OBJECTIVEThe prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.

METHODSPrimary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.

RESULTSThe Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).

CONCLUSIONSiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.

Apoptosis ; genetics ; Apoptosis Regulatory Proteins ; genetics ; Bone Marrow Cells ; cytology ; metabolism ; Caspase 3 ; metabolism ; Cells, Cultured ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Small Interfering

Objective: To investigate the effects of transrectal ultrasound conductance (TRUSC)-guided administration of traditional Chinese medicine on histological prostatitis in men with small-size BPH and low urinary tract symptoms (LUTS) after transurethral resection of the prostate (TURP). METHODS: This study included 167 BPH patients without surgical contraindications. We randomized the patients into an experimental group (n = 84) and a control group (n = 83), with no statistically significant differences between the two groups in age, prostate volume, International Prostate Symptom Score (IPSS), and quality of life (QoL) (P >0.05). The patients of the experimental group received TRUSC-guided administration of traditional Chinese medicine, qd, for 7 days before TURP, while those of the control group underwent TURP only. After treatment, we compared the results of postoperative pathological examination of the prostate tissue, the histological grade of inflammation, IPSS, and QoL scores between the two groups of patients. RESULTS: In the experimental group, there were 12 cases of non-inflammation (14.3%), 43 cases of mild inflammation (51.2%), 28 cases of moderate inflammation (33.3%), and 1 case of severe inflammation (1.2%), as compared with 8 cases of non-inflammation (9.6%), 28 cases of mild inflammation (33.7%), 45 cases of moderate inflammation (51.8%), and 2 cases of severe inflammation (2.4%) in the control group (P <0.05). Compared with the baseline, both the experimental and control groups showed significant improvement at 4 weeks after surgery in IPSS (22.20±4.14 vs 4.26±2.64 and 23.05±4.11 vs 7.02±4.15, P <0.05) and QoL scores (4.33±0.83 vs 1.25±1.64 and 4.25±0.91 vs 2.05±1.95, P <0.05). CONCLUSIONS TRUSC-guided administration of traditional Chinese medicine can significantly alleviate histological inflammation and improve QoL in men with small-size BPH and LUTS after TURP.
Drugs, Chinese Herbal ; administration & dosage ; Humans ; Lower Urinary Tract Symptoms ; drug therapy ; Male ; Medicine, Chinese Traditional ; methods ; Prostatic Hyperplasia ; drug therapy ; pathology ; Prostatitis ; drug therapy ; pathology ; Quality of Life ; Transurethral Resection of Prostate ; Treatment Outcome ; Ultrasonography, Interventional ; methods

Objective::To study the chemical constituents from n-butanol extract of Akebia trifoliata caulis. Method::The 100 kg caulis of A. trifoliata was extracted with 75% ethanol (EtOH) for three times by heating reflux. These 3 extracts were decompressed and concentrated, and then dissolved in water. The solvent was successively extracted with dichloromethane, ethyl acetate and n-butanol. The chemical constituents from the n-butanol fraction were isolated by macroporous, silica gel, sephadex LH-20 and ODS columns, and semi-preparative high performance liquid chromatography, and their chemical structures were determined through MS, NMR analysis (¹H and ¹³C-NMR) and spectroscopic data from literatures. Result::Totally 14 compounds were isolated and identified as mutongsaponin B(1), mutongsaponin C(2), saponin PH(3), begoniifolide A(4), 2α, 3β, 23-trihydroxy-30-noroleana-12, 19-dien-28-oicacid-O-β-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester(5), akemisaponins D(6), akemisaponins E(7), asiaticoside(8), saponin PJ1(9), scheffoleoside A(10), symplocosneolignan A(11), kalopanax-saponins D(12), leonticin E(13), ciwujianoside A₁(14). Conclusion::Compounds 1-4, 11, 13, 14 were isolated from this plant for the first time. The discovery of these compounds further enriched the chemical constituents of A. trifoliata, and provided experimental and scientific basis for the comprehensive development and utilization of A. trifoliata.