麝香保心丸防治阿霉素心脏毒性的临床研究

目的:观察麝香保心丸防治阿霉素心脏毒性的临床疗效.方法:选取2014年5月至2016年4月在我科行化疗的恶性肿瘤患者共50例,随机分为观察组和对照组各25例,两组均使用含阿霉素的方案进行化疗,观察组在上述基础上加用麝香保心丸(2粒/次,每天3次,温水吞服,化疗期间及化疗间歇期间均坚持服用),于化疗第3个周期结束后及第6个周期结束后分别接受左室射血分数(LVEF)、短轴缩短率(FS)、舒张早期与晚期充盈速度比值(A/E)、肌钙蛋白(cTnⅠ)、肌酸激酶同工酶(CK-MB)、B型尿钠肽(B-BNP)检测.结果:第6个周期化疗结束后,观察组的LVEF、FS、A/E的降低幅度均较对照组低,与对照组比较差异均有统计学意义(P＜0.05);观察组的cTnⅠ、CK-MB、B-BNP值升高幅度较对照组低,与对照组比较差异均有统计学意义(P＜0.05).结论:麝香保心丸可以明显减轻阿霉素的心脏毒性,值得在临床中进一步研究和运用.     

Anti-inflammatory agents and monoHER protect against DOX-induced cardiotoxicity and accumulation of CML in mice

Cardiac damage is the major limiting factor for the clinical use of doxorubicin (DOX). Preclinical studies indicate that inflammatory effects may be involved in DOX-induced cardiotoxicity. Nepsilon-(carboxymethyl) lysine (CML) is suggested to be generated subsequent to oxidative stress, including inflammation. Therefore, the aim of this study was to investigate whether CML increased in the heart after DOX and whether anti-inflammatory agents reduced this effect in addition to their possible protection on DOX-induced cardiotoxicity. These effects were compared with those of the potential cardioprotector 7-monohydroxyethylrutoside (monoHER).BALB/c mice were treated with saline, DOX alone or DOX preceded by ketoprofen (KP), dexamethasone (DEX) or monoHER. Cardiac damage was evaluated according to Billingham. Nepsilon-(carboxymethyl) lysine was quantified immunohistochemically.Compared to saline, a 21.6-fold increase of damaged cardiomyocytes was observed in mice treated with DOX (P<0.001). Addition of KP, DEX or monoHER before DOX significantly reduced the mean ratio of abnormal cardiomyocytes in comparison to mice treated with DOX alone (P相似文献   

A comparative study between catalase gene therapy and the cardioprotector monohydroxyethylrutoside (MonoHER) in protecting against doxorubicin-induced cardiotoxicity in vitro

