水飞蓟宾对高糖作用下大鼠腹膜间皮细胞整合素连接激酶、转化生长因子β1及α平滑肌肌动蛋白表达的影响  

Effects of silibinin on expression of integrin linked kinase, transforming growth factor β1 and a-smooth muscle actin in rat peritoneal mesothelial cells induced by high glucose

作　　者：孙立霞[1] 马健飞[1] 樊怡[1] 杨丽娜[1] 邓文艳[1] 王力宁[1] 

机构地区：[1]中国医科大学附属第一医院肾脏内科,沈阳110001

出　　处：《中华肾脏病杂志》2013年第4期268-272,共5页Chinese Journal of Nephrology

摘　　要：目的研究水飞蓟宾对高糖作用下大鼠腹膜间皮细胞（RPMC）整合素连接激酶（ILK）、转化生长因子β1（TGF—β1）及α平滑肌肌动蛋白（α-SMA）表达的影响。方法胰蛋白酶消化法原代及传代培养RPMC，经鉴定后第2代用于实验。细胞随机分组：（1）正常对照组；（2）高糖组：不同质量浓度的葡萄糖（1．5％、2．5％、4．25％）作用24h；2．5％葡萄糖分别作用RPMC12、24、48、72h；（3）水飞蓟宾组：不同质量浓度的水飞蓟宾（5、10、20mg／L）预孵育2h，加入2．5％葡萄糖再作用24h。实时定量PCR检测ILK及α-SMA mRNA的表达。Western印迹法检测ILK蛋白表达。ELISA法检测细胞培养上清液中TGF—β1的表达。结果与正常对照组相比，高糖可以刺激RPMCILK及α-SMAmRNA的表达增高（均P〈0．05）；高糖诱导RPMCILK及TGF-β1蛋白的表达增高（均P〈0．05）；与高糖组相比，水飞蓟宾能显著降低高糖所致的RPMCILK、TGF-β1及d．SMA的高表达（均P〈0．05）。结论高糖可上调RPMCILK、TGF—β1及α-SMA的表达；水飞蓟宾可以抑制高糖所致的RPMCILK、TGF-β1及α—SMA的高表达。Objective To observe the effect of silibinin on the expression of integrin linked kiuase （ILK）, transforming growth factor β1 （TGF- β1） and α- smooth muscle actin （α- SMA） in rat peritoneal mesothelial cells （RPMCs） induced by high glucose. Methods RPMCs were isolated, cultured and passaged by trypsin, then identified. The second generation of cultured RPMCs were used in the experiment. RPMCs were divided into normal control group, high glucose（1.5%, 2.5%, 4.25%） for 24 hours, high glucose （2.5%） for 12, 24, 48, 72 hours, high glucose （2.5%） for 24 hours after silibinin （5, 10, 20 mg/L） preincubate for 2 hours. ILK and α-SMA mRNA were detected by real-time PCR. ILK protein was detected by Western blotting. TGF-β1 protein in supernatants was detected by ELISA. Results Compared with the control group, the expresssion of ILK, TGF-β1 and α-SAM was significantly increased in groups stimulated by high glucose （all P 〈 0.05）. Silibinin could significantly decrease the expression of ILK, TGF- β1 and α- SMA induced by high glucose （all P 〈 0.05）. Conclusions High glucose can up-regulate the expression of ILK, TGF-β1 and α-SMA. Silibinin can reverse these changes.

关 键 词：细胞转分化 纤维化 腹膜透析 整合素连接激酶 转化生长因子Β1 Α平滑肌肌动蛋白 

分 类 号：R28[医药卫生—中药学]