Selective inhibition of inflammatory cytokine storm in macrophages by α-momorcharin / 中国药理学通报

Aim To observe the inhibitory effect of Alpha-momorcharin (α-MMC) on the inflammatory cytokine storm of Ml-type inflammatory macrophages induced by LPS and explore its possible targeting mechanism. Methods Western blot was used to detect the expression of WIL2-S B lymphocytes, H9 T lymphocytes, THP-1 monocytes and M0 macrophages LRP1 receptor protein. CCK-8 method was used to detect the survival rate of the four cells. ELISA was used to detect the expression level of inflammatory cytokines in Ml macrophages. Western blot was used to detect the expression of TLR4 signaling pathway-related protein in Ml macrophages. Results Macrophages had a high density of LRP1 receptors consistent with monocytes; the survival rate of α-MMC on the four cells was positively correlated with the density of this receptor; α-MMC inhibited the expression of inflammatory cytokinesTNF-α, IL-lβ, IL-6, IL-8, MlP-lα and MCP-1 in Ml macrophages in a dose-and time-dependent manner; α-MMC showed significant inhibition to TAKl/pTAK1, p-JNK, p-APl and p-p65 signaling proteins of the TLR4 signaling pathway, and this inhibition could be blocked by the LRP1 receptor blocker RAP. Conclusions α-MMC selectively inhibits macrophage inflammatory cytokine synthesis by inhibiting TAK1 of the TLR4 signaling pathway, which in turn inhibits the downstream NF-ΚB and MAPK pathways, mediated by the LRP1 receptor. The selective immunosuppressive effect of α-MMC on macrophages may make it a very promising agent for the treatment of acute infectious macrophage activation syndrome (MAS).

Effects of tanshinone B regulating LRP6/Wntl/p-catenin pathway on cognitive function of mice with vascular dementia / 中国药理学通报

Aim To explore the protective mechanism of tanshinone B on cognitive dysfunction in mice with vascular dementia.Methods C57BL/6 male mice were divided into control group, vascular dementia model group(VD group), tanshinone B group(TSB 2,4,8 mg·kg-1), donepezil hydrochloride group(1 mg·kg-1), according to the random number table method.The VD model was constructed by the coarctation of bilateral common carotid arteries in mice.Ten days after the successful modeling, the low, medium, and high-dose tanshinone B groups were intraperitoneally injected with tanshinone B, the positive control medicine group was intraperitoneally injected with donepezil hydrochloride, once a day, and the mice in control group and VD group were injected intraperitoneally with the same amount of normal saline for 20 d.Morris water maze test was used to detect the learning and memory ability of mice in each group; the cortex and hippocampus of each group were separated, and MDA, SOD and GSH-Px were determined by spectrophotometry; the pathological changes in the hippocampus of each group were observed by HE staining.The expression of p-LRP6, LRP6, Wnt1 and β-catenin protein in the hippocampus of each group of mice were detected by Western blot.Results Compared with control group, the ability of memory was reduced, the content MDA increased(P<0.01), SOD and GSH-Px activities decreased(P<0.01), and significant pathological damage in hippocampus, the expression of p-LRP6, Wnt1, and β-catenin protein was significantly reduced in VD group(P<0.01).Compared with VD group, the learning and memory abilities of the mice were improved, the content of MDA decreased(P<0.01), the activities of SOD and GSH-Px increased(P<0.01), and the pathological damage in hippocampus was significantly improved.The expression of Wnt1 and β-catenin protein increased significantly in TSB treatment group(P<0.01).Conclusions TSB can improve the cognitive dysfunction of VD mice, and its mechanism may be related to the activation of the LRP6/Wnt1/β-catenin pathway in hippocampus.

