前癃通胶囊含药血浆对前列腺间质细胞Smad4基因表达的影响

目的:探讨中药前癃通胶囊含药血浆对体外培养前列腺间质细胞Smad4基因表达的影响,为中药复方治疗良性前列腺增生(BPH)提供实验依据。方法:收集2012年8~10月门诊BPH患者15例,按照随机数字表对入选患者进行实验分组,分为前癃通高、中、低剂量组,制备前癃通含药血浆。同时,体外培养前列腺间质细胞,当细胞接近长成单层后进行免疫荧光检测,进行间质细胞鉴定;加含药血浆后培养24 h,采用实时荧光定量PCR(q-PCR)技术检测间质细胞Smad4基因表达;Western印迹检测Smad4蛋白在间质细胞的表达。结果:经含药血浆处理后间质细胞Smad4基因mRNA的表达随药物剂量增加依次增强,与空白组、无血浆组比较,各组差异均有统计学意义(P0.01)。经含药血浆处理后间质细胞Smad4蛋白的表达随药物剂量增加依次增强,与空白组比较,低剂量组差异无统计学意义(P0.05),中剂量组差异有统计学意义(P0.01);与低剂量组比较,中剂量组差异有统计学意义(P0.05);与高剂量组比较,各组之间差异均有统计学意义(P均0.01)。结论:中药前癃通胶囊含药血浆体外培养前列腺间质细胞,可抑制其增殖,其机制与促进Smad4基因及蛋白表达有关。

Qianlongtong Capsule elevates the Smad4 gene expression in prostate stromal cells

Objective： To investigate the effects of the plasma containing Qianlongtong Capsule （QLT）-containing plasma on the expression of the Smad4 gene in prostate stromal cells in vitro and provide some experimental evidence for the treatment of benign prostatic hyperplasia （BPH） with Chinese medicinal compound. Methods： Fifteen cases of BPH were equally randomized to three groups to be treated with QLT at a high dose （6 capsules once）, a medium dose （3 capsules once）, and a low dose （1.5 capsules once）, tid, for 7 days consecutively. QLT-containing plasma was collected from the patients. Prostate stromal cells were identified by immunofluorescence when they became monolayered and cultured in the QLT-containing plasma for 24 hours, followed by detection of the expression of the Smad4 gene by real-time quantitative PCR and that of the Smad4 protein by Western blot. Results： After treat- ment with the QLT-containing plasma, the expression of the Smad4 gene in the stromal ceils was significantly increased in a dose-de- pendent manner as compared with the blank control and no-QLT groups （ P 〈 0.01 ）. The expression of the Smad4 protein was also markedly elevated after treatment. The differences were statistically significant between the blank control and medium-dose groups （P 〈 0.01 ）, low-dose and medium-dose groups （P 〈 0.05 ）, and high-dose and the other groups （P 〈 0.01 ）, but not between the blankcontrol and low-dose groups （P 〉 O. 05 ）. Conclusion ： QLT-containing plasma could inhibit the proliferation and improve the apop- totic index of prostate stromal cells in vitro, which was related to the elevation of the mRNA and protein expressions of Smad4.