Pure drug nano-assemblies: A facile carrier-free nanoplatform for efficient cancer therapy

Nanoparticulate drug delivery systems (Nano-DDSs) have emerged as possible solution to the obstacles of anticancer drug delivery. However, the clinical outcomes and translation are restricted by several drawbacks, such as low drug loading, premature drug leakage and carrier-related toxicity. Recently, pure drug nano-assemblies (PDNAs), fabricated by the self-assembly or co-assembly of pure drug molecules, have attracted considerable attention. Their facile and reproducible preparation technique helps to remove the bottleneck of nanomedicines including quality control, scale-up production and clinical translation. Acting as both carriers and cargos, the carrier-free PDNAs have an ultra-high or even 100% drug loading. In addition, combination therapies based on PDNAs could possibly address the most intractable problems in cancer treatment, such as tumor metastasis and drug resistance. In the present review, the latest development of PDNAs for cancer treatment is overviewed. First, PDNAs are classified according to the composition of drug molecules, and the assembly mechanisms are discussed. Furthermore, the co-delivery of PDNAs for combination therapies is summarized, with special focus on the improvement of therapeutic outcomes. Finally, future prospects and challenges of PDNAs for efficient cancer therapy are spotlighted.

Protective effect of exogenous biliverdin against ultraviolet B-induced photodamage in HaCaT cells / 中华皮肤科杂志

Objective To evaluate the protective effect of exogenous biliverdin on ultraviolet B (UVB)-radiated HaCaT cells,and to explore its mechanism.Methods Cultured HaCaT cells were divided into 5 groups:control group receiving no treatment,UVB group irradiated with 30 mJ/cm2 UVB,3 biliverdin + UVB groups treated with 100 nmol/L,1 μmol/L and 10 μmol/L biliverdin respectively for 1 hour followed by 30 mJ/cm2 UVB radiation.After 24-hour treatment,changes in the morphology of HaCaT cells were observed,and cell counting kit 8 (CCK8) assay was performed to determine cell survival rates in the above groups.Western blot analysis was conducted to measure the protein expression of antioxidant signaling molecule NF-E2-related factor-2 (Nrf-2) and the photodamage signaling molecules matrix metalloproteinase-1 (MMP-1) and MMP-3.Results CCK8 assay showed that the survival rate of HaCaT cells was significantly lower in the UVB group than in the control group (P ＜ 0.05),but significantly higher in the 100-nmol/L,1-μmol/L and 10-μmol/L biliverdin + UVB groups than in the UVB group (all P ＜ 0.05).Western blot analysis showed that the protein expression of MMP-1 and MMP-3 was significantly higher in the UVB group (1.150 ± 0.187,0.979 ± 0.054 respectively) than in the control group (0.116 ± 0.018,0.636 ± 0.035 respectively;both P ＜ 0.01),but was significantly lower in the 100-nmol/L,1-μmol/L and 10-μmol/L biliverdin + UVB groups (MMP-1:0.825 ± 0.139,0.313 ± 0.047 and 0.286 ± 0.036 respectively;MMP-3:0.888 ± 0.017,0.672 ± 0.042 and 0.569:±:0.037 respectively) than in the UVB group (all P ＜ 0.05).Moreover,the protein expression of Nrf-2 was significantly lower in the UVB group (0.906 ± 0.034) than in the control group (1.242 ± 0.141,P ＜ 0.05),but significantly higher in the 100-nmol/L,1-μmol/L and 10-μmol/L biliverdin + UVB groups (1.556 ± 0.112,1.897 ± 0.234 and 2.035 ±0.274) than in the UVB group (all P ＜ 0.01).Conclusion Exogenous biliverdin protects against UVB-induced photodamage in HaCaT cells,which may be associated with Nrf-2 antioxidant signaling pathway.

Association of biliverdin reductase A gene polymorphisms with neonatal hyperbilirubinemia from Fujian area / 中华实用儿科临床杂志

Objective To assess the association of single nucleotide polymorphisms (SNPs)of biliverdin reductase A (BLVRA) with neonatal hyperbilirubinemia from Fujian area.Methods A total of 286 patients with neonatal hyperbilirubinemia and 250 healthy controls were enrolled.Genotypes of 5 SNPs within BLVRA gene including rs699512,rs1802846,rs7738,rs1637530 and rs2302032 were determined with matrix-assisted laser desorption ionization/time of flight mass spectrometer.The frequencies of genotype,allele,haplotype and their differentiations were analyzed.Results All 5 SNPs had conformed to Hardy-Weinberg equilibrium (all P ＞ 0.05).rs699512 and rs1637530 showed a significant difference between the 2 groups in both allelic and genotypic frequencies (all P ＜ 0.05),but no significant differences were found in the other SNPs(all P ＞ O.05).In recessive model,the frequency of rs699512 GG genotype of patients was significantly lower than that of the healthy control group(OR =0.494,95％ CI:0.276-0.886,P =0.018),while in dominant model,the frequencies of rs699512 GG + AG and rs1637530 TT + CT genotype of patients were significantly lower than that of the healthy control group(OR =0.678,0.627;95％ CI:0.482-0.954,0.444-0.885;P =0.026,0.008).Based on linkage disequilibrium analysis and haplotype construction,rs1637530,rs2302032,rs699512 and rs1802846 locus in the same area.Based on haplotype CGAT,TGGT,CTAT and CGGT had significant differences between the 2 groups (all P ＜ 0.05),and could reduce the risk of high blood bilirubin (OR =0.588,0.687,0.501;95％ CI:0.434-0.797,0.496-0.952,0.250-1.004).Conclusions rs699512 and rs1637530 may be associated with neonatal hyperbilirubinemia,A allele in rs699512 and C allele in rs1637530 may be associated with significantly increased risk of neonatal hyperbilirubinemia.

