OBJECTIVETo evaluate the effect of lanthanum chloride on expressions of collagen protein and find a way to prevent and treat scar.

METHODSFour linear incisions were made on the dorsal skin of an adult, female Sprague-Dawley rat as an animal model. One was non-manipulated as a control; the second was injected with distilled water as a sham-control; the third was injected with 50 mmol/L of lanthanum chloride, and the fourth was injected with 50 micrograms neutralizing antibody of TGF-beta 1 as a positive control. All of the wound tissues were harvested and assayed with ABC method in 14 days and 28 days after the surgery.

RESULTSThe expressions of type I, III and IV of collagen protein in the third group significantly reduced in 14 and 28 days after the operation, compared with the control or sham-control group. Its values wen as similar as the fourth group.

CONCLUSIONLanthanum chloride could inhibit the expressions of collagen protein, and it may be used to prevent and treat scars.

Animals ; Collagen ; biosynthesis ; drug effects ; Female ; Lanthanum ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Wounds and Injuries ; metabolism

OBJECTIVETo investigate the effects of lanthanum chloride on the apoptosis of fibroblasts in trauma tissue.

METHODSFifty adult female SD rats were used and linear incisions were made on the back near the joints of extremities of the rats. One of the cuts receiving no treatment was designated as blank control (C). 0.25 ml of distilled water, lanthanum chloride (50 mmol/L) and the antibody of (TGFbeta(1)) transforming growth factor beta(1) (0.2 mg/ml) were respectively injected into the both sides of the other three wounds subcutaneously and the wounds were divided into simulating control (SC), lanthanum chloride (LC) and antibody (A) groups. The fibroblast apoptosis in the wound tissue samples and the change in intracellular calcium concentration (Ca(2+)) were determined by flow cytometry (FCM) and TUNEL methods on the 14th and 28th day after the injection.

RESULTSApoptosis of fibroblasts was enhanced significantly after 14 days of injection in LC and A groups compared with that in C and SC groups (P < 0.05 approximately 0.01). Furthermore, intracellular Ca(2+) was increased evidently in LC group (P < 0.01).

CONCLUSIONIt is indicated that lanthanum chloride might be effective in preventing scar development.

Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cicatrix ; prevention & control ; Female ; Fibroblasts ; drug effects ; pathology ; Flow Cytometry ; Lanthanum ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Wound Healing ; drug effects

Objective: To investigate the effect of nuclear factor erythroid 2-related factor 2 (Nrf2) in the alteration of tight junction protein expression in choroid plexus epithelial cells created by lanthanum-activated matrix metalloproteinase 9 (MMP9) . Methods: In October 2020, immortalized rat choroid plexus epithelial cell line (Z310) cells were used as the blood-cerebrospinal fluid barrier in vitro, and were divided into control group and 0.125, 0.25, 0.5 mmol/L lanthanum chloride (LaCl(3)) treatment group. After treating Z310 cells with different concentrations of LaCl(3) for 24 hours, the morphological changes of Z310 cells were observed under inverted microscope, the protein expression levels of MMP9, occludin and zonula occludens-1 (ZO-1) were observed by cellular immunofluorescence method, and the protein expression levels of MMP9, tissue inhibitors of metalloproteinase1 (TIMP1) , occludin, ZO-1 and Nrf2 were detected by Western blotting. The level of reactive oxygen species (ROS) in cells was detected by flow cytometry. Results: Compared with the control group, Z310 cells in the LaCl(3) treatment group were smaller in size, with fewer intercellular junctions, and more dead cells and cell fragments. The expression level of MMP9 protein in cells treated with 0.25 and 0.5 mmol/L LaCl(3) was significantly higher than that in the control group (P<0.05) , and the expression level of TIMP1 and tight junction proteins occudin and ZO-1 was significantly lower than that in the control group (P<0.05) . Compared with the control group, the ROS production level in the 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly increased (P<0.05) , and the Nrf2 protein expression level in the 0.125, 0.25, 0.5 mmol/L LaCl(3) treatment group was significantly decreased (P<0.05) . Conclusion: Lanthanum may increase the level of ROS in cells by down regulating the expression of Nrf2, thus activating MMP9 to reduce the expression level of intercellular tight junction proteins occludin and ZO-1.
Rats ; Animals ; Matrix Metalloproteinase 9/metabolism* ; NF-E2-Related Factor 2/metabolism* ; Tight Junction Proteins/metabolism* ; Occludin/pharmacology* ; Choroid Plexus/metabolism* ; Reactive Oxygen Species/metabolism* ; Lanthanum/pharmacology* ; Epithelial Cells ; Zonula Occludens-1 Protein/metabolism* ; Phosphoproteins/pharmacology*

