化浊清脂方对2型糖尿病合并高脂血症大鼠内脂素水平、氧化应激及肝细胞凋亡的影响

Effect of Prescription of Turbidity-Resolving and Lipid-Eliminating on Visfatin Level,Oxidative Stress and Hepatocyte Apoptosis in Rats with Type 2 Diabetes Complicated with Hyperlipidemia

目的:探讨化浊清脂方对2型糖尿病合并高脂血症大鼠内脂素水平、氧化应激及肝细胞凋亡的影响。方法:选取健康SD雄性大鼠60只,喂养1周后,根据体重随机区组法将6只纳入正常组,54只纳入模型组。正常组大鼠继续采用常规饲料喂养,模型组大鼠采用高糖高脂饲料喂养。8周后,模型组大鼠腹腔注射2%链脲佐霉素制备2型糖尿病合并高脂血症大鼠模型,选取与正常组存在显著差异的36只大鼠纳入研究,通过随机区组法将36只大鼠分为模型组、二甲双胍组、脂必妥胶囊组及化浊清脂方(低、中及高剂量)组,每组6只。对各组大鼠进行灌胃给药治疗,治疗1个月后,对比各组大鼠内脂素水平、氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)、晚期糖基化终末产物(AGEs)及晚期蛋白质氧化产物(AOPP)]、肝细胞凋亡及B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)表达情况。结果:模型组大鼠内脂素水平高于其他组,且与正常组和化浊清脂方低、中及高剂量组相比的差异均有统计学意义(P<0.05);模型组大鼠SOD水平低于其他组,AGEs、MDA及AOPP水平高于其他组,上述氧化应激指标与正常组和化浊清脂方中、高剂量组相比的差异均有统计学意义(P<0.05),AOPP水平与化浊清脂方低剂量组的差异有统计学意义(P<0.05)。模型组大鼠的细胞凋亡程度高于其他组,与正常组、二甲双胍组及化浊清脂方低、中剂量组相比的差异均有统计学意义(P<0.05)。模型组大鼠静止期(G0/G1期)细胞占比明显高于正常组,差异有统计学意义(P<0.05);略高于各给药组,但差异均无统计学意义(P>0.05)。模型组大鼠的增殖指数明显低于正常组,差异有统计学意义(P<0.05);略低于各给药组,但差异均无统计学意义(P>0.05)。模型组大鼠的Bax蛋白表达免疫荧光指数(FI)值明显高于正常组、化浊清脂方低、中及高剂量组;Bcl-2蛋白表达FI值明显低于正常组、二甲双胍组及化浊清脂方低、中剂量组;Bax/Bcl-2蛋白表达水平明显高于正常组、二甲双胍组和化浊清脂方低、中及高剂量组,差异均有统计学意义(P<0.05)。结论:化浊清脂方可以有效降低2型糖尿病合并高脂血症大鼠内脂素水平,改善大鼠氧化应激状态;并通过增强Bcl-2蛋白表达、减弱Bax蛋白表达来降低大鼠肝细胞凋亡率。

OBJECTIVE:To probe into the effect of prescription of turbidity-resolving and lipid-eliminating on visfatin level,oxidative stress and hepatocyte apoptosis in rats with type 2 diabetes complicated with hyperlipidemia.METHODS:60 healthy SD rats were selected,after 1-week feeding,6 rats were brought into normal group and 54 rats were brought into model group according to random weight grouping method.The normal group was fed with conventional fodder,while the model group was fed with high-sugar and high-fat fodder.After 8 weeks,rats in the model group were intraperitoneally injected with 2%streptozotocin to prepare rat model of type 2 diabetes complicated with hyperlipidemia,36 rats with significant differences from the normal group were selected for study and divided into model group,metformin group,Zhibituo capsule group and turbidity-resolving and lipid-eliminating group(low,medium and high dose)via randomized block,with 6 rats in each group.Intragastric administration was given to rats in each group,after 1-month treatment,the visfatin level,oxidative stress[superoxide dismutase(SOD),malonaldehyde(MDA),advanced glycosylation end products(AGEs)and advanced oxidation protein products(AOPP)],hepatocyte apoptosis,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)expression were compared among each group.RESULTS:The visfatin level in model group was significantly higher than other groups,which was statistically different with that of normal group,low,medium and high dose of turbidity-resolving and lipid-eliminating group(P<0.05);the SOD level of model group was significantly lower than other groups,while the AGEs,MDA and AOPP levels were significantly higher than other groups,which were statistically different with those of normal group,medium and high dose of turbidity-resolving and lipid-eliminating group(P<0.05),the AOPP level of model group was statistically different with that of low dose of turbidity-resolving and lipid-eliminating group(P<0.05).The cell apoptosis of model group was significantly higher than other group,which was statistically different with that of normal group,metformin group,low and medium dose of turbidity-resolving and lipid-eliminating group(P<0.05).Cell proportion of model group at stationary phase(G0/G1)was significantly higher than that of normal group,with statistically significant difference(P<0.05);which was slightly higher than each administration group,with no statistically significant difference(P>0.05).The proliferation index of model group was significantly lower than that of normal group,with statistically significant difference(P<0.05);which was slightly lower than each administration group,with no statistically significant difference(P>0.05).The fluorescence index(FI)of Bax protein expression of model group was significantly higher than that of normal group,low,medium,high dose of turbidity-resolving and lipid-eliminating group;the FI of Bcl-2 protein expression was significantly lower than that of normal group,metformin group,low and medium dose of turbidity-resolving and lipid-eliminating group;the expression level of Bax/Bcl-2 protein were significantly higher than that of normal group,metformin group,low,medium and high dose of turbidity-resolving and lipid-eliminating group,with statistically significant differences(P<0.05).CONCLUSIONS:Prescription of turbidity-resolving and lipid-eliminating can effectively reduce the visfatin level in rats with type 2 diabetes complicated with hyperlipidemia,and improve oxidative stress;which can reduce hepatocyte apoptosis by enhancing the expression of Bcl-2 protein and weakening the expression of Bax protein.