Objective: Multidrug resistance (MDR) is one main cause of chemotherapy failure. Baicalin is an important active ingredient with anticancer potential in many Chinese herbal medicines. In order to understand the function of baicalin reversing MDR in hepatocellular carcinoma (HCC) and the molecular mechanisms that underlie it, the current study was designed. Methods: Bel-7402 and Bel-7402/5-FU cells were cultured, and MTT assay was applied to detect cell viability and the cross-resistance of Bel-7402/5-FU cells. The pump function, apoptosis, and autophagy were detected by flow cytometry. The related proteins were detected by Western blot assay. The PI3K agonist (740Y-P) was used to verify whether baicalin overcomes the drug resistance of HCC cells by blocking the PI3K/AKT pathway. Results: The findings showed that Bel-7402/5-FU cells were cross-resistant to different chemotherapeutic drugs. Baicalin inhibited cell viability in both Bel-7402/5-FU and Bel-7402 ​cells, and baicalin increased sensitivity of Bel-7402/5-FU cells to 5-FU in time- and dose-dependent manners. Baicalin increased the accumulation of doxorubicin and rhodamine-123 in Bel-7402/5-FU cells and inhibited the protein expression of ABCG2, ABCB1, and ABCC1, associated with pump function. In addition, baicalin induced apoptosis of Bel-7402/5-FU cells via up-regulating Bax expression. Furthermore, baicalin increased autophagy through regulating LC3-Ⅱ, p62, and Beclin-1. Baicalin reversed drug resistance in Bel-7402/5-FU cells by inhibiting the PI3K/AKT pathway, which promoted autophagy and apoptosis to restore chemosensitivity. Conclusion: Baicalin increased accumulation of chemotherapy drugs and induced apoptosis and autophagy in Bel-7402/5-FU cells by inhibiting the PI3K/AKT signaling pathway, that may be the important mechanism by which baicalin reverses the MDR of HCC.

目的 探讨前列宁(QLN)通过调控前列腺MMP-2、TIMP-2降解层粘连蛋白(LN)治疗良性前列腺增生的机制.方法 将50只SPF级6～7周龄雄性SD大鼠采用随机数字表法分为空白组,模型组和低、中、高剂量组,每组 10 只,除空白组外,其余组采用氯胺酮腹腔注射麻醉去势摘除双侧睾丸、1 周后按 5 mg/kg皮下注射丙酸睾酮造模,共28 d,造模期间,空白组、模型组按10 ml/(kg·d)灌胃生理盐水,低、中、高剂量组分别按3 g、6 g、12 g/(kg·d)灌胃QLN,1次/d,连续28 d.末次灌胃后禁食12 h,分离各组大鼠完整的前列腺组织并计算前列腺体积、前列腺湿重及前列腺指数(PI),免疫组化检测各组大鼠前列腺组织LN表达水平;取对数生长期的人前列腺增生细胞-1(BPH-1)分为空白组和低、中、高剂量组,分别按浓度为0、0.25、0.5、1.0 mg/mL QLN干预48 h,倒置显微镜观察各组细胞形态,CCK-8法检测各组细胞活性,Western blot分别检测各组细胞MMP-2、TIMP-2、LN蛋白表达量.结果 与空白组比较,模型组大鼠前列腺体积、前列腺湿重及PI均明显增大(P＜0.01),前列腺组织LN表达水平均明显增强(P＜0.05);与模型组比较,低、中、高剂量组大鼠前列腺体积、前列腺湿重及PI均明显减小(P＜0.05),低、中、高剂量组大鼠前列腺组织LN表达水平均不同程度降低(P＜0.05).与空白组比较,中、高剂量组BPH-1细胞随着浓度的增高,由菱形变成不规则,密度明显减少,生长速度缓慢,24、48 h BPH-1细胞活力均明显下降(P＜0.05),MMP-2、LN蛋白表达量均明显降低,TIMP-2蛋白表达量明显增高(P均＜0.05).结论 QLN能有效减小前列腺体积、前列腺湿重及PI,降低前列腺组织LN表达水平,其机制之一可能是通过调控前列腺组织及BPH-1中MMP-2、TIMP-2降解LN而得以实现.