Liuwei Dihuangwan Regulate Brain Microvessels in SAMP8 Mice Through RAGE/LRP1 Receptors / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Liuwei Dihuangwan on neurovascular injury in SAMP8 mice. MethodThe Alzheimer's disease （AD） model with insufficiency of kidney essence was induced in 75 SAMP8 mice aging 6 months. The model mice were divided into model group， positive control group （donepezil hydrochloride， 0.747 mg·kg-1·d-1）， and high-， medium-， and low-dose Liuwei Dihuangwan groups （2.700， 1.350， 0.675 g·kg-1·d-1）， with 15 mice in each group. Fifteen SAMR1 mice were assigned to a normal control group. All mice were administered continuously for 2 months. The spatial memory of mice was tested by the Morris water maze. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the hippocampus and cortex of brain tissues. The immunohistochemical method （IHC） was used to detect the deposition of amyloid β-protein （Aβ） and the expression of von Willebrand factor （vWF） and CD34 in the hippocampus and cortex of brain tissues. Electron microscopy was used to observe the ultrastructural changes in cerebral microvessels. Western blot was used to detect the protein expression levels of the receptor of advanced glycation endproduct （RAGE）， low-density lipoprotein receptor-related protein 1 （LRP1）， vascular endothelial growth factor A （VEGF-A）， and P-selection in the hippocampus and cortex of brain tissues. ResultCompared with the normal control group， the model group showed prolonged escape latency and swimming distance （P<0.01）， increased number of glial cells， decreased number of nerve cells， blurred tight junctions or enlarged gap of the brain microvascular endothelial cells， severely injured membrane structure， swollen mitochondria of endothelial cells， ruptured membrane， massive dissolution in cristae， increased protein expression of Aβ and vWF in the hippocampus and cortex （P<0.01）， reduced protein expression of CD34 （P<0.05）， elevated protein expression of RAGE and P-selection in the cortex （P<0.01）， and decreased protein expression level of LRP1 and VEGF-A （P<0.01）. Compared with the model group， the Liuwei Dihuangwan groups showed shortened escape latency and swimming distance （P<0.05）， reduced number of glial cells in the cortex and hippocampus， increased number of microvessels in the cortex， clear double-layer membrane structure in tight junctions between the microvascular endothelial cells， increased number of mitochondria with intact membrane and recovered mitochondrial cristae， decreased protein expression of Aβ， vWF， RAGE， and P-selection in the hippocampus and cortex （P<0.05）， and increased protein expression of CD34， LRP1， and VEGF-A （P<0.05）. ConclusionLiuwei Dihuangwan can regulate Aβ metabolism through the RAGE/LRP1 receptor system and promote cerebral microvascular angiogenesis by inhibiting vWF expression and increasing VEGF-A and CD34， thereby improving cerebral microvascular injury in SAMP8 mice.

The disbalance of LRP1 and SIRPα by psychological stress dampens the clearance of tumor cells by macrophages

The relationship between chronic psychological stress and tumorigenesis has been well defined in epidemiological studies; however, the underlying mechanism remains underexplored. In this study, we discovered that impaired macrophage phagocytosis contributed to the psychological stress-evoked tumor susceptibility, and the stress hormone glucocorticoid (GC) was identified as a principal detrimental factor. Mechanistically, GC disturbed the balance of the "eat me" signal receptor (low-density lipoprotein receptor-related protein-1, LRP1) and the "don't eat me" signal receptor (signal regulatory protein alpha, SIRPα). Further analysis revealed that GC led to a direct, glucocorticoid receptor (GR)-dependent trans-repression of LRP1 expression, and the repressed LRP1, in turn, resulted in the elevated gene level of SIRPα by down-regulating miRNA-4695-3p. These data collectively demonstrate that stress induces the imbalance of the LRP1/SIRPα axis and entails the disturbance of tumor cell clearance by macrophages. Our findings provide the mechanistic insight into psychological stress-evoked tumor susceptibility and indicate that the balance of LRP1/SIRPα axis may serve as a potential therapeutic strategy for tumor treatment.

Expressão de Proteína-1 Relacionada a Receptor de Lipoproteína de Baixa Densidade (LRP1) em Monócito em Correlação com EIMC em Pacientes Mexicanos Hipertensos / Monocyte Low-Density Lipoprotein Receptor-Related Protein 1 (LRP1) Expression Correlates with cIMT in Mexican Hypertensive Patients

