[Protective effect of recombinant adenovirus carrying rat insulin-like growth factor 1 on rat islet beta-cell against strepozotocin-induced impairment in vitro].
OBJECTIVE
To construct the recombinant adenovirus containing rat insulin-like growth factor 1 (rIGF-1), and then infect to rat islet beta cells-RINm5F cells with the virus to investigate the role of rIGF-1 to streptozotocin-induced cell impairment in vitro.


METHODS
Recombinant adenovirus encoding rIGF-1 was constructed, and then infect to RINm5F cells. rIGF-1 protein was detected by Western blot analysis and ELISA method. Then streptozotocin was used to induce RINm5F cells impairment. The levels of nitric oxide were detected in cells culture supernatants. The cells function was evaluated by glucose-stimulated insulin production. The apoptosis was analyzed by flow cytometry. Thiaoollyl blue viability assay was applied to exam the number of viable cells.


RESULTS
The recombined adenovirus-rIGF-1 was constructed successfully and its titer was about 4.0x10(8) pfu/ml. The rIGF-1 was expressed in the RINm5F cells and cell culture supernatants. rIGF-1 expression could inhibit islet cells apoptosis, significantly decrease the level of NO induced by streptozotocin and significantly increase insulin secretion and cells viability.


CONCLUSIONS
These results suggested that the high concentration of rIGF-1 in cultured islets may be beneficial in maintaining islet beta cells function and protecting islet beta cells from apoptosis-mediated factors. The apoptosis induced by STZ may be NO-dependent.
Chen
Zhi-hong
ZH
Department of Peadiatrics, Shandong University Qilu Hospital, Jinan 250012, China.
Li
Tang
T
Luo
Bing
B
Zhu
Wei
W
Sun
Ruo-peng
RP
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003920:Diabetes Mellitus / Q000235:genetics / Q000628:therapy; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D013311:Streptozocin / Q000494:pharmacology*
D007328:Insulin; D013311:Streptozocin; D007334:Insulin-Like Growth Factor I