丹参干预成纤维细胞增生及其机制

Preventive effect and mechanism of Salviae miltiorrhizae on fibroblast proliferation

目的观察丹参对结膜成纤维细胞增生的抑制作用并探讨其作用机制,为临床防治PVR提供新的治疗方法。方法以体外培养的兔结膜成纤维细胞为研究对象,以地塞米松、5-氟尿嘧啶、氢化可的松作为处理对照组,用MTT法测定不同浓度丹参作用48h后成纤维细胞增生情况。AnnexinV-FITC和碘化丙啶双染法,在流式细胞仪下测定不同浓度丹参作用不同时间后成纤维细胞的早期凋亡率。用末端脱氧核糖核酸酶介导的dUTP末端标记法(TUNEL法)观察丹参作用48h后的细胞凋亡情况。结果丹参在一定浓度范围内(1.74～111.00g·L-1)可抑制成纤维细胞增生,且呈剂量依赖关系(0.91%～79.28%)。3.47g·L-1、27.75g·L-1及111.00g·L-1丹参作用6h时细胞凋亡率分别为(8.67±0.97)%、(11.04±1·04)%、(20.83±1.74)%,作用24h时分别为(13.06±1.41)%、(25.68±1.21)%、(39.65±0.23)%,细胞早期凋亡率与正常对照组(4.40±0·08)%相比差异均有统计学意义(均为P<0.01)。丹参作用48h后,成纤维细胞核出现阳性着色。结论丹参可以通过诱导成纤维细胞凋亡的方式来抑制成纤维细胞的增生,有望成为新的防治PVR药物。

Objective To investigate the inhibitive effect and mechanism of Salviae miltiorrhizae on fibroblast proliferation,and provide new medicine for the prevention of proliferative vitreoretinopathy.Methods Using 5-Fu,dexamethasone and hydrocortisone as contrast,the rabbit conjunctival fibroblast cultured in vitro was treated with Salviae miltiorrhizae.After 48 hours of incubation with medicine above,the proliferative alteration of fibroblast was detected by MTT.After different time points of incubation with Salviae miltiorrhizae with different concentrations,the apoptotic rate of fibroblast was detected by flow cytometry.After 48 hours of incubation with Salviae miltiorrhizae,the apoptosis of cultured conjunctival fibroblast was determined by TUNEL method.Results Salvia miltiorrhizae with certain concentrations（from 1.74 g·L-1 to 111.00 g·L-1） could inhibit the proliferation of in a dose-dependent manner.The apoptotic rate of Salviae miltiorrhizae with 3.47 g·L-1,27.75 g·L-1 and 111.00 g·L-1 on fibroblast at 6 hours were（8.67±0.97）%,（11.04±1.04）% and（20.83±1.74）%,which at 24 hours were（13.06±1.41）%,（25.68±1.21）% and（39.65±0.23）%,respectively;Compared with the contrast group（4.40±0.08）%,there were significant differences（all P0.01）.After 48 hours of incubation with different concentrations of Salviae miltiorrhizae,the nucleus of fibroblast showed positive staining.Conclusion Salviae Miltiorrhizae inhibit the proliferation of fibroblast by inducing their apoptosis,and has the potential role in treating proliferative vitreoretinopathy.