Effect On Apoptosis By SMS RNA Interfence In Smmc-7721Cells(the Plasma Level Of SM And CETP As Well As PLTP Impact On The Generate Effect Of Ⅱ Type Diabetes Mellitus)

Recently reported that it is an affinity between Ⅱ type diabetes mellitus and atherosclerosis，butinflammation maybe considered a common pathological mechanism. As we know，Sphingomyelin is anindependent risk factor for atherosclerosis，therefore we infer Sphingomyelin maybe related to2typediabetes mellitus. More, Ceramide is considered as regulating signal factory in cell apoptosis andproliferation.Furthmore, Sphingomyelin and Ceramide are important chemical compround forSphingolipid metabolism meanwhile Sphingomyelin synthase is the key enzyme which catalytics substratephosphatidylcholine and ceramide generation product of sphingomyelin and diacylglycerol.Atherosclerosisand2type diabetes mellitus exist the disorders of lipid metabolism. So, We will take SMMC-7721cell asexperimental model and observe the apoptosis rate of SMMC-7721cell after SMS RNA interfuencemeanwhile we research the relationship between atherosclerosis and diabetes mellitus through measuringsphingomyelin and phospholipid transfer protein as well as cholesterol ester transfer protein in plasmafor Ⅱ type diabetes mellitus.Objective: To observe the cell apoptosis rate and the activities by SMS RNA interfuence as well asdiscuss the relationship between atherosclerosis and diabetes mellitus，for providing more experimentaldata for the pathogenesis and clinical treatment for diabetes mellitus.Methods: Take SMMC-7721cell as experimental model, inducing cell apoptosis and by MTTscreening the dose for TNF-α, making SMS gene silence by RNA interference technology, detecting theenzyme activities of SMS after RNA interfuence by TLC, FITC-annexin V/PI flow cytometry and Hoechst33258/PI were used to detect the apoptotic cells, Western blotting analysis was used to assay theexpression of apoptosis-related proteins. Diagnosis of Ⅱ type diabetes mellitus by according to2009American Diabetes Association. The level of SM and CETP as well as TLTP of diabetic blood serum weredetected by enzyme mearsuring and fluorophotometric spectrometer assay.Results: MTT results have shown that TNF-α has the potential apoptosis effects on theSMMC-7721cells at24h as well as has a dose-and time-dependent manner. Take the concentration ofTNF-α as200ng·μL-1for experimental study.It was found that the activities of SMS were decreased and cell apoptosis rate was higher by FITC-annexin V/PI flow cytometry and Hoechst33258/PI assay afterSMS RNA interfuence,compared with the control group（P<0.05）.Expression of apoptosis relatedproteins caspase-8and caspase-3increased significantly compared with the control group. The bloodserum results indicated the level of SM、CETP and TLTP were higher than the control group （P<0.05）.Conclusions:1.The enzyme activities of SMS decreased meanwhile the cell apoptosis rateincreased after SMS RNA interfuence.2. The increased levels of SM and CETP as well as PLTP promotethe generation for Ⅱ type diabetes mellitus patients.