OBJECTIVETo investigate the regulatory effects of lanthanum chloride (LaCl3) on the activation of nuclear factor kappa B inhibitor (IκB) kinase beta (IKKβ) induced by tumor necrosis factor alpha (TNF-α).

METHODS(1) Hela cells were cultured routinely in vitro. One portion of cells were collected and divided into TNF-α group (cultured with serum-free RMPI 1640 medium containing 20 ng/mL TNF-α for 30 min), low-dose LaCl3 + TNF-α group, moderate-dose LaCl3 + TNF-α group, high-dose LaCl3 + TNF-α group, LaCl3 group (cultured with serum-free RMPI 1640 medium containing 100 µmol/L LaCl3 for 30 min), and control group (cultured with serum-free RMPI 1640 medium for 30 min) according to the random number table. Cells in low-dose LaCl3 + TNF-α group, moderate-dose LaCl3 + TNF-α group, high-dose LaCl3 + TNF-α group were first cultured with serum-free RMPI 1640 medium containing 5, 25, 100 µmol/L LaCl3 for 4 h, and then stimulated with serum-free RMPI 1640 medium containing 20 ng/mL TNF-α for 30 min. There were 3 samples in each group. Cells were collected for detection of intracellular location of NF-κB/p65 protein by immunofluorescence staining. (2) Another portion of cells were collected and divided into TNF-α group, low-dose LaCl3 + TNF-α group, moderate-dose LaCl3 + TNF-α group, high-dose LaCl3 + TNF-α group, and control group with the same treatment as above. There were 3 samples in each group. The protein levels of NF-κB/p65 in nuclei, and the protein levels of IκBα, phosphorylated IκBα (p-IκBα) as well as IKKβ and phosphorylated IKKβ (p-IKKβ) in cytoplasm were determined by Western blotting. The binding activity between NF-κB/p65 in the nuclear and target gene was determined by NF-κB/p65 transcription factor kit (denoted as absorption value). Data were processed with analysis of variance or LSD-t test.

RESULTS(1) High expression of NF-κB/p65 was observed in cytoplasm of control group. High expression of NF-κB/p65 was observed in nuclei of TNF-α group. The expression of NF-κB/p65 in cytoplasm of LaCl3 group was lower than that of control group. In groups treated with LaCl3 and TNF-α, NF-κB/p65 expression levels in nuclei and cytoplasm were decreased along with the increase in the concentration of LaCl3, which were all lower than those in TNF-α group. (2) There was certain amount of NF-κB/p65 protein expressed in nuclei of control group. The expression of NF-κB/p65 protein in nuclei of TNF-α group was higher than that of control group. In groups treated with LaCl3 and TNF-α, the expressions of NF-κB/p65 protein in nuclei were decreased along with an increase in the concentration of LaCl3. The level of IκBα in TNF-α group was significantly decreased but that of p-IκBα increased as compared with those in control group. Along with the increase in the concentration of LaCl3, the levels of IκBα gradually increased and the levels of p-IκBα gradually decreased in groups treated with LaCl3 and TNF-α. There were no statistical differences in expression levels of IKKβ among the 5 groups. The expression of p-IKKβ could be hardly observed in control group, but it was obviously increased in TNF-α group. The expression levels of p-IKKβ in groups treated with LaCl3 and TNF-α were gradually decreased along with the increase in the concentration of LaCl3. The absorption value in TNF-α group was 0.39 ± 0.03, which was higher than that in control group (0, t = -7.23, P<0.01). The absorption values in low-dose LaCl3 +TNF-α group, moderate-dose LaCl3 + TNF-α group, and high-dose LaCl3 +TNF-α group were respectively 0.17 ± 0.03, 0.15 ± 0.03, and 0, which were obviously lower than that in TNF-α group (with t values respectively -6.54, -5.92, -7.23, P values all below 0.01).

