Berberine Inhibits Atherosclerosis by Regulating Lipophagy Via Targeting Wnt5a/NPC1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect and molecular mechanism of berberine （BBR） on lipophagy in the prevention and treatment of atherosclerotic （AS） lesions in mice. MethodFifty apolipoprotein E-knockout （ApoE-/-） mice were randomly divided into an AS model group， an atorvastatin group （5 mg·kg-1）， and low-， medium-， and high-dose BBR groups （2.5， 5， 10 mg·kg-1）. Ten C57BL/6J mice were assigned to the control group. After 12 weeks， hematoxylin-eosin （HE） and oil red O staining were performed to assess the histopathological changes of AS plaques in the aorta. Biochemical analysis was used to measure serum lipid levels， and enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of inflammatory cytokines interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）， oxidative stress marker reactive oxygen species （ROS）， and serum lipophagy marker Beclin1 and microtubule-associated protein 1 light chain 3 Ⅱ （LC3Ⅱ）. The xanthine oxidase method was used to measure serum superoxide dismutase （SOD） activity. Immunohistochemistry （IHC） was used to detect the distribution of wingless-type MMTV integration site family member 5a （Wnt5a） and Nieman Pick type C1 （NPC1） in the aorta， and Western blot was used to determine the protein expression of Wnt5a and NPC1 in the aorta. ResultCompared with the control group， the AS model group showed significant AS plaque formation， significantly elevated levels of serum total cholesterol （TC）， triglycerides （TG）， low-density lipoprotein cholesterol （LDL-C）， IL-6， TNF-α， and ROS， aortic Wnt5a distribution and protein expression （P<0.01）， and significantly reduced levels of serum high-density lipoprotein cholesterol （HDL-C）， SOD， Beclin1， LC3Ⅱ， and aortic NPC1 distribution and protein expression （P<0.01）. Compared with the AS model group， the atorvastatin group， and high- and medium-dose BBR groups showed a significant reduction in AS plaque area （P<0.05， P<0.01）， significantly decreased levels of serum TC， TG， LDL-C， IL-6， TNF-α， ROS， and aortic Wnt5a distribution and protein expression （P<0.05， P<0.01）， and significantly increased levels of serum HDL-C， SOD， Beclin1， LC3Ⅱ， and aortic NPC1 distribution and protein expression （P<0.05， P<0.01）. There was no statistically significant difference in the above indicators between the atorvastatin group and the medium-dose BBR group. ConclusionBBR can competitively bind to Wnt5a to activate NPC1 expression， upregulate lipophagy levels， reduce blood lipids， and inhibit the release of inflammatory mediators and oxidative stress damage， thereby exerting a preventive and therapeutic effect on AS.

Cathepsin D overexpression in the nervous system rescues lethality and Aβ42 accumulation of cathepsin D systemic knockout in vivo

The lysosome is responsible for protein and organelle degradation and homeostasis and the cathepsins play a key role in maintaining protein quality control. Cathepsin D (CTSD), is one such lysosomal protease, which when deficient in humans lead to neurolipofuscinosis (NCL) and is important in removing toxic protein aggregates. Prior studies demonstrated that CTSD germ-line knockout-CtsdKO (CDKO) resulted in accumulation of protein aggregates, decreased proteasomal activities, and postnatal lethality on Day 26 ± 1. Overexpression of wildtype CTSD, but not cathepsin B, L or mutant CTSD, decreased α-synuclein toxicity in worms and mammalian cells. In this study we generated a mouse line expressing human CTSD with a floxed STOP cassette between the ubiquitous CAG promoter and the cDNA. After crossing with Nestin-cre, the STOP cassette is deleted in NESTIN + cells to allow CTSD overexpression-CTSDtg (CDtg). The CDtg mice exhibited normal behavior and similar sensitivity to sub-chronic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced neurodegeneration. By breeding CDtg mice with CDKO mice, we found that over-expression of CTSD extended the lifespan of the CDKO mice, partially rescued proteasomal deficits and the accumulation of Aβ42 in the CDKO. This new transgenic mouse provides supports for the key role of CTSD in protecting against proteotoxicity and offers a new model to study the role of CTSD enhancement in vivo.

