Potentilla anserina polysaccharide alleviates cadmium-induced oxidative stress and apoptosis of H9c2 cells by regulating the MG53-mediated RISK pathway.

Oxidative stress plays a crucial role in cadmium (Cd)-induced myocardial injury. Mitsugumin 53 (MG53) and its mediated reperfusion injury salvage kinase (RISK) pathway have been demonstrated to be closely related to myocardial oxidative damage. Potentilla anserina L. polysaccharide (PAP) is a polysaccharide with antioxidant capacity, which exerts protective effect on Cd-induced damage. However, it remains unknown whether PAP can prevent and treat Cd-induced cardiomyocyte damages. The present study was desgined to explore the effect of PAP on Cd-induced damage in H9c2 cells based on MG53 and the mediated RISK pathway. For in vitro evaluation, cell viability and apoptosis rate were analyzed by CCK-8 assay and flow cytometry, respectively. Furthermore, oxidative stress was assessed by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining and using superoxide dismutase (SOD), catalase (CAT), and glutathione/oxidized glutathione (GSH/GSSG) kits. The mitochondrial function was measured by JC-10 staining and ATP detection assay. Western blot was performed to detect the expression of proteins related to MG53, the RISK pathway, and apoptosis. The results indicated that Cd increased the levels of reactive oxygen species (ROS) in H9c2 cells. Cd decreased the activities of SOD and CAT and the ratio of GSH/GSSG, resulting in decreases in cell viability and increases in apoptosis. Interestingly, PAP reversed Cd-induced oxidative stress and cell apoptosis. Meanwhile, Cd reduced the expression of MG53 in H9c2 cells and inhibited the RISK pathway, which was mediated by decreasing the ratio of p-AktSer473/Akt, p-GSK3ßSer9/GSK3ß and p-ERK1/2/ERK1/2. In addition, Cd impaired mitochondrial function, which involved a reduction in ATP content and mitochondrial membrane potential (MMP), and an increase in the ratio of Bax/Bcl-2, cytoplasmic cytochrome c/mitochondrial cytochrome c, and Cleaved-Caspase 3/Pro-Caspase 3. Importantly, PAP alleviated Cd-induced MG53 reduction, activated the RISK pathway, and reduced mitochondrial damage. Interestingly, knockdown of MG53 or inhibition of the RISK pathway attenuated the protective effect of PAP in Cd-induced H9c2 cells. In sum, PAP reduces Cd-induced damage in H9c2 cells, which is mediated by increasing MG53 expression and activating the RISK pathway.

Potentilla anserine L. polysaccharide protects against cadmium-induced neurotoxicity.

Cadmium is a toxic metal that can damage the brain and other organs. This study aimed to explore the protective effects of Potentilla anserine L. polysaccharide (PAP) against CdCl2-induced neurotoxicity in N2a and SH-SY5Y cells and in the cerebral cortex of BALB/c mice. In addition, we aimed to identify the potential mechanisms underlying these protective effects. Relative to CdCl2 treatment alone, pretreatment with PAP prevented the reduction in cell viability evoked by CdCl2, decreased rates of apoptosis, promoted calcium homeostasis, decreased ROS accumulation, increased mitochondrial membrane potential, inhibited cytochrome C and AIF release, and prevented the cleavage of caspase-3 and PARP. In addition, PAP significantly decreased the CdCl2-induced phosphorylation of CaMKII, Akt, and mTOR. In conclusion, PAP represents a potential therapeutic agent for the treatment of Cd-induced neurotoxicity, functioning in part via attenuating the activation of the mitochondrial apoptosis pathway and the Ca2+-CaMKII-dependent Akt/mTOR pathway.

Potentilla anserine L. polysaccharide inhibits cadmium-induced neurotoxicity by attenuating autophagy.

