Research progress on selective tooth agenesis caused by LRP6 gene mutation / 口腔疾病防治

@#Selective tooth agenesis (STA) is an abnormal number of teeth due to genetic factors or the environment and is most commonly observed for permanent teeth. LRP6 is one of the common causative genes of STA and is inherited by an autosomal dominant mechanism, leading to non-syndrome tooth agenesis (NSTA) or syndrome tooth agenesis (STA). NSTA is only involved in tooth number and appearance abnormalities, whereas STA caused by LRP6 gene mutation results abnormal ear development, oral-facial clefting, sparse hair and hypohidrosis. In this paper, we review the phenotype and gene mutation traits of selective STA caused by LRP6 gene mutation identified in recent years and describe 38 patients with tooth agenesis from 24 mutation sites of LRP6 gene. We analyzed the percentage of missing teeth and found that the lateral incisor in the maxilla and the second premolar in the maxilla and mandible were most commonly lost, whereas all central incisors in the maxilla remained. LRP6 gene plays a major role in tooth development via the WNT/β-catenin signaling pathway, and LRP6 gene mutation can lead to a series of abnormal manifestations due to the disruption of the signaling pathway. The literature showed that LRP6 gene mutations occurred mostly at the E1 or E2 subdomain, meaning that STA due to the mutants extracellularly disturbed the WNT/β-catenin signaling pathway. However, mature treatments for selective congenital tooth loss are lacking.

Advances in diagnosis and treatment of familial hypercholesterolemia / 南方医科大学学报

Familial hypercholesterolemia (FH) is an autosomal dominant inherited disease caused by abnormal lipoprotein metabolism. Patients with FH have a significantly increased risk of coronary artery disease (CAD) due to long-term exposure to high levels of low-density lipoprotein (LDL). The diagnosis of FH relies heavily on gene detection, and examination of LDL receptor (LDLR) function is of great significance in its treatment. This review summarizes the current advances in the screening, diagnosis, and treatment of FH and functional analysis of LDLR gene mutations.

LOX-1 Regulation in Anti-atherosclerosis of Active Compounds of Herbal Medicine: Current Knowledge and the New Insight / 中国结合医学杂志

Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) have recently been identified to be closely related to the occurrence and development of atherosclerosis (AS). A growing body of evidence has suggested Chinese medicine takes unique advantages in preventing and treating AS. In this review, the related research progress of AS and LOX-1 has been summarized. And the anti-AS effects of 10 active components of herbal medicine through LOX-1 regulation have been further reviewed. As a potential biomarker and target for intervention in AS, LOX-1 targeted therapy might provide a promising and novel approach to atherosclerotic prevention and treatment.

The role and mechanism of MMP13 and LRP1 in abnormal autophagy function of articular chondrocytes in patients with Kashin-Beck disease / 中华地方病学杂志