Cardiotoxicity is the main dose-limiting side effect of doxorubicin in the clinic. Being a free radical producer, doxorubicin affects the heart specifically because of its low antioxidant capacity. Among those antioxidants, catalase is present in very low levels in the heart compared to other organs. Since catalase is an essential enzyme in detoxifying hydrogen peroxide, the aim of the present study was to investigate the protective effect of catalase as delivered by an adenovirus vector against doxorubicin-induced cardiotoxicity in cultured neonatal rat cardiac myocytes (NeRCaMs). 7-Monohydroxyethylrutoside (MonoHER), a potent cardioprotector currently under clinical investigations, was included in the study as a reference. Neonatal rat cardiac myocytes were infected with different multiplicity of infections (MOIs) of adenovirus encoding catalase (AdCat). A control infection with an adenovirus vector encoding a nonrelated protein was included. The activity and content of catalase in infected cells were determined during 3 days postinfection. One group of NeRCaMs was infected with AdCat before treatment with doxorubicin (0-50 microM). The second and third group were treated with doxorubicin (0-50 microM) with and without 1 mM monohydroxyethylrutoside (monoHER), respectively. The LDH release and viability of treated cells were measured 24 and 48 h after doxorubicin treatment. The beating rate was followed in three other groups of cells receiving the same treatments within 3 days after doxorubicin (0-100 microM) treatment. Catalase activity increased in AdCat-infected cells, with different MOIs, starting from the second day after infection as compared to the mock-infected cells (P<0.03). At the third day of infection, an MOI of more than 50 caused cytopathic effects, which hampered the use of higher viral titres. With an MOI of 50, catalase activity increased 3.5-fold in AdCat-infected cells 3 days postinfection (P=0.021) compared to mock-infected cells. The beating rate and survival of NeRCaMs decreased in a concentration and time-dependent manner after doxorubicin treatment (P<0.0005). This cytotoxicity was associated with an increase in the LDH release from the treated cells (P<0.0005). The cells stopped beating 24 h after treatment with >50 microM doxorubicin. A 3.5-fold increase in the activity of catalase did not protect NeRCaMs against any of the cytotoxic effects of doxorubicin on NeRCaMs. In contrast, monoHER (1 mM) significantly protected NeRCaMs against the lethal effects of doxorubicin on the survival, LDH release and the beating rate of NeRCaMs (P<0.004) during 48 h after doxorubicin treatment. This protection resulted in a prolongation of the beating of doxorubicin-treated cells after the end of the experiment (i.e. >72 h). The present study (1) illustrates that the cytotoxicity of high MOI of AdCat (>50) limited the possibility to increase catalase activity more than 3.5-fold, which was not enough to protect infected NeRCaMs against doxorubicin-induced cardiotoxicity and (2) confirms the efficacy of monoHER as a cardioprotector. Thus, the use of monoHER proves more suitable for the prevention of doxorubicin-induced cardiotoxicity than catalase gene transfer employing adenovirus vectors.     

The effect of monohydroxyethylrutoside on doxorubicin-induced cardiotoxicity in patients treated for metastatic cancer in a phase II study

The purpose of this study was to investigate the cardioprotective effect of the semisynthetic flavonoid 7-monohydroxyethylrutoside (monoHER) on doxorubicin (DOX)-induced cardiotoxicity in a phase II study in patients with metastatic cancer. Eight patients with metastatic cancer were treated with DOX preceded by a 10 min i.v. infusion of 1500 mg m(-2) monoHER. Five patients were examined by endomyocardial biopsy after reaching a cumulative dose of 300 mg m(-2). Histopathological changes in the cardiomyocytes (Billingham score) were compared with those described in literature for patients treated with DOX only. The mean biopsy score of the patients was higher (2.7) than the mean score (1.4) of historical data of patients who received similar cumulative doses of DOX. Although there is a considerable variability in few investigated patients, it was indicative that monoHER enhanced DOX-induced cardiotoxicity. However, the antitumour activity of DOX seemed better than expected: three of the four patients with metastatic soft-tissue sarcoma had a partial remission and the fourth patient stable disease. It is likely that the relatively high dose of monoHER is responsible for the lack of cardioprotection and for the high response rate in patients with soft-tissue sarcoma possibly by depleting the glutathione defense system in both heart and tumour.     

The anticancer prodrugs of butyric acid AN-7 and AN-9, possess antiangiogenic properties

The antiangiogenic and antineoplastic activities of the butyric acid prodrugs AN-7 and AN-9 were demonstrated in vitro with HUVEC by inhibition of proliferation and vascular tubes formation, enhanced apoptosis, and inhibition of 22Rv-1 cells migration. In the sc implanted human prostate tumors (22Rv-1) in nude mice, AN-7 significantly inhibited Ki-67, HIF-1alpha, HER-2/neu, bFGF and increased PTEN level. AN-7 and AN-9 reduced hemoglobin accumulation in matrigel plugs implanted sc in Balb-c mice. Herein, we show that the anticancer activity of AN-7 and AN-9 can be attributed in part to their antiangiogenic activities suggesting potential therapeutic benefits for prostate cancer patients.     