Cadmium (Cd), a heavy metal with cytotoxicity, can activate autophagy. This study aimed to explore the effects and mechanisms of Potentilla anserine L. polysaccharide (PAP) on autophagy in N2a cells, primary neurons, and the brain of BALB/c mice exposed to Cd. The CCK-8 assay results showed that the cell viability decreased and the number of acidic vesicular organelles, autophagic vacuoles, lysosomes, and dysfunctional mitochondria increased in the cytoplasm of Cd-exposed N2a cells and primary neurons, as revealed by acridine orange staining, monodansylcadaverine staining, and transmission electron microscopy. PAP mitigated Cd-induced neuronal death and characteristic changes in autophagy. The expression of LC3 IILC3 II, Bcl-2, p62, Beclin-1, and PI3K class III was examined by Western blot analysis. Furthermore, the PI3K inhibitor (LY294002 or 3-MA) and/or PAP reversed the Cd-induced upregulated expression of LC3 II, Beclin-1, and PI3K class III, with a synergy between PI3K inhibitor and PAP against Cd-induced autophagy. The findings suggested that PAP partially prevented Cd-induced autophagic cell death in neurons by inhibiting the PI3K class III/Beclin-1 signaling pathway in vitro and in vivo.

Protective effect of Potentilla anserina polysaccharide on cadmium-induced nephrotoxicity in vitro and in vivo.

The aim of this research was to investigate the antioxidant and anti-apoptotic activities of Potentilla anserina polysaccharide (PAP) on kidney damage induced by cadmium (Cd) in vitro and in vivo. PAP has been suggested to have anti-oxidation, anti-apoptosis, immunoregulation, antimicrobial, antitussive, and expectorant abilities. In this study, PAP was extracted and the major components of PAP were analyzed. It was shown that PAP pretreatment remarkably improved redox homeostasis, both in human embryonic kidney 293 (HEK293) cells and in BALB/c mice. Administration of PAP attenuated the mitochondrial dysfunction, degeneration, and fibrosis of kidney induced by Cd. Furthermore, PAP exhibited anti-apoptotic activity, which involved regulating both the mitochondria-mediated intrinsic apoptotic pathway and the death receptor-initiated extrinsic pathway. These results suggest that PAP is a potential therapeutic agent for Cd-induced nephrotoxicity.

A coumarin-based two-photon probe for hydrogen peroxide.

A new fluorescence probe was developed for hydrogen peroxide (H2O2) detection based on donor-excited photo induced electron transfer (D-PET) mechanism, together with the benzil as a quenching and recognizing moiety. The benzil could convert to benzoic anhydride via a Baeyer-Villiger type reaction in the presence of H2O2, followed by hydrolysis of benzoicanhydride to give benzoic acid, and the fluorophore released. The probe was synthesized by a 6-step procedure starting from 4-(diethylamino)salicylaldehyde. A density functional theory (DFT) calculation was performed to demonstrate that the benzil was a fluorescence quencher. The probe was evaluated in both one-photon and two-photon mode, and it exhibited high selectivity toward H2O2 over other reactive oxygen species and high sensitivity with a detection limit of 0.09 µM. Furthermore, the probe was successfully applied to cell imaging of intracellular H2O2 levels with one-photon microscopy and two-photon microscopy. The superior properties of the probe made it of great potential use in more chemical and biological researches.

A coumarin-derived fluorescent chemosensor for selectively detecting Cu²âº: synthesis, DFT calculations and cell imaging applications.

A novel coumarin-based fluorescent probe L ((4E)-4-((7-hydroxy-4-methyl-2-oxo-2H-chromen-8-yl) methyleneamino)-1,2-dihyydro-2,3-dimethyl-1-phenylpyrazol-5-one) has been developed as a simple and efficient chemosensor which exhibits a significant fluorescence reduction in the presence of metal cations. This sensor exhibits high selectivity and sensitivity toward Cu(2+) over other common cations. The mechanism for detecting copper was evaluated by time-dependent density functional theory (TD-DFT) calculations and the coordination mode was also confirmed by density functional theory (DFT) calculations. Furthermore, results of cell imaging in this study indicate that this new probe may be useful for detection and monitoring of Cu(2+) in biological applications.

Gelatin functionalized graphene oxide for mineralization of hydroxyapatite: biomimetic and in vitro evaluation.