Biliverdin protects against cisplatin-induced apoptosis of renal tubular epithelial cells / 华中科技大学学报（医学）（英德文版）

Biliverdin (BV) has long been thought to be a cytotoxic metabolic waste product. It has also been demonstrated to have important cytoprotective functions during oxidative stress. The present study aimed to examine the cytoprotective effect of BV on NRK-52E cells, a proximal tubular cell line derived from rat kidney. Cells were treated with 50 µmol/L cisplatin for 24 h (cisplatin group) or pre-treated with BV for 30 min, then with 50 µmol/L cisplatin for 24 h (cisplatin+BV group). Those given no treatment served as a control. Cell apoptosis was evaluated by flow cytometry and cell viability by Cell Counting Kit-8 (CCK-8). The protein expressions of cleaved caspase3, Bax and Bcl-2 were assessed by Western blotting. Reactive oxygen species (ROS) levels were measured using carboxydichlorodihydrofluorescein diacetate (H2DCF). The results showed that cisplatin induced the apoptosis of NRK-52E cells, decreased cell viability, and increased the formation of ROS by upregulating the expression of cleaved caspase3 and Bax and decreasing Bcl-2 protein expression. These effects could be significantly reversed by pretreatment with BV. It was concluded that BV can protect against cisplatin-induced cell apoptosis through the anti-oxidative effects.

Análise e identificação de produtos do catabolismo de heme nas formas epimastigotas de Trypanosoma cruzi / Analysis and identification of heme catabolism products in Trypanosoma cruzi epimastigotes forms

O Trypanosoma cruzi, agente etiológico da doença de Chagas, possui um ciclo de vida complexo, deve lidar com diversas condições do ambiente e depende dos hospedeiros para suprir suas necessidades nutricionais. Uma delas é a necessidade de captar a molécula de heme (Fe-protoporfirina IX) que será utilizada como fator de crescimento. Os mecanismos envolvendo o metabolismo de heme são cruciais para a sobrevivência do T. cruzi pois o parasito não possui várias enzimas de biossíntese dessa porfirina e o heme livre pode apresentar citotoxicidade para célula. Na tentativa de perseguir o destino final do heme no parasito, nós estudamos essa via inexplorada no T. cruzi. Nessa tese, nós demonstramos que epimastigotas cultivados com heme, produziram os compostos, α-meso hidroxiheme, verdoheme e biliverdina (identificados por HPLC acoplado á espectrofotômetria). Além disso, nós observamos através de análise dos extratos de epimastigotas no espectrômetro de massas (LQT Orbitrap), espécies iônicas de m/z 583,4 e m/z 619,3. A fragmentação subsequente desses íons originaram espécies filhas típicas das moléculas de biliverdina e verdoheme, respectivamente. Nós observamos também, espécies iônicas de m/z 1397,4 e m/z 1135,4. A fragmentação dessas espécies produziram íons, sendo um deles com a mesma massa molecular de heme (m/z 616,3). Essa espécie iônica por sua vez, gerou fragmentos iônicos idênticos a uma molécula de heme, confirmando que esses intermediários são produtos da modificação da porfirina. Baseado nesses resultados, nós propomos um modelo onde o catabolismo de heme em T. cruzi, envolveria a conjugação da bis(glutationil)spermina, um derivado da tripanotiona presente em tripanossomatídeos, à porfirina (m/z 1137,4), seguido da remoção de dois resíduos de ácidos glutâmicos (m/z 1135,4)...

Trypanosoma cruzi, the causal agent of Chagas disease, has a complex life cycle and they must cope with diverse environmental conditions and depends on hosts for its nutritional needs. One of the nutritional characteristic is that they need a heme compound (Fe-protoporphyrin IX) as a growth factor. The mechanisms involved in these processes are crucial for their survival mainly because of trypanosomatids lack of the complete heme biosynthetic pathway and the cytotoxic activity of free heme. Following the fate of this porphyrin in the parasite we studied this missing pathway in T. cruzi. Here, we show that epimastigotes cultivated with heme yielded the compounds, α-meso hydroxyheme, verdoheme and biliverdin (as determined by HPLC with diode array detector). Furthermore, we observed ion species of m/z 583.4 and m/z 619.3 from epimastigotes extracts detected by direct infusion on LQT Orbitrap platform. A tipical biliverdin and verdoheme doughter-ion species were generated by m/z 583.4 and m/z 619.3 fragmentations, respectively. We also observed an ion species at m/z 1397.4 and m/z 1135. The subsequent fragmentation of this species produced a daughter-ions whose one with the same molecular mass as heme (m/z 616.4). This species, in turn, generated daughter species identical to an authentic heme, confirming that these intermediates were modified heme products. Based on these findings, we propose that heme catabolism in T. cruzi involves a additions of Bis(glutathionyl)spermine, a low molecular mass thiols occurring in trypanosomatids, to heme (m/z 1397.4), followed by removal of the glutamic residues (m/z 1135)...