CONCLUSIONSLaCl3 can block the activation of NF-κB signaling pathway by blocking the phosphorylation of IKKβ of Hela cells.

Culture Media ; HeLa Cells ; Humans ; I-kappa B Kinase ; metabolism ; I-kappa B Proteins ; metabolism ; Lanthanum ; pharmacology ; NF-KappaB Inhibitor alpha ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study influence of lanthanum chloride (LaCl(3)) on the expression of immediate early genes (IEGs) including c-jun, early growth response gene 1 (Egr1) and activity-regulated cytoskeletal gene (Arc) in the hippocampus of rats, and discuss the mechanism of LaCl(3) undermining learning and memory capability.

METHODSForty female Wistar adult rats were divided into control group, low LaCl(3)-contaminated group (0.25%), medium LaCl(3)-contaminated group (0.50%), and high LaCl(3)-contaminated group (1.00%) by randomized design. Each group had ten female rats along with five male rats and mated by the ratio of 2:1. The amounts of pups in the above four groups were 80, 83, 78 and 75 separately. The pups in respective group were La-dyed by lactation, and then the pups in LaCl(3)-contaminated groups drank 0.25%, 0.50% and 1.00% LaCl(3) separately for one month. Learning and memory capability of pups were measured in jumping stairs experiment. Hippocampal lanthanum content was determined by inductively coupled plasma mass spectrometry (ICP-MS). Hippocampal c-jun, Egr1 and Arc mRNA expression was detected by RT-PCR, and corresponding protein expression was measured by Western blotting method.

RESULTSIn the jumping stairs experiment, pups in 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups respectively made (1.75 ± 0.71), (2.38 ± 0.92) and (3.00 ± 0.76) mistakes; significantly higher than control group (1.25 ± 0.46) (q values were 4.386, 6.793, P < 0.05). However, the incubation period of 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups were (174.13 ± 33.72), (139.25 ± 45.83) and (75.50 ± 18.56) respectively, which were all significantly lower than that of control group (206.75 ± 20.47) (q values were 2.958, 6.121, 11.902, P < 0.05). Hippocampal c-jun mRNA expression were (0.89 ± 0.08), (0.77 ± 0.12), (0.58 ± 0.14) and (0.29 ± 0.10); while the c-jun protein expression were (0.72 ± 0.13), (0.64 ± 0.11), (0.43 ± 0.11) and (0.31 ± 0.14), and the Egr1 mRNA expression were (0.78 ± 0.09), (0.61 ± 0.13), (0.53 ± 0.10) and (0.22 ± 0.08), Egr1 protein expression were (0.65 ± 0.18), (0.40 ± 0.15), (0.32 ± 0.13) and (0.14 ± 0.09) in 0.25%, 0.50% and 1.00% LaCl(3)-contaminated groups; and all of which presented a dose-effect relationship that the correlation coefficients of these parameters with dose were -0.900 (t = 11.309, P = 0.000), -0.969 (t = 7.058, P = 0.000), -0.898 (t = 11.179, P = 0.000) and -0.962 (t = 6.739, P = 0.000).

CONCLUSIONLaCl(3) undermines the learning and memory capability of rats, which is possibly related to lower expression of c-jun and Egr1 gene and protein induced by lanthanum in hippocampus.

Animals ; Early Growth Response Protein 1 ; metabolism ; Female ; Gene Expression ; Genes, Immediate-Early ; drug effects ; genetics ; Hippocampus ; drug effects ; metabolism ; Lanthanum ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the influence of lanthanum on lipopolysaccharide (LPS) induced NF-KB activation in murine peritoneal macrophage.