Resumo Fundamento A hipertensão arterial (HTA) representa um grande fator de risco de morbidade e mortalidade cardiovascular. Ainda não se sabe que mecanismos moleculares específicos estão associados ao desenvolvimento de hipertensão essencial. Objetivo Neste trabalho, analisamos a associação entre expressão mRNA de monócito LRP1, expressão de proteína LRP1, e espessura íntima-média de carótida (EIMC) de pacientes com hipertensão essencial. Métodos A expressão mRNA de monócito LRP1 e os níveis de proteína e EIMC foram quantificados em 200 indivíduos mexicanos, sendo 91 normotensos (NT) e 109 hipertensos (HT) A significância estatística foi definida em p < 0,05. Resultados O grupo de pacientes HT tinha EIMC maior altamente significativa em comparação com os pacientes NT (p = 0,002), e isso está relacionado ao aumento na expressão mRNA de LRP1 (6,54 versus. 2,87) (p = 0,002) e expressão de proteína LRP1 (17,83 versus 6,25), respectivamente (p = 0,001). Essas diferenças foram mantidas mesmo quando dividimos nossos grupos de estudo, levando em consideração apenas aqueles que apresentavam dislipidemia na expressão de mRNA (p = 0,041) e de proteínas (p < 0,001). Também se identificou que a indução de LRP1 mediada por LRP1 em monócitos em de maneira dependente de dose e tempo, com diferença significativa em NT versus HT (0,195 ± 0,09 versus 0,226 ± 0,12, p = 0,046). Conclusão Foi encontrado um aumento em EIMC em indivíduos com hipertensão, associada a expressões de proteína LRP1 e mRNA mais altas em monócitos, independente da presença de dislipidemia em pacientes HT. Esses resultados que a upregulation de LRP1 em monócitos de pacientes hipertensos mexicanos poderia estar envolvida na diminuição da EIMC. (Arq Bras Cardiol. 2021; 116(1):56-65)

Abstract Background Arterial hypertension (HTA) represents a major risk factor for cardiovascular morbidity and mortality. It is not yet known which specific molecular mechanisms are associated with the development of essential hypertension. Objective In this study, we analyzed the association between LRP1 monocyte mRNA expression, LRP1 protein expression, and carotid intima media thickness (cIMT) of patients with essential hypertension. Methods The LRP1 monocyte mRNA expression and protein levels and cIMT were quantified in 200 Mexican subjects, 91 normotensive (NT) and 109 hypertensive (HT). Statistical significance was defined as p < 0.05. Results HT patients group had highly significant greater cIMT as compared to NT patients (p=0.002) and this correlated with an increase in the expression of LRP1 mRNA expression (6.54 vs. 2.87) (p = 0.002) and LRP1 protein expression (17.83 vs. 6.25), respectively (p = 0.001). These differences were maintained even when we divided our study groups, taking into account only those who presented dyslipidemia in both, mRNA (p = 0.041) and proteins expression (p < 0.001). It was also found that Ang II mediated LRP1 induction on monocytes in a dose and time dependent manner with significant difference in NT vs. HT (0.195 ± 0.09 vs. 0.226 ± 0.12, p = 0.046). Conclusion An increase in cIMT was found in subjects with hypertension, associated with higher mRNA and LRP1 protein expressions in monocytes, irrespective of the presence of dyslipidemias in HT patients. These results suggest that LRP1 upregulation in monocytes from Mexican hypertensive patients could be involved in the increased cIMT. (Arq Bras Cardiol. 2021; 116(1):56-65)

Escape from abluminal LRP1-mediated clearance for boosted nanoparticle brain delivery and brain metastasis treatment

Breast cancer brain metastases (BCBMs) are one of the most difficult malignancies to treat due to the intracranial location and multifocal growth. Chemotherapy and molecular targeted therapy are extremely ineffective for BCBMs due to the inept brain accumulation because of the formidable blood‒brain barrier (BBB). Accumulation studies prove that low density lipoprotein receptor-related protein 1 (LRP1) is promising target for BBB transcytosis. However, as the primary clearance receptor for amyloid beta and tissue plasminogen activator, LRP1 at abluminal side of BBB can clear LRP1-targeting therapeutics. Matrix metalloproteinase-1 (MMP1) is highly enriched in metastatic niche to promote growth of BCBMs. Herein, it is reported that nanoparticles (NPs-K-s-A) tethered with MMP1-sensitive fusion peptide containing HER2-targeting K and LRP1-targeting angiopep-2 (A), can surmount the BBB and escape LRP1-mediated clearance in metastatic niche. NPs-K-s-A revealed infinitely superior brain accumulation to angiopep-2-decorated NPs-A in BCBMs bearing mice, while comparable brain accumulation in normal mice. The delivered doxorubicin and lapatinib synergistically inhibit BCBMs growth and prolongs survival of mice bearing BCBMs. Due to the efficient BBB penetration, special and remarkable clearance escape, and facilitated therapeutic outcome, the fusion peptide-based drug delivery strategy may serve as a potential approach for clinical management of BCBMs.