Ursodeoxycholic acid modulates histone acetylation and induces differentiation and senescence

Agents that can modulate colonic environment and control dysregulated signaling are being evaluated for their chemopreventive potential in colon cancer. Ursodeoxycholate (UDCA) has shown chemopreventive potential in preclinical and animal models of colon cancer, but the mechanism behind it remains unknown. Here biological effects of UDCA were examined to understand mechanism behind its chemoprevention in colon cancer. Our data suggests that UDCA can suppress growth in a wide variety of cancer cell lines and can induce low level of apoptosis in colon cancer cells. We also found that UDCA treatment induces alteration in morphology, increased cell size, upregulation of cytokeratin 8, 18 and 19 and E-cadherin, cytokeratin remodeling and accumulation of lipid droplets, suggesting that UDCA induces differentiation in colon carcinoma cells. Our results also suggest significant differences in UDCA and sodium butyrate induced functional differentiation. We also report for the first time that UDCA can induce senescence in colon cancer cells as assessed by flattened, spread out and vacuolated morphology as well as by senescence marker beta-galactosidase staining. We also found that UDCA inhibits the telomerase activity. Surprisingly, we found that UDCA is not a histone deacytylase inhibitor but instead induces hypoacetylation of histones unlike hyperacetylation induced by sodium butyrate. Our results also suggest that, although UDCA induced senescence is p53, p21 and Rb independent, HDAC6 appears to be important in UDCA induced senescence. In summary, our data shows that UDCA modulates chromatin by inducing histone hypoacetylation and induces differentiation and senescence in colon cancer cells.     

组蛋白乙酰化影响肿瘤细胞多药耐药性的研究进展

在临床治疗中,肿瘤细胞的多药耐药性（multi-drug resistance,MDR）已经成为化疗失败的重要原因之一。肿瘤细胞中组蛋白乙酰化异常与MDR的产生有关,组蛋白去乙酰化酶抑制剂（histone deacety-lases inhibitor,HDACi）能够抑制肿瘤生长,避免肿瘤产生耐药性,已经作为一种新型的抗肿瘤药物应用于临床。HDACi可能通过阻滞细胞周期、促进细胞分化、诱导细胞凋亡等多种生物学效应发挥其抗肿瘤的作用,HDACi与其他药物联合应用在抗肿瘤方面也展现了良好的应用前景。     

组蛋白乙酰化影响肿瘤细胞多药耐药性的研究进展

在临床治疗中,肿瘤细胞的多药耐药性(multi-drug resistance,MDR)已经成为化疗失败的重要原因之一。肿瘤细胞中组蛋白乙酰化异常与MDR的产生有关,组蛋白去乙酰化酶抑制剂(histone deacety-lases inhibitor,HDACi)能够抑制肿瘤生长,避免肿瘤产生耐药性,已经作为一种新型的抗肿瘤药物应用于临床。HDACi可能通过阻滞细胞周期、促进细胞分化、诱导细胞凋亡等多种生物学效应发挥其抗肿瘤的作用,HDACi与其他药物联合应用在抗肿瘤方面也展现了良好的应用前景。     

Subclinical doxorubicin-induced cardiotoxicity update: role of neutrophils and endothelium

Doxorubicin (DOX) is a highly effective chemotherapy agent that often causes cardiotoxicity. Despite a number of extensive studies, the risk for DOX cardiotoxicity remains unpredictable. The majority of the studies on DOX-induced cardiotoxicity have been focused on the effects on cardiomyocytes that lead to contractile dysfunction. The roles of systemic inflammation, endothelial injury and neutrophil recruitment, all induced by the DOX, are increasingly recognized as the mechanisms that trigger the development and progression of DOX-induced cardiomyopathy. This review explores recent data regarding the possible mechanisms and biomarkers of early subclinical DOX-associated cardiotoxicity.     

Chemoprevention of rat hepatocarcinogenesis with histone deacetylase inhibitors: Efficacy of tributyrin,a butyric acid prodrug