Simvastatin alleviates liver function injury in ApoE knockout mice by inhibiting calpain activity / 中国药理学通报

Aim To study the protective effect of simvastatin(Sim)on liver function injury in apolipoprotein E gene knockout(ApoE KO)mice fed with high-fat diet and the underlying mechanism.Methods Twenty-four 8-week-old male ApoE KO mice were randomly divided into ApoE KO group,ApoE KO+Sim group and ApoE KO+PD150606 group.The contents of total cholesterol(TC)and triglyceride(TG)in serum and liver,and the activities of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in serum were measured.The contents of malondialdehyde(MDA)and reactive oxygen species(ROS)and the activity of superoxide dismutase(SOD)in liver were determined.The contents of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)and the activity of calpain in liver were examined.Results Compared with C57 group,ApoE KO group showed significant increase in the contents of TC and TG in both serum and liver.In addition,the activities of AST and ALT in serum and the contents of MDA and ROS in liver significantly increased,while SOD activity in liver decreased in ApoE KO group.The contents of TNF-α and IL-6 and the activity of calpain in liver significantly increased.Compared with ApoE KO group,Sim group had no significant effects on TC and TG,while reduced the activities of AST and ALT,decreased the contents of MDA and ROS,increased the activity of SOD and decreased the contents of TNF-α and IL-6 as well as the activity of calpain in liver.PD,the calpain inhibitor,had the similar effects with Sim regarding the above mentioned parameters.Conclusions Sim improved the liver function injury of ApoE KO mice,which might be related to the inhibition of calpain activity,subsequently increasing the antioxidant capacity and reducing the inflammatory response.

Quercetin attenuates atherosclerosis in ApoE KO mice by regulating macrophage cholesterol homeostasis / 中国药理学通报

Aim To investigate the protective effect of quercetin on atherosclerosis induced by high-fat diet in ApoE knockout ( ApoE KO) mice and its regulatory mechanism on cholesterol homeostasis of macrophages.Methods Forty-five adult female ApoE KO mice were randomly divicied into three groups : nonnal diet ( ND ) group, high fat diet ( HFD) group and high fat diet + quercetin ( HFD + Qu) group and fed for 16 weeks.The level of serum lipid, the formation of atherosclerotic plaque and the expression of genes related to cholesterol homeostasis were detected.Macrophage cholesterol content and the expression level of cholesterol homeo- stasis-related proteins were detected.Results Quer cetin significantly reduced the atherosclerotic lesions and serum lipid levels in ApoE KO mice.Quercetin significantly suppressed macrophage foaming by upreg- ulating CYP27A1 expression，inhibiting CD36-mediated cholesterol uptake and and promoting LXHcx-ABCAl/ G1 pathway-dependent cholesterol efflux.Conclusions Quercetin plays a protective role in atherosclerosis through its regulatory effect on CYF27A1/ LXHa signaling pathway-mediated macrophage cholesterol homeostasis.

CRISPR/Cas9-mediated genome editing reveals 12 testis-enriched genes dispensable for male fertility in mice / 亚洲男科学杂志(英文版)

Gene expression analyses suggest that more than 1000-2000 genes are expressed predominantly in mouse and human testes. Although functional analyses of hundreds of these genes have been performed, there are still many testis-enriched genes whose functions remain unexplored. Analyzing gene function using knockout (KO) mice is a powerful tool to discern if the gene of interest is essential for sperm formation, function, and male fertility in vivo. In this study, we generated KO mice for 12 testis-enriched genes, 1700057G04Rik, 4921539E11Rik, 4930558C23Rik, Cby2, Ldhal6b, Rasef, Slc25a2, Slc25a41, Smim8, Smim9, Tmem210, and Tomm20l, using the clustered regularly interspaced short palindromic repeats /CRISPR-associated protein 9 (CRISPR/Cas9) system. We designed two gRNAs for each gene to excise almost all the protein-coding regions to ensure that the deletions in these genes result in a null mutation. Mating tests of KO mice reveal that these 12 genes are not essential for male fertility, at least when individually ablated, and not together with other potentially compensatory paralogous genes. Our results could prevent other laboratories from expending duplicative effort generating KO mice, for which no apparent phenotype exists.