Relationship between LRP5 gene single nucleotide polymorphism and glucocorticoid-induced osteoporosis in children / 中华内分泌外科杂志

Objective:To investigate the relationship between the polymorphisms of two SNP loci (rs901823 and rs3736228) in the low density lipoprotein receptor-associated protein 5 (LRP5) gene and glucocorticoids-induced osteoporosis (GIOP) in children.Methods:87 children with GIOP who were treated in Beijing Aiyuhua Women’s and Children’s Hospital and Beijing Children’s Hospital affiliated to Capital Medical University from Jan. 2015 to Dec. 2020 were selected as the research objects, and 100 children with normal bone mass who were treated with corticosteroids in this hospital during the same period were enrolled as the control group. Capillary electrophoresis and fragment analysis (SNaPshot) technology were used to genotype SNP sites rs901823 (T>C) and rs3736228 (C>T) ; Quantitative real-time fluorescence quantitative polymerase chain reaction detection method was employed to determine the relative mRNA of LPR5 gene The amount of expression.Results:For the rs901823 locus of the LRP5 gene, the TT, TC, and CC genotype distribution differences between the GIOP group and the control group were statistically significant ( χ2=14.176, P=0.001) . Compared with the TT genotype, carriers of the TC and CC genotypes had a higher risk of GIOP, with OR values of 3.022 (1.189-6.387) and 5.483 (1.452-20.883) ; For academic significance, OR values were 3.412 (1.795-6.587) and 4.352 (1.215-15.982) . For the rs3736228 locus, the distribution of CC, CT, TT genotypes between the GIOP group and the control group was significantly different ( χ2=9.597, P=0.008) . Compared with CC carriers, CT genotype carriers had a significantly increased risk of GIOP, with an OR value of 5.125 (1.721-16.241) . The result of a dominant model was statistically significant, with an OR value of 4.165 (1.335-14.652) , while for TT there was no statistically significant difference between the carrier and the CC genotype ( P=0.512) , and the results of the recessive model also showed no significant statistical significance ( P=0.887) . There was a statistically significant difference in the frequency distribution of T and C alleles at rs901823 between the GIOP group and the control group ( χ2=17.298, P<0.001) , and the difference in the frequency distribution of C and T alleles at rs3736228 was also statistically significant ( χ2=9.356, P=0.002) . The relative expression level of LRP5 gene mRNA in children with GIOP was 1.34±0.26, which was significantly lower than the expression level of LRP5 gene mRNA in children in the control group of 3.06±0.42 ( t=8.248, P<0.001) . Among children with GIOP, the relative expression of LRP5 gene mRNA in patients with rs901823 locus TT, TC, and CC genotypes was statistically significant ( P<0.001) ; the differences in rs901823 locus CC, CT, TT genotype patients were significant. Pairwise comparison of the relative expression of LRP5 gene mRNA showed that there was no significant difference between the TT group and the CT group ( P>0.05) , but the expression of the CC group was significantly higher than that of the CT group and the TT group ( P<0.05) . Conclusion:The rs901823 and rs3736228 polymorphisms of LRP5 gene are correlated with the occurrence of GIOP and can be used as genetic markers for predicting GIOP in children.

Effect of Zuoguiwan on Wnt/β-catenin Signaling Pathway in Ovariectomized Osteoporosis Model Rats / 中国实验方剂学杂志