We report a facile modification of graphene oxide (GO) by gelatin to mimic charged proteins present in the extracellular matrix during bone formation. The bioinspired surface of GO-gelatin (GO-Gel) composite was used for biomimetic mineralization of hydroxyapatite (HA). A detailed structural and morphological characterization of the mineralized composite was performed. Additionally, MC3T3-E1 cells were cultured on the GO-Gel surfaces to observe various cellular activities and HA mineralization. Higher cellular activities such as cell adhesion, cell proliferation, and alkaline phosphatase activity (ALP) were observed on the GO-Gel surface compared with the GO or glass surface. The increase of ALP confirms that the proposed GO-Gel promotes the osteogenic differentiation of MC3T3-E1 cells. Moreover, the evidence of mineralization evaluated by scanning electron microscopy (SEM) and alizarin red staining (ARS) corroborate the idea that a native osteoid matrix is ultimately deposited. All these data suggest that the GO-Gel hybrids will have great potential as osteogenesis promoting scaffolds for successful application in bone surgery.

Selective off-on fluorescent chemosensor for detection of Fe3+ ions in aqueous media.

A Fe(3+) chemosensor L1 was successfully synthesized with a quinoline moiety bound to rhodamine 6G hydrazide. The sensor L1 shows high selectivity and sensitivity to Fe(3+) in aqueous solution in the presence of other trace metal ions in organisms, abundant cellular cations and prevalent toxic metal ions in the environment. In addition, biological imaging and micro computed tomography (MCT) technology studies have demonstrated that L1 could act as a turn-on fluorescent chemosensor for Fe(3+) in living cells.

An 1,3,4-oxadiazole-based OFF-ON fluorescent chemosensor for Zn2+ in aqueous solution and imaging application in living cells.

A new 1,3,4-oxadiazole-based fluorescence chemosensor 1, N-(2-ethoxy-2-oxoethyl)-N-(5-(2-hydroxy-3,5-di-tert-butylphenyl)-[1,3,4]oxadiazol-2-yl)glycine ethyl ester, has been designed and synthesized. Its fluorescence properties and selectivity for various metal ions were investigated in detail. A prominent fluorescence enhancement only for Zn(2+) was found in aqueous acetonitrile solution and the response mechanism of 1 was analyzed by time-resolved fluorescence decay and DFT calculations. Furthermore, the fluorescence imaging of Zn(2+) in living cells was successfully applied.

A sensitive colorimetric and ratiometric fluorescent probe for mercury species in aqueous solution and living cells.

A highly sensitive and selective fluorescent probe for inorganic and organic mercury species displays colorimetric and ratiometric response in a buffer solution via mercury promoted cleavage reaction. The probe is demonstrated to detect CH(3)HgCl in living cells.

[Relationship between V617F mutation and 46/1 haplotype in JAK2 gene in patients with chronic myeloproliferative diseases and frequencies of 46/1 haplotype in different Chinese nationalities].

Somatic gene V617F mutation in JAK2 is a critical molecular and biological indicator to diagnosis of chronic myeloproliferative disease (MPD). This study was aimed to investigate the genetic background of V617F mutation in 46/1 gene haplotype in Chinese MPD patients, and the frequencies of 46/1 gene haplotype and V617F mutation in three nationalities of Chinese populations. Peripheral blood or bone marrow samples of 150 V617F mutation positive MPD patients, 123 V617F mutation negative MPD patients, 124 healthy Han individuals, 395 healthy Tibetan individuals and 315 healthy Yugu individuals were collected. The allele-specific multiplex PCR method was established, the presence or absence of V617F mutation, the presence or absence of 46/1 haplotype, and the relationship between V617F and 46/1 haplotype were easily identified by agarose gel image. The results showed that the V617F mutation located in the 46/1 haplotype of 88 cases (58.67) among 150 V617F-positive MPD cases. In 814 Chinese healthy individuals including Han, Tibetan, Yugu nationalities, the frequency of the 46/1 gene haplotype was 38.37 without difference in the frequency among different nationalities, and no V617F mutation was found in Chinese healthy populations, The frequency of the 46/1 gene haplotype was 43.09 in V617F mutation negative MPD patients and was 69.33 in V617F mutation positive MPD patients, the latter was obviously higher than former and than that in healthy Han individuals. In conclusion, a multiplex PCR method has been developed that is simple and useful to identify V617F mutation in JAK2 gene and its relationship to the 46/1 haplotype. In more than half of Chinese V617F-positive MPD patients, the V617F mutation locates in 46/1 haplotype in JAK2. The frequencies of 46/1 haplotype are statistically insignificant among Han, Tibetan and Yugu nationality populations.

Recognition of copper and hydrogen sulfide in vitro using a fluorescein derivative indicator.