METHODSPeritoneal macrophages were isolated and cultured by routine method, and randomly divided into 5 groups: i. e, control group, LPS group (with LPS stimulation for 30 min), La3+ group (with 2.5 micromol/L La3+ group for 30 min) , La3+ + LPS group( with 1 microg/ml LPS stimulation for 30 min after 30 min incubation with DMEM-F12 containing 2.5 microM of lanthanum.) ; La3+/LPS group (with 2.5 microM of lanthanum stimulation for 30 min, and then with 1 microg/ml of LPS for another 30 min after lanthanum was removed. The location of NF-kappaB p65 subunit (NF-kappaB/p65) in Mphi was detected by immunofluorescence and fluorescence microscope. The binding activity of NF-kappaB/p65 with DNA in nuclei was detected by TransAMTM NF-kappaB/p65 Transcription Factor assay kit. Meanwhile, the expression of NF-kappaB/p65 in nuclei, as well as IkappaBalpha in cytoplasm was measured by Western blotting. TNF-alpha content in culture supernatant were detected by ELISA.

RESULTS(1) The green fluorescence in control, La3+, La3+ LPS and La+/LPS groups was mainly located in cytoplasm, while that in LPS group was located in nuclei. The fluorescent intensity in LPS group was (116 +/- 14), which was obviously higher than that in other 4 groups (42 +/-7,73 +/-30,48 +/- 11 and 67 +/- 19, respectively, P <0.01). (2) The IkappaBalpha protein level in cytoplasm in control (0.048 +/- 0.027), La3+ group (0.062 +/- 0.049), La3+ + LPS group (0.066 +/-0.031) and La3+/LPS group (0.108 +/- 0.017) was significantly lower than that in LPS group (0.435 +/-0.066, P <0.01). (3) The expression and activation of nucleus p65 protein in Mphi in LPS group was obviously higher than the other 4 groups, but changes in the IkappaBalpha expression between LPS group and other 4 groups was of controversy. (4) TNFalpha level in the culture supernatant in La3+ group was lower than that in control group ( P < 0.05) and below the detection limit (25 pg/ml). Moreover, it in La3+ + LPS group and La3*/LPS group was lower than that in LPS group (P <0.01), but higher than that in control group.

CONCLUSIONLPS can activate the nucleus translocation of NF-kappaB/p65 in Mphi of mice, increase NF-KB/p65 expression and activity, but reduce IkappaBalpha protein expression, which lead to increase of TNFalpha secretion. Lanthanum can inhibit lipopolysaccharide induced NF-kappaB activation.

Animals ; Cells, Cultured ; I-kappa B Proteins ; metabolism ; Lanthanum ; pharmacology ; Lipopolysaccharides ; Macrophages, Peritoneal ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; NF-KappaB Inhibitor alpha ; Random Allocation ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore the influence of lanthanum chloride (LaCl3) on inducible nitric oxide synthase (iNOS) expression in RAW264.7 macrophages with lipopolysaccharide (LPS) induction, and to investigate its possible mechanisms.

METHODSThe RAW264.7 macrophages were randomly divided into four groups: i. e, control group (without treatment), LaCl3 group (with treatment of 2.5 micromol/L of LaCl3 for 24 hrs), LaCl3 + LPS group (with treatment of 2.5 micromol/L LaCl3 for 24h), and LPS group (with treatment of 1 mg/L LPS for 24 hrs). The iNOS protein expression was measured by immunofluorescence and Western blot. iNOS gene expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR). NO production in culture supernatant was assayed by nitrate reductase method.

RESULTSImmunofluorescence analysis showed that iNOS was located mainly in the cytoplasm. RAW264.7 cells with overexpression of iNOS accounted for 44.4%, which was obviously higher than that in LaCl3 + LPS group (11.8%, P < 0.05). There was a faint signal of FITC-labeled green tint in control group or LaCl3 group. The iNOS mRNA and protein expression, and the NO content in LPS group were significantly higher than those in control, LaCl3, and LaCl3 + LPS groups (P < 0.05).

CONCLUSIONLaCl3 can suppress LPS-induced iNOS overexpression at mRNA and protein level and reduce NO production, indicating that LaCl3 can antagonize the excessive activation of iNOS induced by LPS.

Animals ; Cell Line ; Lanthanum ; pharmacology ; Lipopolysaccharides ; toxicity ; Macrophages ; drug effects ; metabolism ; Mice ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the influence of lanthanum chloride on the TNFalpha expression of murine peritoneal macrophages stimulated by lipopolysaccharide (LPS).