Objective:To investigate the impact of matrix metalloproteinase 13 (MMP13) and low-density lipoprotein receptor-related protein 1 (LRP1) on autophagy of articular chondrocytes in patients with Kashin-Beck disease (KBD).Methods:Human articular cartilage samples obtained from 4 KBD patients and 4 control subjects were collected from Shaanxi Institute for Endemic Disease Prevention and Control, and the expression levels of MMP13 and LRP1 in cartilage tissue were determined using immunohistochemistry (IHC). Chondrocytes were extracted and cultured in vitro, the mRNA and protein expression levels of LRP1 and the autophagy related genes [Beclin 1 (BECN1), microtubule associated protein 1 light chain 3 (LC3)], cartilage injury related genes [MMP13, caspase-3 (CASP3)], chondrocyte differentiation related genes [collagen type Ⅱ alpha 1 chain (COL2A1), and SRY-box transcription factor 9 (SOX9)] were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot (WB), respectively. Chondrocytes from 3 KBD patients were extracted, and MMP13 gene silencing experiment was performed by RNA interference (RNAi) technology, the mRNA and protein expression levels of the above genes were detected by qRT-PCR and WB, respectively. In addition, the antagonist receptor associated protein (RAP) of LRP1 was used to block the LRP1 of human normal chondrocytes (C28/I2 cells), and qRT-PCR and WB were used to detect the mRNA and protein expression levels of LRP1, chondrocyte autophagy, differentiation and cartilage injury related genes, respectively. Results:The IHC results showed that the expression levels of MMP13 (1.67 ± 0.21, 0.59 ± 0.15, 0.51 ± 0.12) in the surface, middle, and deep layers of cartilage tissue of KBD patients were significantly higher than those of control subjects (0.25 ± 0.03, 0.26 ± 0.04, 0.06 ± 0.01), and the differences were statistically significant ( t = - 11.38, P < 0.001; t = - 3.82, - 6.26, P = 0.019, 0.003). The expression levels of LRP1 (0.10 ± 0.02, 0.03 ± 0.01, 0.17 ± 0.03) were significantly lower than those of control subjects (1.63 ± 0.40, 0.44 ± 0.12, 0.34 ± 0.08), and the differences were statistically significant ( t = 6.61, 5.61, 3.64, P = 0.003, 0.005, 0.022). The mRNA and protein expression levels of MMP13, CASP3, SOX9 in chondrocytes of KBD patients were significantly higher than those of control subjects, and the differences were statistically significant ( P < 0.05). The mRNA expression levels of LRP1, LC3, COL2A1 were significantly lower than those of control subjects, and the differences were statistically significant ( P < 0.05). After silencing the MMP13 gene in chondrocytes of KBD patients, there were no significant differences in the mRNA and protein expression levels of LRP1, BECN1, LC3, CASP3, COL2A1, and SOX9 ( P > 0.05). After blocking LRP1 with RAP, the protein expression levels of LRP1, BECN1, LC3, MMP13, COL2A1 and SOX9 in chondrocytes were significantly lower than those in control group ( P < 0.05). Conclusions:There is no direct correlation between MMP13 and abnormal autophagy of articular chondrocytes in KBD patients. After blocking LRP1, the expression of the autophagy related genes BECN1 and LC3 in chondrocytes is decreased.

Bioinformatics Analysis of LDLR Gene Mutation that Shed a New Light on Red Yeast Rice Monacolin K Treatment � Systematic Review

Background: Hypercholesteremia is the major cause of cardiovascular diseases. It results from elevated cholesterol levels in the blood. LDL cholesterol is removed from the circulation by using the LDL receptor. Red mold rice or red yeast rice is produced by fermentation of the Monascus Purpureus yeast on rice. Many researchers suggest that the active component in Red Yeast Rice (monacolin k) serves as a treatment for hypercholesteremic patients. Methods: By using NCBI databases, specifically GenBank to analyze DNA sequence and mRNA sequence of LDLR gene. GenBank file format was helpful to extract an accession number of the gene, number of amino acids, exons, and length of nucleotides. FASTA format was also useful to retrieve the nucleotide sequence and get the function of the protein. BLAST was used to compare the protein product of the LDLR gene between humans and pan paniscus (pygmy chimpanzee). Results: In accession number NC_000019, the number of amino acids in protein product is 44389 bp, and the number of exons found is 18. On the other hand, the gene is located in chromosome 19. The function of LDLR gene is to control the production of LDL receptor where the low-density lipoprotein particles attach to it and are taken into the cell ending up in the lysosome where the protein is degraded and cholesterol is made which will inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase that controls the production of cholesterol. Finally, many organisms have the same gene like dogs, cows, mice, rats, zebrafish, and frogs. Conclusion: Mutation in the LDLR gene causing high level of cholesterol in the blood especially LDL (Low-density Lipoprotein). Monacolin k that found in red yeast rice (RYR) is safe and natural alternative treatment for hypercholesteremic patients by lowering the cholesterol level in the blood.

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.