Background: Our previous studies reported that Qianliening capsule (QC) has a significant therapeutic effect on BPH. Therefore, we investigated the effect QC on apoptosis of human prostatic hyperplasia epithelial-1 cells (BPH-1). Methods: The BPH-1 cells were treated with various concentrations of QC in vitro. Morphology of BPH-1 cell was observed, and the cell viability was determined by the 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay. The levels of Cytochrome C, caspase-9 and caspase-3 were detected using the flow cytometry and colorimetric assay respectively. The Bax mRNA and the miRNA-221, -222, -15a, -16, -181a was determine by Real-time PCR analysis. Results: The apoptosis of BPH-1 cells treated with QC increased than that of untreated cells, as evidenced by loss of plasma membrane asymmetry, the nuclear condensation and fragmentation, collapse of mitochondrial membrane potential in a dose depended manner. The levels of Cytochrome C and caspase-9, caspase-3 in the cells treated with QC increased using the flow cytometry and colorimetric assay respectively. The mRNA and protein expression of Bax and the expression of miRNA-181a in the cells treated with QC increased in a dose dependent manner. Conclusion: QC could induce BPH-1 cells apoptosis by regulating miRNA-181a mediated mitochondrial dependent apoptosis pathway, which may be one of the important mechanisms that QC treated benign pro static hyperplasia. Copyright (C) 2018, Taiwan Society of Geriatric Emergency & Critical Care Medicine. Published by Elsevier Taiwan LLC.

目的:观察前列宁胶囊(QC)对基质金属蛋白酶-2(MMP-2)介导的细胞外基质(ECM)的影响,探讨QC对BPH的治疗机制.方法:SD大鼠随机分为空白组,BPH模型组,QC低、中、高观察组,建立BPH模型,QC干预,ELISA法检测大鼠血清MMP-2、FN、Collagen IV、LN;BPH-1细胞培养,分别加入MMP-2和未加MMP-2刺激因子,QC干预,MTT法观察细胞活性,Q-PCR及Western-Blot分别检测MMP-2、FN、Collagen IV、LN基因及蛋白表达(未加MMP-2),加入MMP-2组检测FN、Collagen IV、LN.结果:QC各剂量组大鼠血清中的FN、LN、Collagen IV含量降低,MMP-2升高(P<0.05 or P<0.01);BPH-1细胞培养QC干预,加入及未加入MMP-2刺激因子细胞活力均有下降,以48 h后最明显;加入MMP-2刺激因子FN、Collagen IV、LN基因及蛋白表达明显低于未加MMP-2刺激因子,差异有统计学意义.结论:QC对BPH具有治疗作用,QC调控MMP-2介导细胞外基质FN、Collagen IV、LN基因和蛋白表达可能是其治疗BPH机制之一.

目的 研究前列宁胶囊治疗良性前列腺增生(BPH)的机制. 方法 用SD大鼠制作BPH模型,光镜观察6组大鼠前列腺组织病理改变,Real-time PCR法检测6组大鼠前列腺组织Fas、Caspase-8、Caspase-3、Bax及bcl-2基因表达,BPH-1细胞培养,前列宁胶囊不同剂量干预,比色法分析Caspase-8、Caspase-3活化,Real-time PCR检测Fas、Bax及bcl-2基因表达,Westen-blot检测Fas、bcl-2、Bax蛋白表达. 结果 病理显示前列腺病理组织明显改善,前列腺组织中Fas、Caspase-8、Caspase-3、Bax基因表达增强,bcl-2表达降低;BPH-1细胞培养,前列宁胶囊干预后,倒置显微镜观察BPH-1细胞胞密度明显减少,细胞变小,细胞凋亡增加,前列宁胶囊不同剂量干预比色法分析Caspase-8、Caspase-3活化增强,Real-time PCR、Westen-blot检测Fas、Bax基因及蛋白表达增强,bcl-2降低. 结论 前列宁胶囊调控Caspase-3通路中的相关因子,促进前列腺组织细胞凋亡,是其治疗BPH的机制.

