Experimental Study On The Effect Of Qingzhi Tongmai Serum On Apoptosis Related Factors Of HUVECs Induced By H ₂ 2

Purpose：Research of Qingzhi Tongmai serum on changes of HUVECs apoptosisrelated factor caspase-3and Bcl-2, the expression of Bax induced by H2O2, toexplore the mechanism of Qingzhi Tongmai Recipe in preventing AS from genelevel, and provide a theoretical basis for the prevention and treatment of ASwith traditional Chinese Medicine.Material and method：1. The purchase of32New Zealand white rabbits, were randomly divided into4groups, namely control group, Qingzhi Tongmai low dose group, middle dose groupof Qingzhi Tongmai, Qingzhi Tongmai high dose group,8rats in each group, halfmale and half female. Control group was fed normalally, Qingzhi Tongmai groupswere intragastric administration of low dose (10%,3g crude drug/kg/d), mediumdose (33.3%,10g crude drug/kg/d), high dose (100%,30g crude drug/kg/d)Decoction of Chinese medicine. Each group according to15ml/kg/times, to beadministered,2times daily administration, sooner or later each time,continuous7d, heart blood last gavage,2h, serum was separated by centrifugation,combined with serum.2. By using the method of cell culture in vitro, putting the HUVECs culturedin DMEM (10%fetal bovine serum+100g/ml+100IU/ml streptomycin, penicillin)in37℃,5%CO2sterile constant temperature incubator culture. When the cellsreached80%together, by trypsin digestion, cultured. The cultured cells weredivided into5groups: control group: in the solution with26h continuous cultureblank serum DMEM culture; model group: Cultured in DMEM continuous24h culturefluid, added with100μ mol/L H2O22h, copy the oxidative cell injury model; Qingzhi Tongmai low dose group: Cultured in DMEM add10%liquid Qingzhi Tongmaiserum continuous culture after24h, add H2O22h; clear grease through dose grouppulse: Cultured in DMEM with33.3%liquid Qingzhi Tongmai serum continuousculture after24h, add H2O22h; Qingzhi Tongmai high dose group: Cultured in DMEMwith100%liquid Qingzhi Tongmai serum continuous culture after24h, add H2O22h.3. Using RT-PCR and Western blot methods for detection of caspase-3and Bcl-2,Bax mRNA and protein expression level changes.Results：1. RT-PCR detection showed: The blank group caspase-3and Bcl-2, Bax mRNA wasexpressed at low levels,compared with the blank group, model group, mRNAsignificantly reduced the expression of Bcl-2, caspase-3, Bax mRNA expressionwas significantly increased (P<0.01); compared with the model group, the fatTongmai low, middle and high dose group, Bcl-2mRNA expression with the increaseof drug concentration gradually increased, caspase-3, Bax mRNA expression withthe increase of drug concentration and gradually decreased, in a dose-dependentmanner, the middle dose group is more obvious, the most significant of high dosegroup (P<0.01).2. Western blot detection showed: The blank group caspase-3and Bcl-2, Baxprotein was expressed at low levels,compared with the control group, the Bcl-2expression in model group was significantly decreased, the expression ofcaspase-3, Bax protein were significantly increased (P<0.01); compared with themodel group, the fat Tongmai low, middle and high dose group, the expressionof Bcl-2protein increased gradually with the concentration increased, theexpression of caspase-3, Bax protein with drugs concentration decreased, in adose-dependent manner, the middle dose group is more obvious, the mostsignificant of high dose group (P<0.01).Conclusion：1. H2O2enables the Bcl-2mRNA and protein expression of HUVECs decreased si gnificantly, caspase-3, Bax mRNA and protein expression was up-regulated.2. Qingzhi Tongmai serum can make Bcl-2mRNA and protein expression decreasedthe expression of caspase-3, Bax, mRNA and protein.3. Qingzhi Tongmai Recipe could inhibit apoptosis through regulating the expression of apoptosis related factor caspase-3and Bcl-2, Bax, so as to controlthe action of AS.