Biliverdin reductase A in the prevention of cellular senescence against oxidative stress

Biliverdin reductase A (BLVRA), an enzyme that converts biliverdin to bilirubin, has recently emerged as a key regulator of the cellular redox cycle. However, the role of BLVRA in the aging process remains unclear. To study the role of BLVRA in the aging process, we compared the stress responses of young and senescent human diploid fibroblasts (HDFs) to the reactive oxygen species (ROS) inducer, hydrogen peroxide (H2O2). H2O2 markedly induced BLVRA activity in young HDFs, but not in senescent HDFs. Additionally, depletion of BLVRA reduced the H2O2-dependent induction of heme oxygenase-1 (HO-1) in young HDFs, but not in senescent cells, suggesting an aging-dependent differential modulation of responses to oxidative stress. The role of BLVRA in the regulation of cellular senescence was confirmed when lentiviral RNAitransfected stable primary HDFs with reduced BLVRA expression showed upregulation of the CDK inhibitor family members p16, p53, and p21, followed by cell cycle arrest in G0-G1 phase with high expression of senescence-associated beta-galactosidase. Taken together, these data support the notion that BLVRA contributes significantly to modulation of the aging process by adjusting the cellular oxidative status.

Heme Oxygenase-1: Its Therapeutic Roles in Inflammatory Diseases

Heme oxygenase (HO)-1 is an inducible enzyme that catalyzes the first and rate-limiting step in the oxidative degradation of free heme into ferrous iron, carbon monoxide (CO), and biliverdin (BV), the latter being subsequently converted into bilirubin (BR). HO-1, once expressed during inflammation, forms high concentrations of its enzymatic by-products that can influence various biological events, and this expression is proven to be associated with the resolution of inflammation. The degradation of heme by HO-1 itself, the signaling actions of CO, the antioxidant properties of BV/BR, and the sequestration of ferrous iron by ferritin all concertedly contribute to the anti-inflammatory effects of HO-1. This review focuses on the anti-inflammatory mechanisms of HO-1 actions and its roles in inflammatory diseases.

Heme Oxygenase-1: Its Therapeutic Roles in Inflammatory Diseases

Heme oxygenase (HO)-1 is an inducible enzyme that catalyzes the first and rate-limiting step in the oxidative degradation of free heme into ferrous iron, carbon monoxide (CO), and biliverdin (BV), the latter being subsequently converted into bilirubin (BR). HO-1, once expressed during inflammation, forms high concentrations of its enzymatic by-products that can influence various biological events, and this expression is proven to be associated with the resolution of inflammation. The degradation of heme by HO-1 itself, the signaling actions of CO, the antioxidant properties of BV/BR, and the sequestration of ferrous iron by ferritin all concertedly contribute to the anti-inflammatory effects of HO-1. This review focuses on the anti-inflammatory mechanisms of HO-1 actions and its roles in inflammatory diseases.

Detection of Biliverdin Reductase Activity of Schistosoma japonicum / 中国寄生虫学与寄生虫病杂志

Objective To explore the presence of biliverdin reductase (BR) activity in adult worms of Schistosoma japonicum in vitro. Methods The soluble fraction was isolated from homogenates of adult worms of S. japonicum, and incubated with biliverdin under different pH conditions and buffers. The time dependency of this enzymatic reaction was also detected. ResultsThe soluble fraction of the homogenates of adult worms could degrade biliverdin in vitro, the BR activity was 43.30 nmol/(mg?min), with its optimal condition at pH 8.7. The rate of reaction peaked at 15 min of incubation for the BR activity. Conclusion The presence of BR activity in adult worms of S. japonicum was firstly demonstrated.

Effects of echinacoside on protein expression from substantia nigra and striatal tissue in mouse MPTP model of Parkinsons disease by using 2-dimensional electrophoresis analysis / 中国药理学通报

Aim To study the effect of echinacoside on behavior and proteins expression from substantia nigra and striatal tissue in MPTP mouse model of Parkinsons disease(PD)and discover the mechanism of its potential dopaminergic neuroprotective effect in the protein level.Methods The mouse model of PD was induced by 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)and the behavioral analysis of C57BL/6 mice was performed by using spontaneous movement and rotarod test.A proteomic approach based on 2-dimensional electrophoresis(2-DE),mass spectrometry(MS)and figure analysis was used to evaluate the effect of echinacoside on the behavior and the protein expression in substantia nigra and striatal tissue in C57BL/6 mice after MPTP administration.Results ① Compared with control,MPTP lesion significantly reduced the number of spontaneous movement and latent period of mice on the rotating rod(both P