Objective:To observe the effect of Zuoguiwan on bone metabolism and Wnt/<italic>β</italic>-catenin signaling pathway in ovariectomized osteoporotic rats model， and to explore the molecular biological mechanism of Zuoguiwan in the prevention and treatment of osteoporosis. Method:The rat model of postmenopausal osteoporosis was established by bilateral ovariectomy， 60 female SD rats were randomly divided into sham operation group， model group， positive group （estradiol valerate tablet 0.05 mg·kg^-1·d^-1） and low， middle and high dose groups of Zuoguiwan （5.5，11，22 g·kg^-1·d^-1）.After successful establishment of the model in the 13^th week， intragastric administration （<italic>ig</italic>） was given once a day for a total of 12 weeks. After administration， the histomorphological changes of femur in rats were observed by hematoxylin-eosin （HE） staining， the bone mineral density （BMD） and bone mineral content（BMC） of femur were measured by dual energy X-ray apparatus， and the biomechanical properties of bone were measured by MTS Acumen3 biomechanical testing system. The contents of bone alkaline phosphatase （BALP）， bone glaprotein（BGP），estradiol （E₂） ，and tartrate-resistant acid phosphatase （TRAP）， type Ⅰ procollagen N-terminal propeptide （PINP） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein level of Wnt2，<italic>β</italic>-catenin，low density lipoprotein related receptor protein 5 （LRP5） and the phosphorylation level of glycogen synthase kinase-3<italic>β</italic>（GSK-3<italic>β</italic>） in rat tibia. Result:Compared with sham operation group， the maximum load and stiffness of BMD，BMC， in the model group decreased significantly（<italic>P</italic><0.01）， the contents of E₂ and PINP in serum decreased significantly（<italic>P</italic><0.01）， the content of BALP，BGP，TRAP increased significantly（<italic>P</italic><0.01）， the expression levels of Wnt2，p-GSK-3<italic>β </italic>Ser9，LRP5 and <italic>β</italic>-catenin protein in bone tissue decreased significantly（<italic>P</italic><0.01）， the trabecula of femur became thinner and thinner， the number of bone trabeculae decreased. Compared with model group， the maximum load and stiffness of BMD，BMC， in estradiol group and Zuoguiwan group were significantly increased （<italic>P</italic><0.05，<italic>P</italic><0.01）， the contents of serum E₂ and PINP were significantly increased （<italic>P</italic><0.05，<italic>P</italic><0.01）， the content of BALP，BGP，TRAP was significantly decreased （<italic>P</italic><0.01）， and the expression level of Wnt2，p-GSK-3<italic>β</italic> Ser9，LRP5， <italic>β</italic>-catenin protein in bone tissue was significantly increased （<italic>P</italic><0.05，<italic>P</italic><0.01） ， the trabeculae of femur became thicker， the number increased， the structure was basically clear. Conclusion:Zuoguiwan has a certain preventive and therapeutic effect on osteoporosis in ovariectomized rats， and its mechanism may be related to increasing the level of estrogen， activating Wnt/<italic>β</italic>-catenin signaling pathway， up-regulating the expression of Wnt2 and LRP5 protein， inhibiting the activity of GSK-3<italic>β</italic>， reducing the degradation of <italic>β</italic>-catenin， coordinating the dynamic coupling balance between bone formation and bone resorption， correcting the disorder of bone metabolism and improving bone morphology.

HLY78 Attenuates Neuronal Apoptosis via the LRP6/GSK3β/β-Catenin Signaling Pathway After Subarachnoid Hemorrhage in Rats / 神经科学通报·英文版

Neuronal apoptosis is one of the essential mechanisms of early brain injury after subarachnoid hemorrhage (SAH). Recently, HLY78 has been shown to inhibit apoptosis in tumor cells and embryonic cells caused by carbon ion radiation through activation of the Wnt/β-catenin pathway. This study was designed to explore the anti-apoptotic role of HLY78 in experimental SAH. The results demonstrated that HLY78 attenuated neuronal apoptosis and the neurological deficits after SAH through the activation of low-density lipoprotein receptor-related protein 6 (LRP6), which subsequently increased the level of phosphorylated glycogen synthesis kinase 3 beta (p-GSK3β) (Ser9), β-catenin, and Bcl-2, accompanied by a decrease of p-β-catenin, Bax, and cleaved caspase 3. An LRP6 small-interfering ribonucleic acid reversed the effects of HLY78. In conclusion, HLY78 attenuates neuronal apoptosis and improves neurological deficits through the LRP6/GSK3β/β-catenin signaling pathway after SAH in rats. HLY78 is a promising therapeutic agent to attenuate early brain injury after SAH.

HLY78 Attenuates Neuronal Apoptosis via the LRP6/GSK3β/β-Catenin Signaling Pathway After Subarachnoid Hemorrhage in Rats / 神经科学通报·英文版

Neuronal apoptosis is one of the essential mechanisms of early brain injury after subarachnoid hemorrhage (SAH). Recently, HLY78 has been shown to inhibit apoptosis in tumor cells and embryonic cells caused by carbon ion radiation through activation of the Wnt/β-catenin pathway. This study was designed to explore the anti-apoptotic role of HLY78 in experimental SAH. The results demonstrated that HLY78 attenuated neuronal apoptosis and the neurological deficits after SAH through the activation of low-density lipoprotein receptor-related protein 6 (LRP6), which subsequently increased the level of phosphorylated glycogen synthesis kinase 3 beta (p-GSK3β) (Ser9), β-catenin, and Bcl-2, accompanied by a decrease of p-β-catenin, Bax, and cleaved caspase 3. An LRP6 small-interfering ribonucleic acid reversed the effects of HLY78. In conclusion, HLY78 attenuates neuronal apoptosis and improves neurological deficits through the LRP6/GSK3β/β-catenin signaling pathway after SAH in rats. HLY78 is a promising therapeutic agent to attenuate early brain injury after SAH.

