糖通饮含药血清对高糖培养HBZY-1细胞microRNA-21、Smad7以及α-SMA表达的影响  

Effect of tangtongyin drug-containing serum on the expression microRNA-21,Smad7,andα-SMA in high-sugar cultured HBZY-1 cells

作　　者：伏红颖 潘艳伶 陈俞如 陈洪民 FU Hongying;PAN Yanling;CHEN Yuru;CHEN Hongming(Teaching and Research Department of Traditional Chinese Medicine,School of Clinical Medicine,Guizhou Medical University,Guiyang 550000,China;Department of Traditional Chinese Medicine,the Affiliated Hospital of Guizhou Medical University,Guiyang 550000,Guizhou,China)

机构地区：[1]贵州医科大学临床医学院中医学教研室,贵州贵阳550000 [2]贵州医科大学 [3]贵州医科大学附属医院中医科,贵州贵阳550000

出　　处：《贵州医科大学学报》2022年第6期628-634,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82060837);贵州医科大学附属医院2020年国家自然科学基金培育项目(gy5ynsfc〔2020〕-3)。

摘　　要：目的观察临床经验方糖通饮含药血清对高糖环境下大鼠肾小球系膜细胞(HBZY-1)microRNA-21(miR-21)、Smad7以及α-平滑肌蛋白(α-SMA)表达的影响。方法30只SPF级雄性SD大鼠随机分为空白血清组和糖通饮含药血清组,糖通饮含药血清组大鼠予以糖通饮[12.4 g/(kg·d)]灌胃,空白血清组大鼠予以等体积的双蒸水灌胃,连续灌胃1周后股动脉取血获取糖通饮含药血清;将HBZY-1细胞分为正常糖组、高糖组、高糖+低、中、高浓度糖通饮含药血清组(糖通饮含药血清浓度分别为5%、10%、15%),CCK-8法检测各组细胞增殖率,根据细胞增殖被抑制情况筛选出最佳浓度糖通饮含药血清作为后续实验的处理因素;将HBZY-1细胞分为正常糖+空白血清组、高糖+空白血清组、高糖+最佳浓度糖通饮含药血清组,CCK-8法检测各组细胞增殖情况,实时荧光定量PCR法检测各组细胞miR-21表达以及Smad7和α-SMA mRNA表达,Western blot法检测各组细胞Smad7、α-SMA蛋白的表达。结果与高糖组比较,低、中、高浓度糖通饮含药血清组细胞增殖率均降低(P<0.05),但中、高浓度糖通饮含药血清组的细胞增殖率比较,差异无统计学意义(P>0.05),因此后续实验选用中浓度作为最佳浓度糖通饮含药血清;对高糖环境下的HBZY-1细胞予以最佳浓度糖通饮含药血清干预后结果显示,与高糖+空白血清组相比,高糖+最佳浓度糖通饮含药血清组细胞增殖被抑制,HBZY-1细胞中miR-21表达降低,α-SMA的mRNA和蛋白表达降低,Smad7的mRNA和蛋白表达升高(P<0.05)。结论糖通饮可能通过调控miR-21、Smad7和α-SMA表达抑制高糖环境中HBZY-1细胞的增殖。Objective To investigate the effect of tangtongyin drug-containing serum on the expression of microRNA-21(miR-21),Smad7,andα-smooth muscle actin(α-SMA)in rat glomerular Mesangial cells(HBZY-1)in high glucose environment.Methods A total of 30 SPF male SD rats was randomly divided into blank serum group and tangtongyin drug-containing serum group.Rats in tangtongyin drug-containing serum group were intragastric administred with 12.4 g/(kg·d)tangtongyin,and rats in blank serum group were fed with the same volume of double distilled water.After continuous intragastric administration for 1 week,blood was drawn from the femoral artery for tangtongyin drug-containing serum.HBZY-1 cells were divided into normal glucose group,high glucose group,high glucose+low/medium/high concentration tangtongyin drug-containing serum group(concentration was respectively 5%,10%,15%).The cell proliferation rate of each group was detected by CCK-8 assay,and the best concentration of tangtongyin drug-containing serum was selected according to the inhibition of cell proliferation for treatment factor of follow-up experiment.The cells were divided into:normal glucose+blank serum group,high glucose+blank serum group,high glucose+optimal concentration tangtongyin drug-containing serum group.The cell proliferation was detected by CCK-8 assay,the expression of miR-21,Smad 7,andα-SMA mRNA were detected by real-time fluorescence quantitative PCR,and the protein expression of Smad7 andα-SMA were detected by Western blot.Results Compared with high glucose group,the cell proliferation rate in the low,medium and high concentration tangtongyin drug-containing serum groups decreased(P<0.05),but there was no significant differences between the medium and high concentration tangtongyin drug-containing serum groups(P>0.05).Therefore,the medium concentration was selected as the optimal concentration of tangtongyin drug-containing serum in the follow-up experiment.After the intervention of the optimal concentration of tangtongyin drug-containing serum o

关 键 词：肾小球系膜细胞 MICRORNA-21 SMAD7 α-平滑肌蛋白 大鼠 SD 糖通饮含药血清 高糖环境 

分 类 号：R285.5[医药卫生—中药学]