The Effects Of L-carnitine On The Adiponectin Receptors Expression Of RMCs Induced By Intermittent High Glucose

Background & objectiveThe project intended to study the effect of intermittent high glucose on RMCs adipone ctin receptors (AdipoR) expression, and to explore the change after suppling L-CN. To ide ntify the important role L-CN plays in.If we earn what we want , then we can push the L-ca rnitine in improving myocardial energy metabolism in diabetes at the same time, by increa sing the expression of adiponectin receptors to reduce insulin resistance and diabetes relate d obesity. And through the subsequent animal experiments, clinical observations verify this idea. L-carnitine can bring the Gospel for people suffer from ischemic heart disease and di abetes.Research MethodsFirst, the myocardial cells (RMCs) Culture and identificationSD rat cardiac cell line (purchased from Shanghai Institute), cell recovery, DMEM me dium (6.1mmol / L), normal culture,observe cell morphology, trypan blue staining and iden tification of cell survival. Second, intermittent high glucose on RMCsAfter the first experiment with cells serum-free DMEM (6.1mmol / L) pretreatment m edium 12h, it is in synchronization state. Experimental groups: normal culture group, hig h glucose medium group, the intermittent high glucose culture medium cell group, L-CN tre atment groups. The centration of intermittent high glucose (rotation per 8h adding 5.5mmol / L or 20mmol / L glucose solution) were cultured routinely 5d. Detected by CCK8. Third, L-CN stimulation RMCs pretreatment by intermittent high glucose. (A) different concentrations of L-CN stimulus 24h.L-CN treatment group (working concentrations 102μΜ, 2.5×102μΜ, 5×102μΜ, 103μΜ, 104μΜ). (B) a certain concentration of L-CN stimulus at different timeL-CN working concentration 5×102μΜ, L-CN treatment (5×102μΜstimulate 0h, 4h, 8h, 12h, 24h, 36h, 48h), Fourth, detectionCCK8 detection RMCs proliferation activation. Real time PCR detection AdipoR mR NA expression changes. Fifth,Statistical analysis Data with x±s represent,all statistical data are handled by SPSS l8.0 software,comparison between groups using single factor analysis of variance,the two groups using t test, homogeneity of variance compared with the LSD method; Heterogeneity of variance with Dunnett's Methods. P <0.05 considered statistically significant.ResultsFirst,identification of cell counts and survivalTrypan blue staining RMCs more than 95% survival rate,meet the experimental needs. Second,CCK8 detecte RMCs proliferation activation.(A) CCK8 detecte activation of proliferation with different concentrations of L-CN stim ulite after 24h.Compared with ordinary culture medium, high glucose group OD value decresed but not obvious; Myocardial cells in intermittent high glucose group OD values decresed, compare d with the normal medium was significant difference (P <0.05); L-CN (5×102μΜ) treatm ent group compared with the intermittent high glucose,the OD values incr eased was statis tically significant (P <0.05); L-CN (2.5×102,103,104μΜ) treated compa red with the inter mittent high glucose increase in OD values,but not statistically significant (P> 0.05); L-CN (105,106μΜ) treated compared with the intermittent high glucose in OD values decreased.(B) CCK8 detecte activation of proliferation after a certain concentration of L-CN stimu lite different time.Compared with ordinary culture medium, high glucose group OD value decreased but n ot obvious,the OD value of the different intermittent high glucose group was significant de crease (P <0.05),L-CN (5×102μΜstimulation 4h , 8h, 12h) groups compared with interm ittent high glucose group OD values increased, but not statistically significant,L-CN (5×102μΜstimulate 24h, 36h, 48h) group compared with intermittent high glucose group OD values increased (P <0.05).Third, adipoR mRNA expression after stimulating by different concentrations of L-CN 24 hCompared with ordinary culture medium, the intermittent high glucose treatment Adipo R mRNA expression was significantly decreased (P <0.0 1),and the intermittent high gluc ose group treated by adding L-CN groups, AdipoR1,AdipoR2 mRNA expression were sign ificantly higher (P <0.01),and when L-CN working concentration of 5×102μΜAdipoR m RNA increased most significantly (between the two P <0.01).Fourth,adipoR mRNA expression when a certain concentration of L-CN stimulate for diff the L-CN (5×102μΜ) treatment of myocardial cells in each phase, AdipoR1 mRNA ex pression were increased, and when the drug increased significantly at 24h (P <0.05 .) Adip oR2 mRNA compared to Adipo R1 performance characteristics of the instability. And whe n dealing 4h AdipoR2 mRNA expression did not rise but fall,12h increased but not statisti cally significant (P> 0.05 ),the increase of AdipoR2 mRNA 24h, the most significant (P <0. 01).ConclusionThe intermittent high glucose group compared to simple high glucose stronger effect on cell damage, L-CN can reduce to some extent,and the effective concentration of drug betw een the 102-104μΜ.L-CN showed that the role of promoting value-added time-dependent c haracteristics.L-CN treated RMCs AdipoR1 mRNA, AdipoR2 mRNA were significantly higher (P <0. 01), when L-CN working concentration of 5×102μΜ, AdipoRmRNA expression increase ed more significantly. L-CN RMCs AdipoR1 mRNA, AdipoR2 mRNA showed significan tly increased time-dependent, after L-CN (5×102μΜ) treated for 24 hours the AdipoR mR NA increased most significantly (P <0.0 5).