Time dependent expression profiling of PTK2B and its relationship with Aβ, Tau and LRP-1 in hippocampus and blood of APPswe/PS1dE9 double-transgenic mouse / 中国应用生理学杂志

Objective: To uncover the time-dependent expression pattern of ptk2b gene and ptk2b-encoded protein, protein tyrosine kinase 2 beta(PTK2B), in the brain tissues of transgenic animal models of Alzheimer's disease (AD) and its relationship with the levels of Aβ1-42, phosphorylation of Tau (p-Tau) and low density lipoprotein receptor-related protein-1(LRP-1) in blood and brain tissues. Methods: In this study, 5-, 10- and 15-month-old APPswe/PS1dE9 double-transgenic mice harboring the genotype of AD confirmed by the gene test were divided into the 5-, 10- and 15-month-old experiment groups, and simultaneously, age-matched C57BL/6J mice were placed into the corresponding control groups, with 8 mice in each group. All mice were subjected to the Morris Water Maze for test of cognitive and behavioral ability. Expression profiles of PTK2B, Aβ1-42, p-Tau/Tau and LRP-1 in the hippocampus or blood of mice were quantified by using the immunohistochemistry staining, Western blot or enzyme-linked immunosorbent assay (ELISA), while the mRNA expression of ptk2b in the hippocampus was quantified by using the real-time quantitative polymerase chain reaction (qRT-PCR). Results: Results of experiment groups demonstrated that as mice aged, the expression levels of PTK2B, ptk2b mRNA, Aβ1-42 and p-Tau/Tau in the hippocampus were increased, and the expression of LRP-1 was decreased gradually. While in the blood, the level of Aβ1-42 was decreased, and the cognitive and behavioral ability was decreased in an age-dependent manner (all P＜ 0.05). However, comparisons among the control groups, only the age-dependent downregulation of LRP-1 were observed in hippocampus(P＜0.05), but other indicators had no significant differences (P＞0.05). Conclusion: In the hippocampus of APP/PS1 double-transgenic mice, the expressions of PTK2B, Aβ1-42 and p-Tau/Tau are upregulated, LRP-1 is downregulated, while cognitive and behavioral ability is decreased, and such changes are presented in a time-dependent manner.

Relationship between Discordance of Low-Density Lipoprotein and Non-High-Density Lipoprotein Cholesterol and Risk Stratification in Acute Myocardial Infarction

Abstract Background: Sizeable proportion of patients have discordant low-density lipoprotein cholesterol (LDL-C) and non-high density lipoprotein cholesterol (NHDL-C). It has been shown that discordance of LDL-C and NHDL-C either underestimates or overestimates coronary risk. Objectıve: We assessed whether this discordance has an impact on GRACE and TIMI risk scores in patients with acute myocardial infarction (AMI). Methods: We retrospectively evaluated the data of 198 consecutive patients with AMI. Fasting serum lipid profiles were recorded, GRACE and TIMI scores were calculated. Patients were divided into 3 groups according to LDL-C and NHDL-C percentiles: Discordant group: LDL-C<NHDL-C (n=38), concordant group: LDL-C=NHDL-C (n=112) and discordant group LDL-C>NHDL-C (n=48). GRACE and TIMI scores, mortality and cardiovascular events (heart failure, non-fatal myocardial infarction and angina) at sixth month were compared between these three groups. Differences between these groups were analyzed with One-way ANOVA or Kruskal-Wallis rank test, and with chi-square for percentages. Also, post hoc LSD or Conover-Iman's non-parametric multiple comparison test were used. A p value <0.05 was accepted as statistically significant. Results: TIMI risk score didn't differ between discordant or concordant groups. Mean GRACE (death) and GRACE (death and MI) scores were higher in group with LDL-C<NHDL-C than with LDL-C=NHDL-C and LDL-C>NHDL-C (p=0.029 and 0.008, respectively). Cardiovascular events and mortality at sixth month were not different among groups (p=0.473 and p=0.176, respectively). Conclusion: GRACE score was higher in discordant group with LDL-C<NHDL-C, but there is no difference regarding TIMI scores between discordant and concordant groups in AMI patients.

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.