The present study investigated whether Qianliening capsules (QC) affected the apoptosis of benign prostatic hyperplastia epithelial (BPH‑1) cells by regulating the extracellular matrix (ECM). The levels of fibronectin (FN) and collagen IV were determined in the culture medium of BPH‑1 cells maintained in normal medium and of BPH‑1 cells maintained in an environment rich in FN and collagen IV using an enzyme‑linked immunosorbent assay. Reverse transcription quantitative polymerase chain reaction and western blot analysis were performed to determine the mRNA and protein expression levels of FN, collagen IV, B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and cyclin D1, respectively. The cell morphology and viability were determined using light microscopy and an MTT assay and cell apoptosis was detected by annexin V staining. The results demonstrated that FN and collagen IV affected the apoptotic response of the BPH‑1 cells, QC treatment significantly reduced the levels of FN and collagen IV secreted by the cells into the culture medium (P<0.01), inhibited the mRNA and protein expression levels of FN, collagen IV, Bcl‑2 and cyclin D1 and promoted the mRNA and protein expression of Bax. Therefore, one of the mechanisms underlying the anti‑BPH action of QC involves promoting apoptosis by regulating the expression of the extracellular matrix.

Previous studies by our group showed that Qianliening capsules (QC), a clinically proven effective traditional Chinese formulation that has long been used in the treatment of benign prostatic hyperplasia (BPH), is capable of inhibiting BPH in vivo and in vitro via the promotion of apoptosis, suppression of the EGFR/STAT3 signaling pathway and regulating the expression of sex hormones as well as their receptors. However, the mechanism of its anti-BPH activity has remained to be fully elucidated. The present study aimed to investigate the mechanism underlying the anti-proliferative effect of QC in vivo and in vitro. Castrated male Sprage-Dawley (SD) rats where subcutaneously injected with testosterone propionate and the WPMY-1 cell line was stimulated with basic fibroblast growth factor in order to generate BPH in vivo and in vitro separately, both of which were then subjected to QC treatment. Finasteride was used as a positive control drug for the in vivo study. In the present study, it was found that treatment with QC or finasteride significantly reduced the prostatic index (PI=prostate wet weight/body weight x 100) in a rat model of BPH (P<0.05). In addition, reverse transcription quantitative polymerase chain reaction (RT-PCR) and western blot analyses showed that QC or finasteride treatment significantly inhibited model construction-induced upregulation of expression of proliferating cell nuclear antigen, cyclin D1 and cyclin-dependent kinase 4 in prostatic tissues of rats with BPH (P<0.05). The in vitro study further proved that QC exhibited anti-proliferative properties via G1/S cell cycle arrest in the WPMY-1 cell line, as evidenced by colony formation, flow cytometric cell cycle, immunoblot and RT-PCR analyses. In conclusion, the present study demonstrated that inhibition of cell proliferation via G1/S cell cycle arrest may be one of the underlying mechanisms of the effect of QC on BPH.

Total alkaloids in Rubus aleaefolius Poir (TARAP) is a folk medicinal herb that has been used clinically in China to treat nonalcoholic fatty liver disease (NAFLD) for many years. However, the mechanism of its anti-NAFLD effect is largely unknown. In this study, we developed a NAFLD rat model by supplying a modified high-fat diet (mHFD) ad libitum for 8 weeks and evaluated the therapeutic effect of TARAP in NAFLD rats as well as the underlying molecular mechanism. We found that TARAP could reduce the serum triglycerides (TG), total cholesterol (TC), and low-density lipoprotein (LDL-C) levels and increase the serum high-density lipoprotein (HDL-C) level in NAFLD rats. In addition, TARAP treatment reduced expression of fatty acid synthetase (FAS), and acetyl-CoA carboxylase (ACC) and upregulated the expression of carnitine palmitoyltransferase (CPT). Our results suggest that regulation of lipid metabolism may be a mechanism by which TARAP treats NAFLD.