We report the development of a fluorescein-based chemosensor (L1) for monitoring ions or micromolecules (H(2)S). Copper ions are known to be toxic at high concentrations and hydrogen sulfide induces various problems. Herein we develop a simple method for detecting Cu(II) and H(2)S with high selectivity and sensitivity. The chemosensor L1 displays on-off-on type fluorescence change with alternately added Cu(II) and H(2)S to the media along with reversible forming-separating of the complex. The potential biomedical relevance of the chemical mechanisms involved in the detection of L1 is described.

[A study on human tongue cancer cells' proliferation affected by Lactobacillus acidophilus].

OBJECTIVE: To study the effects of Lactobacillus acidophilus (L. acidophilus) on the proliferation and cell cycle distribution of human tongue cancer cells (Tca8113 cells). METHODS: In vitro cultivated human Tca8113 cells were treated by L. acidophilus supernatant, inactivated bacilli, cell free extracts and normal culture medium respectively, which were 1, 4, 16-fold(s) dilutelly, to investigate the proliferous effects of Tca8113 cells using of inverted microscope, cell counting, sulforhodamine B (SRB) and flow cytometry. The free radicals and Ca2+ in Tca8113 cells were also studied by confocal laser scanning microscope (CLSM). RESULTS: At the 48th hour after adding different L. acidophilus components, the Tca8113 cells changed in shape from the diamond-like, polygonal and slabs into the elongated form. In the condition of different times and different culture concentrations, the proliferation of Tca8113 cells was significantly inhibited by L. acidophilus components, which enhanced as the time prolonged and the concentrations of each L. acidophilus components increased according to the cell counting and the SRB experimental analysis. The cell proliferation index (CPI) was significantly reduced (P<0.01). The free radicals and Ca2+ in Tca8113 cells under the effect of each L. acidophilus components for 48 h indicated an obviously rising (P<0.01). CONCLUSION: L. acidophilus restrains the proliferation of Tca8113 cells, which might be due to the increase in quantity of free radicals and Ca2+ in Tca8113 cells, and might be resulted from the release of metabolic products of L. acidophilus.

Cu(2+)-selective fluorescent chemosensor based on coumarin and its application in bioimaging.

A new fluorescent sensor L1 based on coumarin was synthesized. It shows high sensitivity and selectivity toward Cu(2+) in aqueous solution. The complexation mode and corresponding quenching mechanism were elucidated by ESI-MS and DFT calculations. In addition, biological imaging studies have demonstrated that L1 can detect Cu(2+) in living cells.

A turn-on chemosensor for Hg2+ in aqueous media and its application in "MCT" imaging in living cells.

A turn-on chemosensor L1, which exhibits high selectivity and sensitivity toward Hg(2+) over other common metal ions in aqueous media under a physiological pH window via a 1:1 binding mode, had been synthesized and characterized. L1 provides good fluorescent imaging of Hg(2+) in living cells. Particularly, we adopted the "micro computed tomography (MCT)" technology, successfully demonstrating the method of Hg(2+) sensing by L1 in cell lines, also the cell permeability of L1 and its imaging position in the cells.

A rhodamine-based "turn-on" fluorescent chemodosimeter for Cu2+ and its application in living cell imaging.

A new fluorescent probe 1, N-(Rhodamine-6G)lactam-hydrazinecarbothioamide, was synthesized as a fluorescent and colorimetric chemodosimeter in aqueous solution for Cu(2+). Following Cu(2+)-promoted ring opening, redox and hydrolysis reactions, comparable amplifications of absorption and fluorescence signals were observed upon addition of Cu(2+); this suggests that chemodosimeter 1 effectively avoided the fluorescence quenching caused by the paramagnetic nature of Cu(2+). Importantly, 1 can selectively recognize Cu(2+) in aqueous media in the presence of other trace metal ions in organisms, abundant cellular cations and the prevalent toxic metal ions in the environment with high sensitivity (detection limit <3 ppb) and a rapid response time (<2 min). In addition, the biological imaging study has demonstrated that 1 can detect Cu(2+) in the living cells.

A new rhodamine-based chemosensor for Cu2+ and the study of its behaviour in living cells.