Role of the LRP1-pPyk2-MMP9 pathway in hyperoxia-induced lung injury in neonatal rats / 中国当代儿科杂志

OBJECTIVES@#To study the role of the low-density lipoprotein receptor-related protein 1 (LRP1)-proline-rich tyrosine kinase 2 phosphorylation (pPyk2)-matrix metalloproteinases 9 (MMP9) pathway in hyperoxia-induced lung injury in neonatal rats.@*METHODS@#A total of 16 neonatal rats were randomly placed in chambers containing room air (air group) or 95% medical oxygen (hyperoxia group) immediately after birth, with 8 rats in each group. All of the rats were sacrificed on day 8 of life. Hematoxylin and eosin staining was used to observe the pathological changes of lung tissue. ELISA was used to measure the levels of soluble LRP1 (sLRP1) and MMP9 in serum and bronchoalveolar lavage fluid (BALF). Western blot was used to measure the protein expression levels of LRP1, MMP9, Pyk2, and pPyk2 in lung tissue. RT-PCR was used to measure the mRNA expression levels of LRP1 and MMP9 in lung tissue.@*RESULTS@#The hyperoxia group had significantly higher levels of sLRP1 and MMP9 in serum and BALF than the air group (@*CONCLUSIONS@#The activation of the LRP1-pPyk2-MMP9 pathway is enhanced in hyperoxia-induced lung injury in neonatal rats, which may be involved in the pathogenesis of bronchopulmonary dysplasia.

Oxidized low-density lipoprotein stimulates dendritic cells maturation via LOX-1-mediated MAPK/NF-κB pathway

Dendritic cells (DCs) play a crucial role as central orchestrators of immune system response in atherosclerosis initiation and progression. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is involved in the immune maturation of DCs, but the underlying mechanisms remain unclear. We isolated mouse bone marrow progenitors and stimulated them with granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4 to induce immature DCs. We then treated DCs with oxidized low-density lipoprotein (oxLDL) to induce maturation. LOX-1 siRNA was used to investigate the modulation of LOX-1 on the development of DCs and the underlying signal pathways. CD11c-positive DCs were successfully derived from mouse bone marrow progenitors. OxLDL promoted the expressions of DCs maturation markers and pro-inflammatory cytokines. OxLDL also upregulated LOX-1 expression and activated MAPK/NF-κB pathways. LOX-1 siRNA could attenuate the expression of MAPK/NF-κB pathways and inflammatory cytokines. In conclusion, oxLDL induced the maturation of DCs via LOX-1-mediated MAPK/NF-κB pathway, which contributed to the initiation and progression of atherosclerosis.

Effect of Sijunzi Decoction on regulation of Aβ-related proteins across blood-brain barrier / 中国中药杂志

According to traditional Chinese medicine, "spleen transport" is closely related to the metabolism of substance and energy. Studies have shown that Alzheimer's disease(AD) is a disease related to glucose and lipid metabolism and energy metabolism. The traditional Chinese medicine Jiangpi Recipe can improve the learning ability and memory of AD animal model. Sijunzi Decoction originated from Taiping Huimin Hefang Prescription is the basic prescription for strengthening and nourishing the spleen, with the effects of nourishing Qi and strengthening the spleen. In this experiment, human brain microvascular endothelial cells(HBMEC) and Sijunzi Decoction water extract(0.25, 0.5, 1 mg·L~(-1)) were pre-incubated for 2 h, and then Aβ_(25-35) oligomers(final concentration 40 μmol·L~(-1)) was added for co-culture for 22 hours. The effect of Sijunzi Decoction on the activity of Aβ_(25-35) oligomer injured cells and the expression of related proteins were investigated. Q-TOF-LC-MS was used first for principal component analysis of Sijunzi Decoction water extract. Then MTT assay was used to investigate the effect of Sijunzi Decoction water extract on the proliferation of HBMEC cells. Real-time fluorescence quantitative PCR(RT-qPCR) was employed to detect the mRNA expression of GLUT1, RAGE, and LRP1. The expression of Aβ-related proteins across blood-brain barrier(RAGE, LRP1) was detected by Western blot. The results showed that 40 μmol·L~(-1) Aβ_(25-35) oligomers could induce endothelial cell damage, reduce cell survival, increase expression of RAGE mRNA and RAGE protein, and reduce expression of GLUT1 mRNA, LRP1 mRNA, and LRP1 protein. Sijunzi Decoction water extract could reduce the Aβ_(25-35) oligomer-induced cytotoxicity of HBMEC, decrease the expression of RAGE mRNA and RAGE protein, and increase the expression of GLUT1 mRNA, LRP1 mRNA and LRP1 protein. The results indicated that Sijunzi Decoction could reduce the injury of HBMEC cells induced by Aβ_(25-35) oligomer, and regulate the transport-related proteins GLUT1, RAGE and LRP1, which might be the mechanism of regulating Aβ_(25-35) transport across the blood-brain barrier.

