抗纤灵方药物血清对骨髓来源成纤维细胞表型转化的影响

Effect of Kangxianling on Phenotypic Transformation of Bone Marrow-Derived Fibroblast

目的探讨抗纤灵方对肾间质纤维化(RIF)的作用机制。方法将抗纤灵方煎至含生药3.2g/ml、福辛普利0.33mg/ml给大鼠灌胃,正常组给予等量蒸馏水灌胃,分别制备抗纤灵方血清、福辛普利血清和正常血清。用DMEM培养基稀释血清,分为正常血清组、福辛普利组、抗纤灵组、转化生长因子-β1(TGF-β1)组、TGF-β1+福辛普利组和TGF-β1+抗纤灵组。体外培养大鼠肾组织骨髓来源的成纤维细胞(FBS),采用酶联免疫吸附法和双重荧光定量法检测骨髓来源的FBS表型转化标记物α-平滑肌肌动蛋白(α-SMA)及Ⅰ型胶原(CollagenⅠ)及其mRNA表达。结果 TGF-β1组α-SMA、CollagenⅠ及α-SMAmRNA、CollagenⅠmRNA表达水平较正常血清组明显增强(P<0.05或P<0.01),TGF-β1+福辛普利组和TGF-β1+抗纤灵组α-SMA、CollagenⅠ及α-SMAmRNA、CollagenⅠmRNA表达水平均低于TGF-β1组(P<0.05或P<0.01)。结论抗纤灵方药物血清可以通过抑制骨髓来源的FBS的表型转化拮抗RIF。

Objective To study the effect and mechanism of Kangxianling on renal interstitial fibrosis （RIF）. Methods Kangxianling serum, fosinopril serum and normal serum was prepared respectively with intragastric administration of 3.2 g/ml Kangxianling, 3.3 mg/mI fosinopril and distilled water to rats. The serum was diluted with a DMEM medium and divided into the normal group, fosinopril group, Kangxianling group, transforming growth factor-β1 （TGF-β1） group, TGF-β1 ＋ fosinopril group and TGF-β1 ＋Kangxianling group. The bone marrow-derived fetal bovine serum （FBS） was studied. The expressions of a-smooth muscle actin （a-SMA） and Collagen I were examined by enzyme-linked immunosorbent assay （ELISA） and real time polymerase chain reaction （RT-PCR）. Results Comparing with the normal group, the expressions of a-SMA, Collagen I, a- SMA mRNA and Collagen I mRNA were significantly improved in the TGF-β1 group （P〉0.05 or P〈0.01）. Comparing with the TGF-β1 group, the expressions of a-SMA, Collagen I, a-SMA mRNA and Collagen I mRNA were significantly decreased in the TGF-β1t＋fosinopril group and TGF-β1＋Kangxianling group （P〈0.05 or P〈0. 01）. Conclusion fKangxianling can antagonize RIF by inhibiting phenotypic transformation of bone marrow-derived FBS.