The Effect Of Insulin On Albumin MRNA Expression In Rat Hepatocytes With High Glucose In Vitro

Objective To observe the effect of different dose insulin on albumin mRNA expression in co-culture Kupffer cells with hepatocyte in vitro. Methods In-situⅣcollagenase two-step perfusion method was used to digest rat liver, low speed centrifugalization was applied to isolate parenchymal hepatic cells and density gradient centrifugtion by percoll fluid to isolate Kupffer cells. Hepatocyte cells were inoculated into culture plate to co-culture with Kupffer cells in the proportion six to one; the cells were incubated in mediums containing 11.1mmol/L glucose and different dose insulin[insulin(IU):glucose(g)=2:1,Group 1;1:1,Group 2;1:2,Group 3;1:4,Group 4] were used to manage the co-culture cells experiment model. The albumin mRNA of hepatocytes was extracted and assessed by reversetranscription-polymerase chain reaction(RT-PCR) and the level of albumin,IL-1,IL-6,TNF-αin the supernatant were measured by ELISA at 4h,24h after intervention respectively. Results At 4h,24h, the level of IL-1,IL-6,TNF-αin the culture supernatant fluid in the high concentration of glucose (11.1mmol/L) was increased significantly and the level of albumin and expression of albumin mRNA was decreased significantly along with reduction of insulin dosage. Conclusion Hepatocytes co-culture with Kupffer cells can be carried out under eligible culture condition, the secretive function of Kupffer cells and hepatocytes are maintained.In the circumstance of high concentration glucose (11.1mmol/L), insulin may play an important role in anti-inflammatory response induced by inhibitting cytokines generation and exert promoting effects on transcription of rat hepatocyte albumin mRNA and albumin synthesization of rat hepatocyte.