Angiogenesis plays an important role in the progression and development of benign prostatic hyperplasia (BPH), and has become a promising target for BPH treatment. The hypoxia-inducible factor-1α (HIF-1α) signaling pathway promotes the process of angiogenesis, contributing to the growth and progression of a number of hyperplasia diseases, including BPH. Qianliening capsule (QC) is a traditional Chinese formula that has been used clinically in China to treat BPH for a number of years. Recently, QC was demonstrated to inhibit prostatic cell growth and induce apoptosis in vivo and in vitro via regulating the epidermal growth factor/signal transducer and activator of transcription 3 signaling pathway and mitochondrion-dependent apoptosis pathway. However, the mechanisms underlying the anti-BPH effect remain largely unknown. To further elucidate the mechanism of QC activity in BPH treatment, a rat BPH model established by injecting testosterone following castration was established and the effect of QC on prostatic tissue angiogenesis was evaluated, as well as the underlying molecular mechanisms. QC was shown to reduce the prostatic index in BPH rats, but without affecting the body weight, demonstrating that QC is effective in the treatment of BPH and without apparent toxicity. In addition, QC treatment significantly reduced the intraprostatic microvessel density, indicating antiangiogenesis activity in vivo. In addition, treatment with QC inhibited the expression of HIF-1α in BPH rats, as well as the expression of vascular endothelial growth factor and basic fibroblast growth factor. Therefore, for the first time, the present study hypothesized that QC inhibits angiogenesis in prostatic tissue of BPH rats via the inhibition of the HIF-1α signaling pathway, which may be one of the mechanisms in which QC treats BPH.

The signal transducer and activator of transcription 3 (STAT3) pathway is one of the main growth factor‑mediated signal transduction pathways and is closely associated with the occurrence and development of benign prostatic hyperplasia (BPH). Qianliening capsules (QC) have significant therapeutic effects on BPH; however, the precise mechanism underlying its anti‑BPH activity remains to be elucidated. To further elucidate the molecular mechanism of the therapeutic effect of QC on BPH, the present study used epidermal growth factor (EGF), which has a role in the pathogenesis of BPH, to stimulate the growth of human prostate WPMY‑1 cells and activate the STAT3 pathway in the WPMY‑1 cells. The cell viability was determined using an MTT assay and the cell morphology was observed by phase‑contrast microscopy. Fluorescence activated cell sorting analysis with Annexin‑V/propidium iodide (PI) staining and PI staining were performed to examine cell apoptosis and the cell cycle. The activation of caspase‑9 and ‑3 were evaluated by colorimetric assay. STAT3 phosphorylation and transcriptional activity were detected by western blot analysis and the luciferase gene reporter, respectively. The mRNA and protein expression levels of B‑cell lymhoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax), cyclin D1, cyclin‑dependent kinase 4 (CDK4) and p21 were measured by reverse transcription quantitative polymerase chain reaction and western blot analysis, respectively. In the present study, QC was found to significantly and dose‑dependently inhibit the EGF‑stimulated growth of WPMY‑1 cells, as evidenced by QC‑induced cell -morphological changes and a reduction in cell viability. In addition, QC treatment markedly induced the activation of caspase‑9 and ‑3. QC treatment also inhibited the EGF‑mediated increase of STAT3 phosphorylation levels and transcriptional activity in WPMY‑19 cells, accompanied by downregulation of the expression of Bcl‑2, cyclin D1 and CDK4 and upregulation of the expression of Bax and p21. These results suggested that QC effectively inhibited the proliferation and promoted the apoptosis of human prostate cells via modulation of the STAT3 signaling pathway and its target genes, which is likely to be one of the mechanisms underlying its activity in BPH treatment.

Benign prostatic hyperplasia (BPH) is a pathological overgrowth of the human prostate. It may cause increased resistance to urine flow through the urethra and occasionally kidney damage, bladder stones and urinary tract infections, and therefore affect the quality of life. Qianliening capsule (QC) is a traditional Chinese formula that has been used clinically in China to treat BPH for a number of years. However, the mechanism of its anti-BPH effect remains largely unknown. We evaluated the therapeutic effect of QC in a rat model of BPH, established by the injection of testosterone following castration, and investigated the underlying molecular mechanism of action. We observed that QC treatment significantly and dose-dependently decreased the prostatic volume (PV) and prostatic index (PI; P<0.05 or P<0.01), and ameliorated the histological damage of the prostate tissue in the BPH rats. In addition, treatment with QC inhibited the phosphorylation of signal transducer and activator of transcription 3 (STAT3), as well as the expression of epidermal growth factor (EGF), epidermal growth factor receptor (EGFR), cyclin D1 and Bcl-2. Our results suggest that suppression of the EGF/STAT3 pathway may be one of the mechanisms by which QC treats BPH.

