抗纤灵对5/6肾切除大鼠骨髓来源的成纤维细胞表型转化的影响

Effect of Kangxianling on Phaenotype Transformation of Bone Marrow-derived Fibroblast in 5/6 Nephrectomy Rats

目的观察抗纤灵对5/6肾切除大鼠骨髓来源的成纤维细胞表型转化的影响,探讨其抗肾纤维化的作用机制。方法将SD大鼠随机分为假手术组和造模组,造模组以5/6肾切除方法建立慢性肾衰竭动物模型,1周后按血肌酐水平将造模大鼠分为模型组、福辛普利组和抗纤灵组。各给药组给予相应药物灌胃,模型组和假手术组给予等量生理盐水灌胃,8周后采用实时荧光定量PCR和蛋白免疫印迹法测定各组大鼠肾组织α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原和CD45基因及蛋白的表达。结果模型组大鼠表达α-SMA mRNA、Ⅰ型胶原mRNA和CD45mRNA水平较假手术组明显升高(P<0.001或P<0.05),抗纤灵组与模型组比较均明显下降(P<0.01或P<0.05);模型组大鼠表达α-SMA、Ⅰ型胶原和CD45蛋白水平较假手术组明显升高(P<0.001),抗纤灵组与模型组比较均显著下降(P<0.001)。结论抗纤灵可降低Ⅰ型胶原和CD45的表达,抑制骨髓来源的成纤维细胞表型转化,从而延缓肾纤维化进程,保护残肾功能。

Objective To observe the effect of Kangxianling on phaenotype transformation of bone marrow-derived fibroblast in 5/6 nephrectomy rats, and to explore its mechanism of anti-renal fibrosis. Methods SD rats were randomly divided into sham operation group and model group. The model group was made rat model of chronic renal failure with 5/6 nephrectomy. One week later, the model group rats were randomly divided into untreated group, fosinopril group and Kangxianling group by the serum creatinine level. The fosinopril group was given 3.3 mg/（kg ·d） fosinopril, Kangxianling group was given 23 g/（kg·d） Kangxianling untreated group and sham operation group were given normal saline by gavage for 8 weeks. The expression of α-SMA, collagen Ⅰ and CD45 in the renal tissue were determined by real-time quantitative PCR （RT-PCR） and protein immunoblot method （Western blot）. Results Compared with the sham operation group, the expression of α-SMA mRNA, collagen Ⅰ mRNA and CD45 mRNA levels in the model group were significantly higher （P〈0.001 or P〈0.05）, and those in Kangxianling group were significantly decreased （P〈0.01 or P〈0.05）. The expression of α-SMA, collagen Ⅰ and CD45 levels in the model group were significantly higher than that in the sham operation group, and those in Kangxianling group were significantly decreased （P〈0.001）. Conclusion Kangxianling could depress the expression of collagen Ⅰ and CD45, suppress phaenotype transformation of bone marrow-derived fibroblast, thereby delay the process of renal fibrosis and protect residual renal function.