One family with osteosclerosis caused by D111Y mutation in the low-density lipoprotein receptor-related protein 5 gene / 中华内分泌代谢杂志

Objective@#To investigate the clinical features and pathogenic genes of a family with osteosclerosis.@*Methods@#Six patients and six family members from a family in Jiangsu were tested for biochemical parameters, bone metabolic markers, bone mineral density, thoracolumbar anterior lateral slices, skull positive lateral radiographs, and pelvic plain films. Meanwhile, Sanger sequencing was performed to detect gene mutations of the proband and five other family members with high bone mass. The conformation of the mutational low-density lipoprotein receptor-related protein 5 (LRP5) protein was predicted by SWISS-MODEL.@*Results@#Four adult patients (one male and three females) were tall, with mandibular enlargement and kyphosis in the center of the lower jaw, and none of the four had fractures. Their X ray examination revealed that the skull and long bone cortex was thickened, while the sella and mandible was enlarged. In addition, the absolute values of bone mineral density at each site of all patients were significantly higher as compared with the standard age- and sex-matched adults or adolescent mean reference values, with Z scores of L2-4, femoral neck and total hip being (6.31±4.03) SD, (6.56±2.36) SD, and (7.19±2.03) SD, respectively. The results of genetic sequencing revealed that all six patients carried a heterozygous mutation (c.331G>T; D111Y) in exon 2 of LRP5 gene, while other family members showed wild type (c.331G>G; D111D). Functional prediction indicated that this mutation was located at the amino acid terminal of exon 2 of LRP5 gene, which encodes the first β-helix-generating region of LRP5 protein.@*Conclusion@#The D111Y mutation in LRP5 gene leads to a clinical phenotype characterized by benign increased bone mineral density without increasing the risk of fracture. This mutation may further affect the downstream Wnt signaling pathway by altering the spatial structure of LRP5 protein, thereby promoting maturation and differentiation of osteoblasts and resulting in osteosclerosis.

Effect of Evodia on Lipid Metabolism in Hyperlipidemia Mice / 中国实验方剂学杂志

Objective::To study the effect of evodia on lipid metabolism and low-density lipoprotein-receptor(LDL-R) mRNA expression in hyperlipidemia mice. Method::Kunming mice (n=80) were randomly divided into normal control group (n=20) and model group (n=60). Serum lipids of the model group were measured after 3 weeks.After successful modeling, the mice can be randomly divided into 5 groups (with 10 in each group): model group (equivalent normal saline), positive control group (simvastatin, 5 mg·kg-1·d-1), drug group (evodia of 5.25, 10.5, 21 mg·kg- 1·d- 1). The mice were given drugs for 3 weeks.Htoxylin-eosin(HE) staining was used to observe the liver cell structure and the change of aortic arch atherosclerosis in the mice.The enzyme linked immunosorbent assay kit was used to test the contents of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and total serum adiponectin (ADPN) in serum of the mice.The expression of LDL-R mRNA in liver of each group was detected by reverse transcription-polymerase chain reaction (RT-PCR). Result::Liver HE staining showed hepatocyte swelling with steatosis in the model group, and alleviated liver steatosis in high-dose, medium-dose evodia and simvastatin groups.HE staining showed damages on the aortict arch wall in the model group, with obvious intima thickening and inflammatory cell infiltration.The intima was thickened obviously in the low-dose group, and the structure of aortic vessel wall was clear in the high-dose group.Compared with the normal group, TC, TG and HDL-C levels in serum of the model group were increased, while HDL-C level was decreased (P<0.01). Serum TC and TG levels of mice in the medium and high-dose groups decreased, whereas LDL-C and HDLl-C levels increased in low, medium and high-dose groups (P<0.05, P<0.01). Compared with the normal group, the adiponectin level in the model group was decreased, while the serum adiponectin levels in medium and high-dose groups were significantly increased (P<0.01). The LDL-R mRNA expression in the liver of mice in the model group was significantly reduced compared with the normal group (P<0.01). The LDL-R mRNA expression in medium and high-dose evodia groups was significantly increased compared with the model group (P<0.01). Conclusion::Evodia can improve the tendency of hepatic lesions and aortic atherosclerosis in hyperlipidemia mice, which may be related to the regulation of adiponectin level, the reduction of lipid content in mice and the up-regulation of LDL-R mRNA expression in mice liver.

