蚓激酶对转化生长因子-β1诱导的人肾小管上皮细胞Src激酶及黏着斑激酶表达的影响  

Effect of lumbrokinase on the synthesis of Src and FAK induced by TGF-β1 in human renal tubular epithelial cells

作　　者：魏晓露 李月红 李国霞 Wei Xiaolu;Li Yuehong;Li Guoxia(Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;Department of Nephrology, Tianjin Nankai Hospital, Tianjin 300100, China;International Medical School, Tianjin Medical University, Tianjin 300070, China)

机构地区：[1]天津中医药大学,301617 [2]天津南开医院肾内科,300100 [3]天津医科大学国际医学院,300070

出　　处：《国际中医中药杂志》2019年第8期847-851,共5页International Journal of Traditional Chinese Medicine

基　　金：天津医科大学留学生科技创新项目(2110-2GJ003).

摘　　要：目的观察蚓激酶对TGF-β1诱导人肾小管上皮细胞a-平滑肌肌动蛋白(a-smooth muscle actin, a-SMA)、纤维连接蛋白(fibronectin, FN)、黏着斑激酶(focal adhesion kinase, FAK)和Src家族蛋白酪氨酸激酶表达的影响。方法将人肾小管上皮细胞按随机数字表法分为正常组,模型组,贝那普利组及蚓激酶低、中、高剂量组。除正常组外,其余各组加入10 μg/ml的TGF-b1溶液进行干预,30 min后,贝那普利组加入盐酸贝那普利10 μmol/L进行干预,蚓激酶低、中、高剂量组分别加入蚓激酶30、60、120 U/ml进行干预。培养24 h后,采用Western blot法和荧光定量PCR检测a-SMA、FN、FAK和Src激酶蛋白及基因表达。结果与模型组比较,蚓激酶低、中、高剂量组a-SMA[(0.84±0.14)、(0.72±0.08)、(0.69±0.05)比(1.24±0.03)]、FN[(0.59±0.09)、(0.55±0.11)、(0.44±0.08)比(0.83±0.18)]、FAK[(0.94±0.04)、(0.79±0.05)、(0.70±0.02)比(1.29±0.07)]、Src激酶[(0.87±0.20)、(0.78±0.15)、(0.71±0.11)比(1.23±0.01)]蛋白表达降低(P<0.05),a-SMA mRNA[(3.13±0.62)、(2.76±0.14)、(2.15±0.33)比(4.12±0.32)]、FN mRNA[(3.08±0.34)、(2.78±0.17)、(2.49±0.11)比(4.34±0.06)]、FAK mRNA [(1.73±0.23)、(1.63±0.36)、(1.57±0.27)比(2.61±0.59)]、Src激酶mRNA[(2.11±0.17)、(1.78±0.25)、(1.71±0.22)比(2.78±0.47)]表达降低(P<0.05)。结论蚓激酶可通过调节TGF-b1诱导的人肾小管上皮细胞FAK、Src激酶的表达,减少a-SMA、FN生成,从而阻抑肾纤维化的发展。Objective To observe the regulation of lumbrukinase on the expression of a-smooth muscle actin, fibronectin, focal adhesion kinase and Src kinase induced by transforming growth factor b1 in human renal tubular epithelial cells. Methods According to random number table method, the human renal tubular epithelial cells were divided into the normal group, TGF-b1 model group, benazepril group, lumbrukinase low, medium and high dose group. Except the normal group, the other groups were treated with TGF-b1 10 μg/ml. After 30 minutes, the benazepril group added benazepril 10 μmol/L, and the low, medium, high dose groups of lumbrukinase were respectively treated with lumbrokinase 30, 60, 120 U/ml to intervene. After 24 hours of cultivation. Western blotting and real-time PCR were used to detect the expression of a-SMA, FN, FAK and Src. Results Compared with the model group, the expressions of a-SMA (0.84 ± 0.14, 0.72 ± 0.08, 0.69 ± 0.05 vs. 1.24 ± 0.03) and FN (0.59 ± 0.09, 0.55 ± 0.11, 0.44 ± 0.08 vs. 0.83 ± 0.18) and FAK (0.94 ± 0.04, 0.79 ± 0.05, 0.70 ± 0.02 vs. 1.29 ± 0.07) and Src (0.87 ± 0.20, 0.78 ± 0.15, 0.71 ± 0.11 vs. 1.23 ± 0.01) proteins in the high doses of lumbrical kinase group were significantly lower than those in the model group (P<0.05), the expressions of a-SMA (3.13 ± 0.62, 2.76 ± 0.14, 2.15 ± 0.33 vs. 4.12 ± 0.32) and FN (3.08 ± 0.34, 2.78 ± 0.17, 2.49 ± 0.11 vs. 4.34 ± 0.06) and FAK (1.73 ± 0.23, 1.63 ± 0.36, 1.57 ± 0.27 vs. 2.61 ± 0.59) and Src (2.11 ± 0.17, 1.78 ± 0.25, 1.71 ± 0.22 vs. 2.78 ± 0.47) mRNA in the high doses of lumbrical kinase group decreased significantly (P<0.05). Conclusions Lumbrokinase may prevent the development of renal fibrosis by regulating the expression of FN, FAK and and reducing the production of a-SMA, FN.

关 键 词：纤维化 蚓激酶 A-平滑肌肌动蛋白 src-族激酶类 上皮-间质转分化 上皮细胞 

分 类 号：R96[医药卫生—药理学]