Objective:To observe the clinical effect of Qianlieningn Capsule(QC)on benign prostatic hyperplasia(BPH).Methods:90Cases of BPH were randomly divided into two groups,45cases in the treatment group were given QC and 45cases in the control group were given Finasteride for 12weeks.And the changes of maximum flow rate(MFR),average flow rate(AFR),international prostate symptom scores(IPSS),quality of life index(QOL),the safety or adverse effect in patients of two groups were observed.Results:The changes of MFR,AFR,IPSS and QOL in treatment group had statistics significant difference respectively before and after the treatment.And the safety or adverse effect in treatment group was no significant difference between QC treatment group and control group.Conclusion:QC in the treatment of BPH is safe,effective.

To investigate the molecular mechanisms by which Qianliening Capsule (前列宁胶囊, QC) treats benign prostatic hyperplasia (BPH).

The objective of the study is to evaluate the effect of Qianliening capsule (QLNC) on the expression levels of serum hormones, prostatic estrogen receptor and androgen receptor in benign prostatic hyperplasia (BPH) rats, and investigate the possible molecular mechanisms mediating its anti-BPH activity. Male Sprage-Dawley (SD) rat BPH model was generated. BPH rats were orally treated with different concentrations of QLNC. Blood and the prostatic tissues of animals were obtained. The prostatic weight (PW) and prostatic index (PI) were evaluated; the histopathological changes of prostatic tissue, the levels of serum testosterone and estradiol, the mRNA and protein expression of ER and AR in prostatic tissue were examined by microscopy with hematoxylin and eosin staining HE staining, ELISA, RT-PCR, immunohistochemistry, respectively. Compared to the model group, the PW and PI in all QLNC-treated groups have significantly lower (p<0.05) serum than T/E-2 in QLNC-treated groups which was elevated significantly (p<0.05 or p<0.01). Pathomorphism of prostatic tissue in QLNC-treated groups improved. The mRNA and protein expression of ER and AR in QLNC-treated groups decreased significantly. QLNC has significant therapeutic effect on BPH rats. Improvement of sex hormones disorder and regulation of ER and AR are one of the mechanisms by which QLNC treats BPH.

OBJECTIVE:To investigate the molecular mechanisms by which Qianliening Capsule (, QC) treats benign prostatic hyperplasia (BPH). METHODS:Human prostate stromal cell line WPMY-1 was treated with 0, 1, 3 and 5 mg/mL of QC for 24, 48 and 72 h, respectively, in the presence of 10 ng/mL basic fibroblast growth factor (bFGF). The viability of WPMY-1 cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell morphology was observed by phase-contrast microscopy. 4',6-diamidino-2-phenylindole (DAPI) staining and fluorescence activated cell sorting (FACS) analysis with Annexin-V/propidium iodide (PI) staining were performed to determine cell apoptosis. The loss of mitochondrial membrane potential was examined by FACS analysis with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyarine iodide (JC-1) staining. Activation of caspase-3 and -9 was evaluated by colorimetric assay. The mRNA and protein expression levels of Bcl-2 and Bax were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. RESULTS:Upon bFGF stimulation, the viability of WPMY-1 cells was increased to 122%-118% compared with the control cells (P <0.05). However, treatment with 1-5 mg/mL of QC for 24, 48 and 72 h decreased the viability of bFGF-stimulated cells to 80%-92%, 59%-82%, 36%-62% compared with the untreated cells (P <0.05). In addition, QC treatment reduced WPMY-1 cell density in a dose-dependent manner. Moreover, QC treatment dose-dependently induced the loss of plasma membrane asymmetry, the nuclear condensation and fragmentation, collapse of mitochondrial membrane potential, activation of caspase-9 and caspase-3, and increase of pro-apoptotic Bax/Bcl-2 ratio. CONCLUSION:Promoting mitochondrion-dependent apoptosis of prostate stromal cells might be one of the mechanisms by which QC treats BPH.