Effect of Zhenxin Shengshui Yizhi Fang on Damage of Human Brain Microvascular Endothelial Cells Induced by Aβ25-35 / 中国实验方剂学杂志

Objective::To observe the neuroprotective effect and potential mechanism of Zhenxin Shengshui Yizhi Fang(XSF) aqueous extract on human brain microvascular endothelial cells (HBMEC) injury induced by amyloid-β protein(Aβ)25-35. Method::HBMEC cells damage induced by Aβ25-35 was used as Alzheimer' s disease(AD) cell model. The study included control group, Aβ25-35 group, and low, medium and high-dose XSF aqueous extract groups (125, 250, 500 mg·L-1). After treatment, the cytotoxicity of different concentrations of drugs and Aβ25-35 was determined by methyl thiazolyl tetrazolium(MTT) colorimetry. Apoptosis was observed by Hoechst-33258 staining. The activity of Caspase-3 was detected by colorimetry. Western blot was used to detect the expression levels of the receptor of advanced glycation end products (RAGE) and low-density lipoprotein receptor-related protein (LRP1). Result::Compared with the control group, the cell viability of Aβ25-35 group was significantly decreased (P<0.01). Hoechst-33258 staining showed bright blue fluorescence, chromatin condensation, dense staining or fragmentation dense staining, whitening in color, and significant increase of the percentage of apoptotic cells (P<0.01). Caspase-3 activity increased significantly (P<0.01). Western blot showed that RAGE protein expression increased significantly (P<0.01), while low-density lipoprotein receptor-related protein(LRP1), glucose transporter 1(GLUT1) and GLUT3 protein expressions decreased significantly (P<0.01). Compared with the Aβ25-35 group, the cell viability of XSF aqueous extract groups was significantly increased in a dose-dependent manner. The XSF aqueous extract had a more significant protective effect of than the other groups (P<0.05). The XSF aqueous extract group (500 mg·L-1) significantly inhibited the number of apoptotic cells (P<0.01), but significantly reduced the Caspase-3 activity (P<0.01). RAGE protein expression was not significantly decreased in XSF aqueous extract group (125 mg·L-1), but significantly decreased in XSF aqueous extract group (250, 500 mg·L-1, P<0.01), while LRP1, GLUT1 and GLUT3 protein expression significantly increased (P<0.05, P<0.01) in a dose-dependent manner. Conclusion::XSF aqueous extract can attenuate the cytotoxicity of HBMEC induced by Aβ25-35 oligomer, inhibit apoptosis, decrease caspase-3 activity and RAGE protein expression, increase LRP1, GLUT1 and GLUT3 protein expressions, and reduce the abnormal accumulation and deposition of Aβ in the brain, which may be its mechanisms for prevention and treatment of AD.

Effect of aerobic exercise on blood lipid of hyperlipidemia rats and its mechanism / 第二军医大学学报

