“慢性肾衰基本方”含药血清对人肾小管上皮细胞转分化的影响  

The Effect of “Chronic Kidney Failure Basic Prescription” Containing Sera on Human Renal Tubular Epithelial Cells Transdifferentiation

作　　者：刘华锋[1] 洪涛[1] 吴育思[2] 姚翠微[1] 梁东[1] 唐德燊[1] 陈孝文[1] 

机构地区：[1]广东医学院附属医院肾病研究所,湛江524001 [2]广东医学院附属医院药剂科,湛江524001

出　　处：《中国中西医结合肾病杂志》2006年第6期320-323,373,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：广东省中医药管理局基金资助项目(No.103073)

摘　　要：目的:观察经验方“慢性肾衰基本方”对转化生长因子β1(TGF-β1)所诱导的肾小管上皮细胞(TECs)转分化及细胞外基质分泌的影响,以进一步证实并初步探讨其延缓慢性肾衰竭肾小管-间质纤维化的作用机制。方法:给予家兔中药灌胃3d后制备“慢性肾衰基本方”含药兔血清,应用不同浓度含药血清处理经TGF-β1诱导活化的HK-2细胞,然后应用免疫细胞化学方法、流式细胞技术测定HK-2细胞α-平滑肌肌动蛋白(α-SMA)的表达率;应用酶联免疫吸附检测法(ELISA)测定培养上清纤维连结蛋白(FN)及Ⅳ型胶原(Col-Ⅳ)水平,并进而计算单个细胞FN及Col-Ⅳ分泌水平。结果:含药兔血清呈剂量依赖方式逆转TGF-β1诱导的HK-2细胞α-SMA的表达,并呈剂量依赖方式抑制单个细胞FN及Col-Ⅳ的分泌。结论:“慢性肾衰基本方”至少部分是通过抑制肾小管上皮细胞转分化及细胞外基质的过度分泌而对防治肾小管-间质纤维化发挥作用。Objective： To observe the effects of ＂chronic kidney failure basic prescription＂ on the transdifferentiation and extracellular matrix（ECM）secretion in tubular epithelial cells induced by transforming growth factor beta （TGF- β1 ） ,so as to further prove the effects of our empirical prescription and to explore its effective mechanism of anti renal tubulointerstitial fibrosis during chronic kidney failure. Methods： Rabbit sera containing herbs were collected after rabbits were intragastriely administered with the herbs. HK - 2 cells activated by TGF - β1 were incubated with heat - inactivated sera containing herbs in different concentration. The expression of α - SMA in HK - 2 cells was detected by immunocytochemistry and its expression rate was measured by flow cytometry. The fibronectin （FN） and collagen IV （Col- Ⅳ ） secreted in culture supernatant were also measured by enzyme linked immunoassay（ELISA） and then the secretion amount of these two ECM in single cell level was calculated. Results： Rabbit sera containing herbs dosage - dependently reversed the expression of α-SMA induced by TGF-β1 in HK-2 cells, as well as dosage- dependently reduced the secretion of FN and Col- Ⅳ in single cell level. Conclusion： ＂Chronic kidney failure basic prescription＂exerts its protecting effects on renal tubulointerstitial fibrosis partly through inhibiting cells transdifferentiation and further reducing over production of ECM in renal TECs.

关 键 词：慢性肾衰基本方 肾小管-间质纤维化 肾小管上皮细胞转分化 细胞外基质 

分 类 号：R285.5[医药卫生—中药学]