Hepatocellular carcinoma (HCC) ranks in prevalence and mortality among top 10 cancers worldwide. Butyric acid (BA), a member of histone deacetylase inhibitors (HDACi) has been proposed as an anticarcinogenic agent. However, its short half‐life is a therapeutical limitation. This problem could be circumvented with tributyrin (TB), a proposed BA prodrug. To investigate TB effectiveness for chemoprevention, rats were treated with the compound during initial phases of “resistant hepatocyte” model of hepatocarcinogenesis, and cellular and molecular parameters were evaluated. TB inhibited (p < 0.05) development of hepatic preneoplastic lesions (PNL) including persistent ones considered HCC progression sites. TB increased (p < 0.05) PNL remodeling, a process whereby they tend to disappear. TB did not inhibit cell proliferation in PNL, but induced (p < 0.05) apoptosis in remodeling ones. Compared to controls, rats treated with TB presented increased (p < 0.05) hepatic levels of BA indicating its effectiveness as a prodrug. Molecular mechanisms of TB‐induced hepatocarcinogenesis chemoprevention were investigated. TB increased (p < 0.05) hepatic nuclear histone H3K9 hyperacetylation specifically in PNL and p21 protein expression, which could be associated with inhibitory HDAC effects. Moreover, it reduced (p < 0.05) the frequency of persistent PNL with aberrant cytoplasmic p53 accumulation, an alteration associated with increased malignancy. Original data observed in our study support the effectiveness of TB as a prodrug of BA and as an HDACi in hepatocarcinogenesis chemoprevention. Besides histone acetylation and p21 restored expression, molecular mechanisms involved with TB anticarcinogenic actions could also be related to modulation of p53 pathways. © 2008 Wiley‐Liss, Inc.     

The influence of the time interval between monoHER and doxorubicin administration on the protection against doxorubicin-induced cardiotoxicity in mice

Purpose: Despite its well-known cardiotoxicity, the anthracyclin doxorubicin (DOX) continues to be an effective and widely used chemotherapeutic agent. DOX-induced cardiac damage presumably results from the formation of free radicals by DOX. Reactive oxygen species particularly affect the cardiac myocytes because these cells seem to have a relatively poor antioxidant defense system. The semisynthetic flavonoid monohydroxyethylrutoside (monoHER) showed cardioprotection against DOX-induced cardiotoxicity through its radical scavenging and iron chelating properties. Because of the relatively short final half-life of monoHER (about 30 min), it is expected that the time interval between monoHER and DOX might be of influence on the cardioprotective effect of monoHER. Therefore, the aim of the present study was to investigate this possible effect. Methods: Six groups of 6 BALB/c mice were treated with saline, DOX alone or DOX (4 mg/kg i.v.) preceded by monoHER (500 mg/kg i.p.) with an interval of 10, 30, 60 or 120 min. After a 6-week treatment period and additional observation for 2 weeks, the mice were sacrificed. Their cardiac tissues were processed for light microscopy, after which cardiomyocyte damage was evaluated according to Billingham (in Cancer Treat Rep 62(6):865–872, 1978). Microscopic evaluation revealed that treatment with DOX alone induced significant cardiac damage in comparison to the saline control group (P<0.001). Results: The number of damaged cardiomyocytes was 9.6-fold (95% CI 4.4–21.0) higher in mice treated with DOX alone than that in animals of the control group. The ratio of aberrant cardiomyocytes in mice treated with DOX preceded by monoHER and those in mice treated with saline ranged from 1.6 to 2.8 (mean 2.2, 95% CI 1.2–4.1, P=0.019). The mean protective effect by adding monoHER before DOX led to a significant 4.4-fold reduction (P<0.001, 95% CI 2.3–8.2) of abnormal cardiomyocytes. This protective effect did not depend on the time interval between monoHER and DOX administration (P=0.345). Conclusion: The results indicate that in an outpatient clinical setting monoHER may be administered shortly before DOX.This work was supported in part by grant VU-97-1525 from the Koningin Wilhelmina Foundation, Amsterdam, The Netherlands.     

Anti-tumour activity in vitro and in vivo of selective differentiating agents containing hydroxamate.