Localization of PQ-LRP protein in Microsporum canis / 中华皮肤科杂志

Objective To determine the location of PQ-LRP protein in Microsporum canis using the enhanced green fluorescent protein(EGFP)as a marker.Methods The total RNA was extracted from Microsporum canis,and reversely transcribed into cDNA.The PQ-LRP gene was amplified by PCR using the above cDNA as the template.The fusion gene of PQ-LRP gene and EGFP gene was linked to the plasmid pCAMBIA 1300.Microsporum canis was subjected to Agrobacterium tumefaciens-mediated transformation,in order to achieve the integrated expression of the fusion gene LRP-EGFP in Microsporum canis under the regulation by the fungal universal promoter Ptrpc and terminator Ttrpc.Laser-scanning confocal microscopy was conducted to determine the cellular localization of the fusion protein.Results The expression vector pCAMBIA-LRP-EGFP was successfully constructed,and the fusion gene LRP-EGFP was expressed integratedly in Microsporum canis.Laser-scanning confocal microscopy showed that fluorescence signals of LRP-EGFP were concentrated on the cell membrane of Microsporum canis,giving a granular or cluster-like appearance.Conclusion The infusion gene LRP-EGFP can be successfully expressed in Microsporum canis,and PQ-LRP protein is located on the cell membrane of Microsporum canis.

The Effect on LRP6 via Regulating Canonical Wnt/β-catenin Signaling in Doxorubicin-induced Cardiomyocyte Injury / 中山大学学报(医学科学版)

@#【Objective】To investigate the function of LRP6 and canonical Wnt/β-catenin signaling pathway in Doxorubicin-induced cardiomyopathy.【Methods】To establish the model of Dox cardiomyopathy in vitro and in vivo，H9C2 cells were treated with Dox（1 μmol/L）for 12 h and twelve SD rats were divided into two groups equally，and intraperitoneally injected with normal saline and Dox respectively. The changes of protein and mRNA levels were detected by western blot and qPCR. Cardiomyocytes were transfected with siRNA to knockdown LRP6. Mitochondrial membrane potential，nuclear condensation and matrix swelling were determined by Rhodamine 123 ，Hoechst and Mitotracker staining respectively. The apoptosis rate of cells was measured by flow cytometric analysis. 【Results】 The model of Dox cardiomyopathy was successfully established in vitro and in vivo. Dox downregulated the mRNA and protein levels of LRP6. Knockdown of LRP6 aggravated the cell apoptosis and mitochondrial damage induced by Dox. Both Dox and silencing LRP6 induced the downregulation of β - catenin ，and activation of β - catenin reversed the cardiomyocytes apoptosis caused by Dox. 【Conclusions】 Dox downregulated the expression of LRP6 and inhibited canonical Wnt/β- catenin signaling pathway，thus causing cardiomyocytes apoptosis and mitochondrial dysfunction.

The low-density lipoprotein receptor-related protein 5 (LRP5) 4037C>T polymorphism: candidate for susceptibility to type 1 diabetes mellitus

ABSTRACT Objective: The present study has investigated the association between low-density lipoprotein receptor-related protein 5 (LRP5) 4037C>T polymorphism and type 1 diabetes mellitus (T1DM) susceptibility in a Brazilian population. Subjects and methods: A total number of 134 T1DM patients and 180 normoglycemic individuals (NG) aged 6-20 years were studied. Glycated hemoglobin and glucose levels were determined. Genotyping of LRP5 4037C>T (rs3736228) was performed. Results: T1DM patients showed poor glycemic control. Genotypes in the codominant (CT: OR = 2.99 [CI 95%: 1.71-5.24], p < 0.001; TT: OR = 5.34 [CI 95%: 1.05-2702], p < 0.001), dominant (CT + TT: OR = 3.16 [CI 95%: 1.84-5.43], p < 0.001) and log-additive (OR = 2.78 [CI 95%: 1.70-4.52], p < 0.001) models, and LRP5 4037T allele (OR = 2.88, [CI 95%: 1.78-4.77], p < 0.001) were associated with an increased risk of developing T1DM. LRP5 4037CT and CT+TT carriers in T1DM group showed higher concentrations of serum glucose and glycated hemoglobin when compared with CC carriers. Conclusion: The LRP5 4037C>T may represent a candidate for T1DM susceptibility, as well as poor glycemic control.