Objective To explore the effect of aerobic exercise on blood lipid of hyperlipidemia rats and its mechanism. Methods A total of 120 healthy male SD rats aged 8 weeks were randomized into normal control group, high-fat diet (HF) group, SBC-115076 group and aerobic exercise group, with 30 rats in each group. The rats in the normal control group were fed with standard diet, and the rats in the other 3 groups were fed with HF to establish hyperlipidemia rat model. The rats in the SBC-115076 group were injected with proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitor SBC-115076 (8 mg/kg) once a week for 8 weeks, and the rats in the aerobic exercise group underwent swimming without load 6 days a week for 8 weeks. After 8 weeks, the rats were sacrificed and the blood samples were collected to determine the levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL) and high-density lipoprotein (HDL). Pathological changes of thoracic aorta were observed using H-E staining. The mRNA and protein expression levels of PCSK9, low-density lipoprotein receptor (LDLR) and sterol regulatory element binding protein (SREBP) in the hepatic tissues were detected by quantitative real-time PCR, Western blotting and immunofluorescence. Results Compared with the normal control group, the levels of serum TG, TC and LDL of the rats were significantly higher in the HF group, and the level of HDL was significantly lower (P<0.01). Compared with the HF group, the levels of serum TG, TC and LDL of the rats were significantly lower in the SBC-115076 and aerobic exercise groups, and the level of HDL was significantly higher (P<0.01). In the HF rats, the aortic tunica intima was thickened and endothelial cells were damaged and exfoliated. Compared with the HF group, the aortic intima thickening was reduced and endothelial damage was less in the aerobic exercise group. Compared with the normal control group, the mRNA and protein expression levels of PCSK9, SREBP1 and SREBP2 were significantly higher in the HF group, and the mRNA and protein expression levels of LDLR were significantly lower (P<0.01). Compared with the HF group, the mRNA and protein expression levels of PCSK9, SREBP1 and SREBP2 were significantly lower, and the mRNA and protein expression levels of LDLR were significantly higher (P<0.01). Conclusion Aerobic exercise can down-regulate the expression of TG, TC and LDL, up-regulate the expression of HDL, and alleviate the intimal thickening of aorta. The mechanism may be related to down-regulating the expression of PCSK9 and SREBPs, thus eliminating the inhibition of LDLR.

Changes and clinical significance of serum sLOX-1 and omentin-1 in patients with H-type hypertension complicated with acute ischemic stroke / 实用医学杂志

Objective To investigate the changes of soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) and omentin-1 in patients with H-type hypertension complicated with acute ischemic stroke, and to analyze the correlation of sLOX-1 and omentin-1 levels with the severity and prognosis of the disease.Methods Totally 136 patients with H-type hypertension complicated with acute ischemic stroke from February2017 to May 2018 were selected as observation group, and 136 non-acute ischemic stroke patients with H-type hypertension in the same period as the control group. The patients of observation group were divided into mild, moderate and severe sub-groups according to NIHSS score, and they were also divided into good prognosis group and poor prognosis group based on modified RANKIN scale (mRS) score. The serum sLOX-1 and omentin-1 levels were detected, and the correlation of sLOX-1 and omentin-1 levels with severity and prognosis of disease was analyzed. Results The serum sLOX-1 level of the observation group was higher, but the serum omentin-1 level lower than that of control group (P < 0.05). With the severity of the disease, the serum sLOX-1 level increased, but the serum omentin-1 level decreased (P < 0.05). The serum sLOX-1 level of good prognosis group was significantly lower, whereas the serum omentin-1 level significantly higher than that of poor prognosis group (P < 0.05). sLOX-1 was positively correlated with NIHSS score and mRS score, while omentin-1 was negatively correlated with NIHSS score and mRS score (P < 0.05). Conclusions The levels of serum sLOX-1 and omentin-1 are closely related to the severity and prognosis of patients with H-type hypertension complicated with acute ischemic stroke, which could be used as markers for evaluating the severity and prognosis of the patients.

The hypolipidemic and anti-atherosclerotic effects of acacetin and its mechanism of action in mice / 药学学报