A series of hydroxamates, which are not metalloprotease inhibitors, have been found to be selectively toxic to a range of transformed and human tumour cells without killing normal cells (fibroblasts, melanocytes) at the same concentrations. Within 24 h of treatment, drug action is characterized by morphological reversion of tumour cells to a more normal phenotype (dendritic morphology), and rapid and reversible acetylation of histone H4 in both tumour and normal cells. Two hydroxamates inhibited growth of xenografts of human melanoma cells in nude mice; resistance did not develop in vivo or in vitro. A third hydroxamate, trichostatin A, was active in vitro but became inactivated and had no anti-tumour activity in vivo. Development of dendritic morphology was found to be dependent upon phosphatase activity, RNA and protein synthesis. Proliferating hybrid clones of sensitive and resistant cells remained sensitive to ABHA, indicating a dominant-negative mechanism of sensitivity. Histone H4 hyperacetylation suggests that these agents act at the chromatin level. This work may lead to new drugs that are potent, and selective anti-tumour agents with low toxicity to normal cells.     

Effects of amifostine on perfused isolated rat heart and on acute doxorubicin-induced cardiotoxicity

Purpose: To determine the effects of amifostine on an isolated perfused rat-heart model and its protective activity with regard to cardiotoxic doxorubicin perfusion. Methods: Langendorff constant-pressure isolated rat-heart preparations were used to analyze the effects of the drugs during a 40-min period of perfusion after a 20-min stabilization interval. The first study was conducted with amifostine alone (controls and 10⁻⁶, 10⁻⁵, and 10⁻⁴ M amifostine; n=6 in each group). The second study was conducted with amifostine and doxorubicin (controls, 2.5 × 10⁻⁵ M doxorubicin, 2.5 × 10⁻⁵ M doxorubicin and 10⁻⁵ M amifostine, and 2.5 × 10⁻⁵ M doxorubicin and 10⁻⁴ M amifostine; n=4 in each group). Results: Amifostine had no significant effect on hemodynamic parameters at 10⁻⁶, 10⁻⁵, and 10⁻⁴ M concentrations. However, amifostine increased the coronary flow expressed as a percentage ± SEM of the baseline flow as follows: 82 ± 4% for controls, 95 ± 6% for 10⁻⁶ M amifostine, (P=0.13), 111 ± 4% for 10⁻⁵ M amifostine (P < 0.01), and 104 ± 3% for 10⁻⁶ M amifostine (P < 0.01). When we commenced an amifostine perfusion 20 min in advance of and then during a 40-min perfusion with doxorubicin, at a cardiotoxic concentration of 2.5 × 10⁻⁵ M the left ventricular pressures (LVDP, expressed as percentages ± SEM of the baseline LVDP before doxorubicin) were 55 ± 3% for the doxorubicin controls, 68 ± 2% for doxorubicin with 10⁻⁵ M amifostine (P=0.05), and 80 ± 3% for doxorubicin with 10⁻⁴ M amifostine (P < 0.01). Whether this protective effect might be related to the known free-radical-scavenging activity of amifostine remains to be determined. Conclusion: On a Langendorff-type model of rat heart, 10⁻⁵ and 10⁻⁴ M amifostine alone induced a coronary dilation and, when associated with a cardiotoxic concentration of 2.5 × 10⁻⁵ M doxorubicin, 10⁻⁵ and 10⁻⁴ M amifostine displayed a cardioprotective effect. Received: 9 March 1998 / Accepted: 6 July 1998     

Inactivation of class II transactivator by DNA methylation and histone deacetylation associated with absence of HLA-DR induction by interferon-gamma in haematopoietic tumour cells

By presenting immunogenic peptides at the cell surface, major histocompatibility complex (MHC) class II molecules play a key role in the control of adaptive immune responses. Whether expressed constitutively or induced by interferon-gamma, expression of MHC class II molecules is regulated via coactivator class II transactivator (CIITA); moreover, suppression of their expression is one mechanism by which cancer cells escape host immunity. In this study, we surveyed the relationship between the expression of one MHC class II antigen, HLA-DR, and its coactivators in a group of haematopoietic cell lines, and explored the role of the aberrant DNA methylation in silencing HLA-DR expression. Among 26 cell lines studied, HLA-DR expression was lost from eight T-cell and two myeloid leukaemia cell lines, and this loss was closely associated with suppression of CIITA-PIV expression. Notably, nine of the 10 cell lines that lost CIITA-PIV expression showed methylation of the gene's 5' CpG island. Thus, DNA methylation is believed to inhibit the expression of MHC class II molecules in haematopoietic tumour cells by silencing its coactivator, CIITA-PIV. Furthermore, methylation of CIITA-PIV was detected in seven of 32 primary acute myeloid leukaemia specimens, indicating that epigenetic alteration is not a cell line-specific phenomenon. Collectively, these data suggest that, by suppressing expression of MHC class II molecules, epigenetic inactivation of CIITA provides a survival advantage to a subset of haematopoietic tumours.     