Research of Xiaoyan Decoction on multidrug resistance of lung adenocarcinoma A549/DDP cell with cisplatin resistance in human gene / 中草药

Objective: To study the Xiaoyan Decoction serum influence the expression level of lung adenocarcinoma that cisplatin-resistent of A549/DDP cell multidrug resistance-associated protein 1 (MRP1), lung resistance-related protein (LRP), MRP mRNA, and LRP mRNA, to find out Xiaoyan Decoction's target to the chemotherapy resistance, and to lay foundation for the clinical of reversing lung cancer chemotherapy resistance. Methods: The A549 nude mice hypodermic tumor model was given an equal amount of saline, 2 mg/kg of cisplatin, low and high doses of Xiaoyan Decoction (20, 40 g/kg), and then the serum was gain. The cisplatin resistance of human lung adenocarcinoma A549/DDP cell line was chosen as acquired drug-resistance model, Xiaoyan Decoction on MDR of A549/DDP cell gene expression product of multidrug resistance-associated protein 1, lung resistance-related protein and its effect on mRNA expression level was detected by Western blotting and RT-PCR technique. Results: Compared with the control group, the expression of MRP1 and LRP protein decreased gradually (P<0.05) as the serum drug concentration of Xiaoyan Decoction increased. The level of LRP mRNA and MRP1 mRNA in low and high doses of Xiaoyan Decoction group also decreased, and much more obvious inhibition of gene expression was observed as the increasing of drug concentration. Conclusion: Different serum drug concentration of Xiaoyan Decoction can differently control the expression of MRP1, LRP protein, MRP1 mRNA, and LRP mRNA, with the increasing of serum drug concentration, the decreasing of expression level, namely positive-correlated to the dose, which indicates that Xiaoyan Decoction Reversal of the lung cancer drug resistance the might related to the MRP1 and LRP.

Effects of Abnormal Phlegmatic Munziq on Ability of Learning and Memory and Protein Expressions of Brain Tissue RAGE and LRP1 of APP/PS1 Transgenic Mice Model of AD / 中国中医药信息杂志

Objective To investigate the effects of Abnormal Phlegmatic Munziq on ability of learning and memory, and protein expressions of brain tissue RAGE and LRP1 of APP/PS1 transgenetic mice model of AD;To discuss its mechanism of action. Methods Three-month-old APP/PS1 transgenic mice were randomly divided into 5 groups: model control group, positive control group, Abnormal Phlegmatic Munziq high-, medium-, and low-dose groups, 18 mice in each group. Another 18 three-month-old C57BL/6J mice were chosen as normal control group. All administration groups received relevant medicine for successive 6 months. Then the changes in ability of learning and memory of mice were detected by Step-down test; protein expressions of LRP1 and RAGE were detected by immunohistochemistry and Western blot. Results Compared with the normal control group, the reaction time of learning grades and the mistake times increased, incubation of memory grades decreased and the mistake times increased in the model control group (P<0.01);Compared with the model control group, the reaction time of learning grades and the mistake times decreased, incubation of memory grades increased and the mistake times decreased in all administration groups (P<0.05, P<0.01). Immunohistochemistry and Western blot results showed that compared with normal control group, the LRP1 expression decreased and RAGE increased in the model control group (P<0.05);Compared with the model control group, the LRP1 expression decreased and RAGE increased in Abnormal Phlegmatic Munziq high-, medium-, and low-dose groups (P<0.05,P<0.01). Conclusion Abnormal Phlegmatic Munziq can improve ability of spatial learning and memory in APP/PS1 mice and regulate the expressions of RAGE and LRP1.

Research progress on association of LRP6 gene polymorphism and metabolic syndrome / 中国药理学通报

Metabolic syndrome is the fundamental factor in the pathogenesis of a variety of diseases, and it has not yet been fully understood due to its complicated mechanism. Multiple re-searches have implicated that Wnt/β-catenin signaling pathway may have a significant effect on the formation and development of metabolic syndrome. LRP6 is an important co-receptor of Wnt/β-catenin signaling pathway ,and there are some researches im-plicating the correlation between LRP6 and metabolic syndrome. The in-depth research on the gene polymorphism and its modula-tion mechanism can provide new ideas and directions for meta-bolic syndrome therapy.

