朱日很滴丸对动脉粥样硬化模型兔MMP-9表达量的影响

Effect of Zhurihen Dropping Pills on the expression of MMP-9 in atherosclerosis model rabbits

目的研究朱日很滴丸对动脉粥样硬化模型兔基质金属蛋白酶9(MMP-9)表达量的影响。方法1.采用高脂饲料+牛血清白蛋白(250 mg·kg-1)对日本大耳白兔进行造模,第8周造模成功后,朱日很滴丸低、中和高剂量组分别灌胃朱日很滴丸溶液116,367和1160 mg·kg-1,普罗布考组灌胃普罗布考溶液26 mg·kg-1,每日1次,连续4周。实验期间,溶媒对照组给予普通饲料,其余组结予高脂饲料。实验结束后采集各组兔血清及主动脉,通过Elisa法和免疫组化法(IHC-P)测定朱日很滴丸对动脉粥样硬化模型兔血清和主动脉MMP-9表达量的影响;2.将氧化性低密度脂蛋白(ox-LDL)与人脐静脉内皮细胞(HUVEC)共同孵育,建立内皮细胞损伤模型,加入朱日很滴丸含药血清进行干预。通过实时荧光定量PCR法(qPCR)和蛋白免疫印迹法(Western Blot)检测朱日很滴丸含药血清对内皮细胞氧化损伤模型中MMP-9的mRNA水平和蛋白含量的影响。结果与模型组相比,朱日很滴丸低、中和高剂量组兔血清与主动脉斑块中MMP-9表达量均显著下降,经朱日很滴丸低、中和高剂量含药血清处理的细胞中MMP-9的mRNA水平和蛋白含量均显著下降。结论朱日很滴丸可降低动脉粥样硬化模型兔MMP-9的表达量,具有临床应用价值。

Objective To study the effect of Zhurihen Dropping Pills on the expression of matrix metalloproteinase 9(MMP-9)in atherosclerosis model rabbits.Methods 1.Japanese rabbits were fed with high-fat diet plus bovine serum albumin(250 mg·kg-1).After the successful establishment of the model at the 8th week,the low,middle and high dose groups were given Zhurizhu Dropping Pills solution by gavage,the doses were respectively 116,367 and 1160 mg·kg-1.Meanwhile the probucol group was given probucol solution once a day for 4 weeks by gavage,the dose was 26 mg·kg-1.During the experiment,the solvent control group was given ordinary diet and the rest groups were given high fat diet.After the experiment,the serum and aorta of rabbits were collected,and the effects of Zhurihen Dropping Pills on the content of MMP-9 in the serum and aorta of atherosclerosis model rabbits were determined by Elisa and immunohistochemistry(IHC-P).2.The oxidized low density lipoprotein(ox-LDL)was incubated with human umbilical vein endothelial cells(HUVEC)to establish the injury model of endothelial cells.The drug-containing serum of Zhurihen Dropping Pills was added for intervention.Real-time fluorescence quantitative PCR(qPCR)and Western Blot were used to detect the effects of Zhurihen Dropping Pills on the mRNA and protein levels of MMP-9 in endothelial cell oxidative damage model.Results Compared with the model group,the levels of MMP-9 in serum and aortic plaque of rabbits in low,middle and high dose groups of Zhurihen Dropping Pills were significantly decreased;the mRNA and protein levels of MMP-9 in serum cells treated with low,middle and high dose drug containing serum were significantly decreased.Conclusion Zhurihen Dropping Pills can reduce the expression of MMP-9 in atherosclerosis model rabbits and have clinical application value.