辛二酰苯胺异羟肟酸对结肠癌HCT116和SW480细胞增殖、周期和凋亡的影响

目的:研究辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)对结肠癌细胞系HCT116和SW480增殖、周期和凋亡的影响,并对其分子作用机制进行初步探讨.方法:将不同浓度SAHA分别处理结肠癌HCT116和SW480细胞后,MTT法检测SAHA对HCT116和SW480细胞增殖的影响,流式细胞仪检测HCT116和SW480细胞周期和细胞凋亡率,罗丹明(rhodamine) 123和二氯二氢荧光素二乙酸酯(DCFH-DA)法检测HCT116和SW480细胞线粒体跨膜电位(Aψm)和活性氧(ROS)水平,Real-time PCR和Western blotting法检测乙酰化组蛋白3(Ac-H3)、p21、CyclinD1、Bax和Bcl-2的mRNA和蛋白的表达水平.结果:SAHA作用于HCT116和SW480细胞48 h后,细胞增殖被抑制、细胞周期G1期比率升高、凋亡率升高(均P＜0.05),线粒体跨膜电位显著下降、细胞内ROS产生增多(均P＜0.05).与对照组比较,SAHA处理组p21和Bax mRNA增多、Cyclin D1和Bcl-2 mRNA表达量减少(均P＜0.05),相关蛋白Ac-H3、p21和Bax增多,CyclinD1和Bcl-2减少(均P＜0.05).结论:SAHA抑制结肠癌HCT116和SW480细胞增殖、阻滞细胞周期并诱导细胞凋亡,其机可能与调节p21、CyclinD1和Bcl-2家族基因的表达、促进组蛋白乙酰化有关.     

Aberrant methylation and histone deacetylation of cyclooxygenase 2 in gastric cancer.

Cyclooxygenase 2 plays a critical role in the development of gastrointestinal cancers in both human and animal models. About 80% of the gastric cancer showed a high level of expression of cyclooxygenase 2, but a subset of cases do not express without unknown reason. Aberrant methylation of CpG island of COX-2 was examined by using a series of gastric cancer cell lines and primary gastric cancers. Two out of 8 cell lines (25%) and 11 out of 93 (12%) primary cancers showed aberrant methylation of the 5' region of COX-2. Methylation of COX-2 was closely associated with loss of expression and treatment of methylation inhibitor, 5-deoxy-2'-azacytidine restored the expression of COX-2. A combined treatment of 5-deoxy-2'-azacytidine and a histone deacetylese inhibitor, trichostatin A, restored re-expression of the gene synergistically and chromatin immunoprecipitation analysis revealed that histone of methylated COX-2 promoter is deacetylated, indicating the role of cytosine methylation and histone deacetylation in the silencing of the gene. These results indicate that a subset of gastric cancer with COX-2 methylation evolves through the pathway that is independent of COX-2 expression and that COX-2 inhibitor may not be useful to induce apoptosis in these cases.     

Possible role of formaldehyde in the apoptotic and mitotic effect of 1-methyl-ascorbigen

PC-3 human prostate carcinoma cells were treated with 100 ug/ml 1-methyl-ascorbigen (Me-Asc). This treatment resulted in a significant decrease in tumor cell number in parallel with an increase in apoptotic cells. The formaldehyde (HCHO) level in the culture medium was also increased. Dimedone (Di), a known capture molecule forming formaldemethone with HCHO, applied simultaneously with Me-Asc in 10 ug/ml doses diminished the apoptosis-inducing effect of Me-Asc. The possible role of in situ generated HCHO in the induction of apoptosis is discussed.