Objective: To reseach the preparation process and its quality standard of Qianliening Enteric-coated Capsules,meanwhile,inspect the release of emodin in vitro.Methods: HPLC was used for determining the content of emodin in compound,and further investigation of the in vitro release.Results: The drug release of the artificial gastric juice of Qianliening Enteric-coated Capsules would be lower than 10%,but in simulated human gastroenteric environment can rapidly release,accumulate more than 90% after 50min later.Conclusion: The parameters of technology and formulation are reasonable and stable.The quality standard is reliable.So it may be a good reference for the other enteric-coated capsules.

We aimed to examine the effect of an alkaloid extract of the roots of Rubus alceifolius Poir on liver damage and cytochrome enzymes, and underlying mechanism. Hepatotoxicity was induced in rats by treatment with carbon tetrachloride (CCl(4)). Rats were then treated with the hepatoprotective drug bifendate, or with low, medium, and high doses of an alkaloid extract from the roots of R alceifolius Poir. Both bifendate and alkaloid treatment decreased the increase in liver enzymes and cell damage caused by CCl(4). Carbon tetrachloride treatment alone caused a decrease in total cytochrome P450 content, an increase in CYP2E1 and CYP3A1 messenger RNA (mRNA) levels, and an increase in CYP2E1 and a decrease in CYP3A1 enzymatic activity. Alkaloid treatment brought these concentrations and activities back toward normal. In summary, these results suggest that alkaloids from R alceifolius Poir may act to protect the liver through decreasing CYP2E1 enzymatic activity through decreasing its mRNA.

Objective:To preliminary research extract preparation and the quality control of Qianliening enteric-coated capsule.Method:Precipitation was used to the preparation and the mass identification was by TLC method.Results:The preparation was stabile and the TLC spots developed were fairly clear.Conclusion:The method is simple,reliable,and can be used to control the quality of Qianliening enteric-coated capsule effectively.

目的 观察前列宁胶囊对良性前列腺增生模型大鼠前列腺组织中EGF、EGFR表达的影响.方法 将雄性SD大鼠60只,随机分成空白组、模型组、保列治组、前列宁胶囊(低、中、高剂量)组;去势后皮下注射丙酸睾酮复制大鼠前列腺增生模型,连续给药28 d,观察各组前列腺涅重及指数,RT-PCR检测大鼠前列腺组织中EGF、EGFR的mRNA表达,免疫组化法观察前列腺组织EGF及EGFR的蛋白表达情况.结果 模型组前列腺湿重、指数明显增加,和正常组相比有显著差异性(P<0.01),前列腺组织EGF及EGFR的mRNA和蛋白表达较正常组均显著升高(P<0.01);和模型组相比,前列宁胶囊各剂量组和保列治组的前列腺湿重与指数明显降低(P<0.05),前列腺组织EGF及EGFR的mRNA和蛋白表达与模型组相比均显著降低(P<0.05).结论 前列宁胶囊对BPH大鼠有明显的治疗作用,能显著地抑制BPH大鼠EGF及EGFR的表达,提示这可能是前列宁胶囊治疗BPH的重要机制之一.

目的 观察前列宁胶囊对实验性良性前列腺增生(BPH)模型大鼠的血清酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性的影响.方法 将雄性SD大鼠60只,随机分成空白组、模型组、保列治组及前列宁胶囊低、中、高剂量组.采用丙酸睾酮皮下注射复制大鼠BPH模型,连续给药28 d后,观察6组前列腺指数、ACP及AKP的变化.结果 模型组大鼠的前列腺指数和血清ACP、AKP水平显著升高,与空白对照组比较有显著差异(P<0.01);前列宁胶囊各剂量组的前列腺指数和ACP、AKP活性均有降低,其中各药物治疗组的前列腺指数和AKP活性与模型组比较具有显著性差异(P<0.05),但前列宁胶囊低荆量组引起的ACP变化没有显著性差异(与模型组比较,P>0.05),而中剂量组和高剂量组对血清ACP水平的抑制作用具有非常显著性差异(P<0.01).结论 前列宁胶囊具有抑制BPH大鼠的前列腺指数及降低血清ACP和AKP活性的作用.