The purpose of this research is to investigate the effects of acacetin on serum lipid metabolism and atherosclerosis in mice and explore its molecular mechanism. HepG2 cells were treated with different concentrations of acacetin. The expression of LDL receptor (LDLR) and sterol-regulatory element binding protein-2 (SREBP-2) were detected by RT-qPCR and/or Western blot. C57BL/6J mice were given acacetin (50 mg·kg-1) for 5 weeks by gavage. Serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and triglyceride (TG) were analyzed by an automatic biochemical analyzer. The expression of LDLR or SREBP-2 was detected by Western blot. After 12 weeks of intragastric administration of acacetin (30 mg·kg-1) in apolipoprotein E knockout (ApoE KO) mice, the serum lipid levels were determined by an automatic biochemical analyzer. The lipid deposition in aortic plaque (en face) and aortic root plaque were stained with oil red O. The expression of LDLR and SREBP-2 were detected by RT-qPCR and/or Western blot. The intestinal content microflora was analyzed by 16S rDNA sequencing (All animal studies were approved by the Animal Experimentation Ethics Committee of Institute of Medicinal Biotechnology, CAMS & PUMC). In vitro results indicated that acacetin significantly up-regulated LDLR mRNA and protein levels, and stimulated LDLR transcription factor SREBP-2 protein expression. As indicated from in vivo studies, compared with control group, acacetin significantly decreased the serum levels of TC and LDL-C in C57BL/6J mice by 34% and 57% (P<0.01), respectively. Furthermore, mechanic study showed that acacetin significantly increased the protein expression of hepatic LDLR and SREBP-2. Although the results of serum lipid profiles, hepatic LDLR/SREBP-2 expression and area of atherosclerotic lesions in aorta and aortic root in ApoE KO mice showed differences between acacetin and high-fat diet group, the differences did not reach statistical significance. Nevertheless, acacetin exhibited a profound influence on the composition of the intestinal microbiota as indicated by 16s rDNA sequencing analysis. In conclusion, these results demonstrated that acacetin can decrease the serum lipid levels in C57BL/6J mice through up-regulation of hepatic LDLR and SREBP-2, and alter gut microflora in high-fat diet fed Apo KO mice. This study suggests the possibility that acacetin has a potential role in inhibiting the progression of atherosclerosis.

Clinical research of PCSK9 inhibitors in reducing low density lipoprotein cholesterol / 上海交通大学学报（医学版）

Elevated low density lipoprotein cholesterol (LDL-C) is a major risk factor for cardiovascular disease. Studies show that proprotein convertase subtilisin/kexin type 9 (PCSK9) is a circulation enzyme and serves a pivotal function in the degradation of low density lipoprotein receptor, which contributes to the decrease in hepatic cholesterol uptake and increase in circulating LDL-C. PCSK9 inhibitor can significantly elevate the surface of low density lipoprotein receptor of liver cells and bond more LDL-C to decrease the level of LDL-C. Thus PCSK9 has emerged as a popular target for the development of new cholesterol lowering drugs and therapeutic intervention of cardiovascular disease. In this article, the history, mechanism of action, metabolic effects of PCSK9 and the clinical outcomes of PCSK9 inhibitors will be briefly reviewed.

Effects of high-fat diet on bone mineral density and microarchitecture in LDLR-/-mice / 中华内分泌代谢杂志

Objective To investigate the effect of high-fat diet on bone calcium content, bone mineral density,and microarchitecture in low density lipoprotein receptor knockout(LDLR-/-) mice. Methods Thirteen LDLR-/- mice aged 11 weeks and 13 sex-age-matched C57BL/6J mice(WT)were divided into WT group with normal diet,WT group with high-fat diet,LDLR-/- group with normal diet,and LDLR-/- group with high-fat diet. All mice were sacrificed at 40 weeks of age and left tibias were harvested to analyze the bone mineral density and bone microarchitecture parameters. The right tibias were sampled for measurement of osteoporogeterin(OPG)and receptor activator of nuclear factor kappa-B ligand(RANKL)mRNA expressions using real-time PCR. Results LDLR-/-and high-fat diet significantly changed bone calcium content, bone microarchitecture parameters, and OPG/RANKL mRNA ratio(P<0.05). Compared with WT mice, LDLR-/-mice with high-fat diet showed a lower bone calcium content,along with correspondingly decreased parameters of bone microarchitecture(P<0.05). Compared with WT mice with normal diet, bone calcium content, tissue bone mineral density, and OPG/RANKL mRNA ratio were decreased in LDLR-/-mice with ordinary diet(P<0.01),while no change of other bone microarchitecture parameters was found. Conclusions High-fat diet results in decreased bone mass and damaged bone microarchitecture in LDLR-/- mice,which may be attributed to the down-regulation of OPG/RANKL mRNA ratio.