Androgen Receptor-dependent Expression of Low-density Lipoprotein Receptor-related Protein 6 is Necessary for Prostate Cancer Cell Proliferation

Androgen receptor (AR) signaling is important for prostate cancer (PCa) cell proliferation. Here, we showed that proliferation of hormone-sensitive prostate cancer cells such as LNCaP was significantly enhanced by testosterone stimulation whereas hormone-insensitive prostate cancer cells such as PC3 and VCaP did not respond to testosterone stimulation. Blocking of AR using bicalutamide abolished testosterone-induced proliferation of LNCaP cells. In addition, knockdown of AR blocked testosterone-induced proliferation of LNCaP cells. Basal expression of low-density lipoprotein receptor-related protein 6 (LRP6) was elevated in VCaP cells whereas stimulation of testosterone did not affect the expression of LRP6. However, expression of LRP6 in LNCaP cells was increased by testosterone stimulation. In addition, knockdown of LRP6 abrogated testosterone-induced proliferation of LNCaP cells. Given these results, we suggest that androgen-dependent expression of LRP6 plays a crucial role in hormone-sensitive prostate cancer cell proliferation.

Traditional Chinese Medicine Regulation on Aβ Transporter in Blood-brain Barrier / 世界科学技术-中医药现代化

The main pathological change of Alzheimer’s disease (AD) was the formation of senile plaques (SP) induced by the abnormal accumulation ofβ-amyloid protein (Aβ). Blood-brain barrier (BBB) can regulate the Aβ metabolism in the brain through relevant transporter to complete the across BBB transport. This paper introduced two main transporters, which were the receptor for advanced glycation end products (RAGE) and the low density lipoprotein receptor-related protein-1 (LRP-1) in BBB for the elucidation of neuronal cell toxicity induced by inflammatory factors from Aβ accumulation within the brain. It further explored the regulation on two transport proteins, which were RAGE and LRP-1 in BBB for the reduction of abnormal accumulation of Aβ, relieving of inflammation in the brain, and protection of cerebral neurons by traditional Chinese medicine (TCM).

Relationship between Physico-chemical Parameters and Phylogenetics Study of Human Low Density Lipoprotein Receptor-Related Protein (LRP).

In this study, 11members of human low density lipoprotein receptor-related protein (LRP) sequences was retrieved from UniProtKB/ SWISS-PROT protein database and was analyzed for information about their structural, functional and phylogenetic features. This was achieved by using many established biocomputational tools which was available at their latest version. This study shows that LRP 12 and 3 are closely related with LRP8 being their nearest neighbor. In all, it was observed that there were very low possession of certain essential amino acid like glycine, proline and a very high aliphatic in all the LRP family. Considering the evolutionary history, functional domains, high aliphatic index, overall proportion of glycine and proline and the established role of one (LRP8) of this closely related LRP in diseases, it is thus predicted that the other closely related LRP3 and 12 molecules may be important candidate in investigating the aetiopathology of Myocardial infarction1diseases or other heart related disorder.

Regulation of microRNA-7-5p and LRP6 by Epstein-Barr Virus-Encoded RNAs in Burkitt's Lymphoma Cell Line Akata

Epstein-Barr virus (EBV)-encoded small non-coding RNAs (EBERs) are abundantly expressed in various EBV-associated malignancies, and play critical roles in cell proliferation, tumorigenesis, and apoptosis resistance. However, the mechanism how EBERs regulate cell function awaits further clarification. In this study, we investigated the effect of EBERs on the expression of cellular microRNA (miRNA) and mRNA expression. To test the effect of EBERs while unaffected by other EBV genes, we used EBERs-deleted recombinant EBV infected Burkitt's lymphoma cell line (Akata(+)EBERs(-)) as well as EBV-infected (Akata(+)) and EBV uninfected (Akata(-)) cell lines. They all have the same genetic backgrounds. First, 15 different cellular miRNAs which have reverse complementary sequences to EBERs and have reported targets were selected by bioinformatics analysis. When RT-PCR was carried out for the 16 miRNAs using RNAs from Akata(+), Akata(-), and Akata(+)EBERs(-) cells, hsa-miR-7-5p was the only one showing down-regulated expression in Akata(+) than in Akata(-) and Akata(+)EBERs(-) cells. Bioinformatics and mRNA microarray analyses for Akata(+), Akata(-), and Akata(+)EBERs(-) cell lines were then carried out to predict putative targets of hsa-miR-7-5p. Among the 6 predicted targets of hsa-miR-7-5p, only low density lipoprotein receptor-related protein 6 (LRP6) was up-regulated in EBERs-expressing cells when tested by RT-PCR and Western blot. However, luciferase reporter assay showed that the 3'-UTR of LRP6 was not directly targeted by hsa-miR-7-5p. Our data suggest that both hsa-miR-7-5p and LRP6 are regulated by EBERs in Akata cells, and these genes may partly mediate the tumorigenic function of EBERs in Burkitt's lymphoma.