A new rhodamine-based chemosensor (L1) was synthesized, and it exhibits high sensitivity and selectivity for the copper cation over other commonly coexistent metal ions in aqueous solution. Upon the addition of Cu(2+), the spirolactam ring of L1 was opened and a 1 : 1 metal-ligand complex was formed. Fluorescent imaging of Cu(2+) in living cells is also successfully demonstrated.

[Erythrocyte immunologic function effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor].

OBJECTIVE: To research the erythrocyte immunoregulation effects of Patrinia scabra extracts by macroporous adsorptive resins on mice burdened transplanted tumor. METHODS: Extracts of Patrinia scabra Bunge were separated by macroporous adsorptive resins, ingredients were analysised. Mice burdened transplanted tumor were given extracted drugs. Life prolongation rate was observed, erythrocyte immunologic function and the CD35, CD44s contents of red blood cell were evaluated. RESULTS: Polysaccharide and saponin accounted for 8.4% and 48.4%. Extracts could porolong life expectancy of mice, improve erythrocyte immunolgic function and increase the CD35 and CD44s contents of red blood cell. CONCLUSION: Extracts of Patrinia scabra Bunge by macroporous adsorptive resins have erythrocyte immunoregulation effects on mice burdened transplanted tumor.

[Effects of fuzheng yiliu granules on apoptotic rate and mitochondrial membrane potential of hepatocellular carcinoma cell line H22 from mice].

OBJECTIVE: To evaluate the effects of Fuzheng Yiliu Granules (FZYLG) on apoptotic rate and mitochondrial membrane potential (Delta psi m) of hepatocellular carcinoma cell line H22 from mice. METHODS: Forty-eight mice inoculated with H22 cells were randomly divided into four groups: untreated group, cyclophosphamide-treated group, high-dose FZYLG-treated group and low-dose FZYLG-treated group. After 14 days of corresponding treatment, H22 cells in each group were stained with propidium iodide, and the apoptotic rates were detected by flow cytometry (FCM). The rhodamine 123 was used as a fluorescence probe to label the H22 cells, and the fluorescence intensities were observed with laser scanning confocal microscope. The fluorescence intensity of H22 cells indicated the Delta psi m of H22 cells. RESULTS: FZYLG could significantly increase the apoptotic rate while reduce the Delta psi m of H22 cells from mice as compared with those in the untreated group. CONCLUSION: The antitumor effects of FZYLR on H22 cells from mice are related to decreasing the Delta psi m and then inducing the apoptosis of the H22 cells.

Proanthocyanidin from grape seeds enhances anti-tumor effect of doxorubicin both in vitro and in vivo.

The purpose of this study was to investigate the synergistic anti-tumor effect of proanthocyanidin (PA) and doxorubicin (DOX) on K562, A549 and CNE cells in vitro and experimental transplantation Sarcoma 180 (S180) and Hepatoma 22 (H22) in vivo and to explore the mechanism of its action. PA 12.5 approximately 100 mg/l inhibited proliferation of K562, A549, and CNE cells in vitro in a time- and concentration-dependent manner as determined by the microculture tetrazolium (MTT) assay. A combination of PA 12.5, 25 mg/l with DOX 0.01 approximately 1 mg/l treatment synergistically inhibited proliferation of K562, A549, and CNE cells with decreased IC50 values. Under the confocal laser scanning microscope, intracellular DOX, Ca2+, and Mg2+ concentrations were greatly increased whereas pH value and mitochondrial membrane potential were markedly reduced in K562 cells after treatment with a combination of PA plus DOX. At the same time, K562 cells showed morphological changes of apoptosis following treatment with PA plus DOX, and the administration of PA 25 mg/l plus DOX 0.3 mg/l for 24 h resulted in a significant increase in the percentage of apoptosis by flow cytometry as compared with DOX 0.3 mg/l alone (p < 0.05). In vivo experiments showed that a combination of PA 200 mg/kg i.g. with DOX 2 mg/kg i.p. treatment displayed an inhibitory effect on the growth of transplantation tumor S180 and H22 in mice compared with the DOX only group (p < 0.01). Taken together, these results suggest that PA enhances the DOX-induced anti-tumor effect and its mechanism is attributed to the promotion of DOX-induced apoptosis through increasing intracellular DOX, Ca2+ and Mg2+ concentrations, and reducing pH value and mitochondrial membrane potential.