造血细胞生长抑制因子抑制大鼠肾间质成纤维细胞的生长

Antiproliferative Effects of the Hemoregulatory Inhibitor N-acetyl-seryl-aspartyl-lysyl-proline on Rat Renal Interstitial Fibroblasts

摘要:

目的：观察一种新发现的生理性造血细胞生长抑制因子N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸N-acetyl-scryl-aspartyl-lysyl-proline-(AcSDKP)是否对大鼠肾间质成纤维细胞也具有一定程度的生长抑制作用。方法：应用成纤维细胞培养技术，将大鼠肾间质成纤维细胞在不同浓度的AcSDKP中共同孵育24，48 h后，测定其alamar blue的OD吸收值，观察不同浓度的AcSDKP对大鼠肾间质成纤维细胞的生长抑制作用。 结果：大鼠肾间质成纤维细胞在AcSDKP中培养48 h后，测定其alamar blue的OD吸收值，发现其吸收率与对照组相比差异有显著性意义(P<0.01)。说明AcSDKP确实对大鼠肾间质成纤维细胞具有一定程度的生长抑制作用，并且其抑制作用在一定浓度范围内呈现剂量依赖性，当AcSDKP的浓度高于10-5 mol/L或低于10-8 mol/L时，其抑制作用消失，最大抑制浓度为10-6 mol/L，最大抑制率约33%(10-6 mol/L)。结论：生理性造血细胞生长抑制因子AcSDKP的抑制生长作用并不仅仅局限于造血细胞，它对大鼠肾间质成纤维细胞也具有生长抑制作用。

Abstract:

OBJECTIVE: To determine whether the tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP), a physiological hemorcgulatory inhibitor, has a biological function in nonhematopoietic cells (rat renal interstitial fibroblasts). METHODS: Cell proliferation was measured using standard cell culture techniques and the alama blue colorimctric assay. RESULTS: OD measurement of alamar blue absorption at 48 h was significantly inhibited in fibroblasts exposed to 1 μmol AcSDKP (P<0.01). During the 48hour alamar blue absorption, only AcSDKP in concentrations of 10-7 mol/L, 10-6 mol/L, and 10-5 mol/L had significant inhibiting effects compared to cells grown in the 5% FBS RPMI 1640 media alone. The inhibition was dose dependent, and disappeared at AcSDKP concentrations higher than 10-5 mol/L or lower than 10-8 mol/L. Maximum inhibition occurred at an AcSDKP concentration of 10-6 mol/L (33% suppression). CONCLUSIONS: The inhibiting effect of AcSDKP is not limited to hematoposis cells. AcSDKP is specifically cleaved to an inactive form by the N terminal active site of angiotensin converting enzyme (ACE) and ACE inhibitors (ACEI) can increase the concentration of AcSDKP. It suggests that AcSDKP may be an endogenous modulator of renal cell proliferation and the antiproliferative action of ACEI in renal diseases may be partly mediated by AcSDKP.