism in non-obese diabetic mice].
OBJECTIVE
To investigate the effect of complete Freund's adjuvant (CFA) on islet beta cell apoptosis in preventing diabetes in non-obese diabetic (NOD) mice, and the influence on apoptotic related-gene expression.


METHODS
Four-week-old female NOD mice were randomly divided into Group CFA (n=5) and Group saline (NS) control (n=5). Mice in Group CFA were injected in the hind footpad with 50 microL CFA and mice in Group NS with 50 microL NS. Blood sugar was monitored and diabetes was diagnosed if blood sugar was higher than 11.1 mmol/L for 2 continuous days in the NOD mice. The mice were sacrificed when diagnosed as diabetes or at 30 weeks of age. Pancreatic sections were made for: evaluation of insulitis severity with HE staining; counting of apoptotic beta cells with the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method, and ABC immunohistochemical double labeling; counting of Fas, FasL and Bcl-x positive cells respectively with ABC immunohistochemical method.


RESULTS
By 30 weeks of age, none of the 5 CFA-treated mice, compared with 3 of the 5 control mice, had developed diabetes. The insulitis score was lower (1.820+/-0.962 vs. 3.020+/-1.040, P<0.05), the rates of apoptotic beta cells, Fas positive cells and FasL positive cells were lower [(10.2+/-2.8)% vs. (15.9+/-6.5)%, (54.9+/-14.5)% vs. (75.7+/-12.9)%, (20.3+/-10.4)% vs. (27.9+/-12.0)%, P<0.05), and the Bcl-x positive cell rate was higher [(74.9+/-10.7)% vs. (66.0+/-18.3)%, P<0.05] in the CFA-treated group than those in the NS-treated group respectively.


CONCLUSIONS
CFA may inhibit beta cell apoptosis in NOD mice by regulating Fas, FasL and Bcl-x expression on cells within islets.
Zhu
Lin
L
Diabetes Center, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Zhou
Zhi-guang
ZG
Yang
Wen
W
Yang
Zhu-lin
ZL
Zhang
Song
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D003922:Diabetes Mellitus, Type 1 / Q000473:pathology*; D053222:Fas Ligand Protein / Q000378:metabolism; D005260:Female; D005620:Freund's Adjuvant / Q000494:pharmacology*; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000473:pathology*; D051379:Mice; D016688:Mice, Inbred NOD; D051020:bcl-X Protein / Q000378:metabolism; D019014:fas Receptor / Q000378:metabolism
C494812:Bcl2l1 protein, mouse; D053222:Fas Ligand Protein; C491899:Fas protein, mouse; C505658:Fasl protein, mouse; D051020:bcl-X Protein; D019014:fas Receptor; D005620:Freund's Adjuvant