The Effect On Cell Proliferation And Apoptosis Of Vascular Smooth Muscle Cell By Hypoxia/reoxygen And The Empirical Study Of Scorpion Venom's Intervention

In a certain time to recovery the blood supply to the cardiac muscle whichblood supply interrupted by a short time, can induce more severe injury in theoriginal ischemic cardiac muscle than before, called ischemia/reperfusion injury(IRI). Including cellular adhension molecule (CAM),heat shock protein (HSP),nuclear factor (NF),nitrogen monoxidum (NO),apoptosis and so on , multiplemechanism participates in IRI. PURPOSE: In this experiment we adopted primary cell culture of SMC, toobserve the change of cell proliferation and apoptosis of SMC afterhypoxia/reoxygen and give scorpion venom to the cells on the H/R base IRI, aswell as the change of expression of apoptosis related gene bcl-2,bax , to approachthe effect of hypoxia/reoxygen and scorpion venom on VSMC. METHOD: SMC be cultured by tissue-sticking method in the experiment, weuse 4-6 generated cells. Cells were random divided to: control,H/R model group,model +SV groups, among the model+SV groups, SV divided to 5,10,20,40mg/L different density. Use simulated hypoxia liquid and simulated reoxygenliquid to culture SMC, set up hypoxia/reoxygen (H/R) model. To determine thecell proliferation by MTT, to decect the apoptosis by TUNEL, to evaluate theexpression of bcl-2,bax by immunohistochemical method. The experimental datawas denoted to mean ±standard deviation(X±S), use sample mean t-checkoutto statistics analyze the numerus compare of each group. RESULT: Experimental result shows that: (1)to compare the result of eachgroup's cell-proliferation, normal control A value is 1.70±0.24, after H/R, H/Rmodel A value is 2.23±0.27, there is a significant difference between the twogroups in statistics (P<0.01, n=8). Postischemical SMC cell proliferation isgrowed in number;(2)On H/R injury model ground, we give different density SV,5mg/L SV group's A value is 1.99±0.28, 20mg/L SV group's A value is 2.13±0.24, compared with normal control, there is statistical significance (P<0.05,n=8);10mg/L SV group's A value is 1.92±0.16, 40mg/L SV group's A value is 1.89±0.74, compared with H/R model group, there is statistical significance(P<0.05,n=8). The result hints that 10mg/L,40mg/L SV has a inhibit effect onSMC cell proliferation by IRI;(3)to compare apoptosis rate of normal control withthat of H/R model, normal control is 26.4±1.4%, and H/R model group is17.4±0.9%, there is statistical significance (P<0.05,n=8). Hint that SMC apoptosis isreduced after H/R;(4)5mg/L SV group apoptosis ratio is 25.6±2.8%, 10mg/L SVgroup apoptosis ratio is 33.5±3.5%, 40mg/L SV group apoptosis ratio is 34.5±5.2%, respectively compared with normal control and H/R model group, each hasstatistical significance (P<0.05,n=8);20mg/L SV group apoptosis ratio is 20.1±2.4%, compared with normal control, there is statistical significance (P<0.05,n=8).The result shows that 5mg/L,10mg/L,40mg/L SV can promote apoptosis onSMC after H/R injury;(5)Compared with normal control, bcl-2,bax proteinmasculine rate and bcl-2/bax ratio in H/R model group are significant changed(P<0.05,n=8);5mg/L,10mg/L,20mg/L,40mg/L SV group's bcl-2/bax ratioheightened than H/R model group, there is statistical significance (P<0.05). Theresult shows that SV may influence apoptosis through bcl-2/bax pathway.CONCLUSION:1,Hypoxia/reoxygen can increase VSMC proliferation and induce VSMCapoptosis.2,Scorpion venom has an inhibiting effect of cell proliferation of VSMC andcan promote apoptosis of VSMC after hypoxia/reoxygen.3,Hypoxia/reoxygen or SV may through bcl-2/bax pathway to mediateapoptosis.Sum up, SV can inhibit cell proliferation and promote apoptosis of VSMCafter hypoxia/reoxygen, and its possible mechanism is through related gene bcl-2,bax pathway, cause bcl-2/bax ratio raise up to mediate apoptosis.New ideas points:1,The first time to establish hypoxia/reoxygen (H/R) model on vascularsmooth muscle cell, observe the change of VSMC cell proliferation and apoptosisafter hypoxia/reoxygen.2,Supply scorpion venom (SV) to VSMC based on H/R model, to observeSV's influence on cell proliferation and apoptosis.3,The experiment confirm that bcl-2/bax way mediates SMC apoptosis afterH/R injury and SV intervention.RESEARCH SIGNIFICANCE:To prevent and cure ischemical reperfusion injury is a hot spot incardiovascular research domain, our experiment established hypoxia/reoxygeninjury model on SMC first time, observe ischemia reperfusion injury on SMC.Scorpion venom is our country traditionary Chinese medicine, which has a lot ofbiological activities, such as analgesia,antitumor,anti-epileptic,fibrolysic systemand ion channel and so on. Our experiment study conduce to illuminatehypoxia/reoxygen injury effect on SMC, at the same time prove SV's protectionof SMC after H/R injury, offer new experimental and theoretical evidence tofurther develop SV to applicate anti-ischemical reperfusion injury...