METHODSMurine peritoneal macrophages (Mphi) were isolated, cultured and then stimulated by LPS. The influence of lanthanum chloride on the TNFalpha secretion and TNFalphamRNA expression of murine Mphi stimulated by LPS was determined by ELISA method and SYBR green fluorescence quantitative RT-PCR. Forty BALB/C mice were randomly divided into two groups and were treated by lethal dose of LPS and lanthanum chloride processed LPS, respectively. The mortality within 7 days was observed.

RESULTSThe TNFalpha secretion and TNFalphamRNA expression level of the Mphi from mice treated by lanthanum chloride processed LPS were obviously lower than those by LPS only (P < 0.01). The mortality of the mice treated by lethal dose of LPS which has been processed by lanthanum chloride was significantly lower than that by lethal dose of LPS only.

CONCLUSIONLanthanum chloride possessed the capacity of lowering down the toxicity of LPS and inhibiting the TNFalpha secretion and TNFalphamRNA expression in murine Mphi stimulated by LPS.

Animals ; Cells, Cultured ; Female ; Gene Expression Regulation ; drug effects ; Lanthanum ; pharmacology ; Lipopolysaccharides ; pharmacology ; toxicity ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics ; secretion

AIMTo study the biochemistry of lanthanides, the cooperative action of inorganic and organic anti-tumor drugs.

METHODSA series of rare earth complexes were synthesized with Ln(NO3) 6H2O, Phen and 5-Fu. Their anti-tumor activity was measured by the improved MTT, SRB methods.

RESULTSThe formula of complex Ln[(Phen)2(5-Fu)3(NO3)](NO3)2(Ln = Y, La, Ce, Sm, Gd, Dy, Er; Phen = 1, 10-phenanthroline; 5-Fu = fluorouracil) was characterized by elemental analyses, molar conductivity, IR, TGA, and 13C NMR spectra. The preliminary biological activity studies indicated that Lanthanide complex has strong anti-tumor activity in vitro.

CONCLUSIONThe complex might have anti-tumor cooperation action.

Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cerium ; chemistry ; Drug Synergism ; Dysprosium ; chemistry ; Erbium ; chemistry ; Fluorouracil ; chemistry ; Gadolinium ; chemistry ; Humans ; Lanthanoid Series Elements ; chemistry ; Lanthanum ; chemistry ; Phenanthrolines ; chemistry ; Samarium ; chemistry ; Structure-Activity Relationship ; Yttrium ; chemistry

OBJECTIVETo explore the preventive effect of lanthanum chloride on enteral bacterial translocation in scalded rats.

METHODSNinety Sprague-Dawley (SD) rats were employed in the study and randomly divided into three groups, i.e. normal control (A), burn control (B) and treatment (C) groups. Plasmid PUC19 labelled by JM109 was transfected to Escherichia coli (E. coli), so that restriction endonuclease finger - print image spectrum analysis could be applied to the tracing and quantification of the translocation of E. coli from intestine to mesenteric lymph nodes (MLNs) and blood. The intestinal tissue contents of endotoxin (ET), nitric oxide (NO), nitric oxide synthase (NOS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined.

RESULTSIt was identified that the bacteria in MLNs and blood exhibited the same gene map with those from gastric gavage in B and C groups. But the bacterial quantity in MLNs in C group on 3 postburn day (PBD) was much lower than that in B group (P < 0.05). The intestinal MDA content in C group on 1 and 3 PBDs was obviously higher than that in B group (P < 0.05).

CONCLUSIONBacteria (E. coli) could be translocated from gut to MLNs and blood, which could be evidently alleviated by lanthanum chloride by means of its bactericidal property, inhibition of NOS activity, so that NO production decreased, and its ability to increase SOD activity leading to less production of MDA.

Animals ; Burns ; drug therapy ; microbiology ; Endotoxins ; blood ; metabolism ; Escherichia coli ; drug effects ; growth & development ; metabolism ; Escherichia coli Infections ; blood ; microbiology ; prevention & control ; Female ; Intestines ; drug effects ; metabolism ; microbiology ; Lanthanum ; pharmacology ; Lymph Nodes ; drug effects ; microbiology ; Male ; Malondialdehyde ; blood ; metabolism ; Mesentery ; drug effects ; microbiology ; Nitric Oxide ; blood ; metabolism ; Nitric Oxide Synthase ; blood ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; metabolism