Impact of angiotensin-converting enzyme inhibition on hemodynamic and autonomic profile of elastase-2 knockout mice

Elastase-2 (ELA-2) is an angiotensin II-generating enzyme that participates in the cardiovascular system. ELA-2 is involved in hemodynamic and autonomic control and is upregulated in myocardial infarction and hypertension. The inhibition of angiotensin-converting enzyme (ACE) increased ELA-2 expression in the carotid arteries and heart of spontaneously hypertensive rats. In this study, we sought to investigate the role of ACE inhibition in hemodynamic and autonomic balance in elastase-2 knockout (ELA-2 KO) mice. Male ELA-2 KO and C57BL/6 mice were treated with the ACE inhibitor enalapril or saline for 10 days. After treatment, mice underwent surgery for cannulation of the femoral artery and arterial pressure recordings were made five days later in awake animals. The variability of systolic blood pressure (SBP) and pulse interval (PI) was evaluated in the time and frequency domain. Spontaneous baroreflex was assessed by the sequencing method. ACE inhibition caused a significant decrease in mean arterial pressure (117±2.2 vs 100±2.8 mmHg) and an increase in heart rate (570±32 vs 655±15 bpm) in ELA-2 KO mice. Despite a tendency towards reduction in the overall heart rate variability (standard deviation of successive values: 7.6±1.1 vs 4.7±0.6 ms, P=0.08), no changes were found in the root of the mean sum of squares or in the power of the high-frequency band. ACE inhibition did not change the spontaneous baroreflex indices (gain and baroreflex effectiveness index) in ELA-2 KO mice. Altogether, this data suggested that ACE played a role in the maintenance of hemodynamic function in ELA-2 KO mice.

Role of Ca / 中国药理学通报

Aim To investigate the role of Ca

Sevoflurane preconditioning protects myocardial ischemia/reperfusion injury by increasing plasma adiponectin levels / 中国药理学通报

Aim To investigate whether adiponectin is involved in the protective effect of sevoflurane preconditioning on myocardial ischemia/reperfusion (MI/R) injury. Methods C57 mice were randomly divided into four groups; the Sham Control (sham group), the MI/R-Control (model group), the MI/R-Sevopre (pretreatment group) and the APN-KO-MI/R-Sevopre (knockout pretreatment group). The plasma adiponectin level was detected 24 h after MI/R modeling. Meanwhile, the left ventricular end-systolic diameter, end-diastolic diameter and ejection fraction were measured at 24 h. In addition, TUNEL staining was used to detect the left ventricular end-systolic diameter, end-diastolic diameter and ejection fraction and the morphology and apoptosis of myocardial cells in each group were observed. What' s more, the infarct area was observed by Evans blue TTC staining. Results Compared with sham group, model group had impaired heart function, decreased ejection fraction and increased end-diastolic and end-systolic diameters. Meanwhile, the myocardial infarction area and apoptotic cells significantly increased in model group, and the plasma adiponectin level decreased. However, compared with model group, mice in sevoflurane pretreatment group had improved heart function, decreased myocardial infarction area, decreased apoptotic cells and increased plasma adiponectin levels. And then, compared with pretreatment group, sevoflurane pretreatment in knockout pretreatment group reduced Ml/R injury. Conclusion The sevoflurane pretreatment protects MI/R injury by increasing plasma adiponectin levels.

The effects of casein kinase 2 interacting protein-1 on the growth and development of craniomaxillofacial soft and hard tissues in mice / 口腔疾病防治

Objective@# To investigate the effect of casein kinase 2 interacting protein-1 (CKIP-1) on craniofacial soft tissues and hard tissues, to provide the basis for the study and treatment of craniomaxillofacial related diseases.@*Methods@#6-month- old male CKIP-1 knockout (KO) mice were selected as the experimental group, and wild-type (WT) mice were selected as the control group. The craniomaxillofacial hard tissues (parietal bone, nasal bone, incisors and molars) were analyzed through micro- CT, and the morphological changes of maxillofacial soft tissues (nasal cartilage, lip mucosa and tongue) were analyzed through HE staining and toluidine blue staining.@* Results@#CKIP-1 negatively regulated bone mass of cancellous bone of cranial and maxillofacial bones and dentin mineralization. Compared with the WT mice, the thickness of the parietal baffle layer increased by 93% in KO mice, while cortical bone showed no significant difference between the two groups. The nasal cancellous bone thickness increased by 160% in KO-mice, while cortical bone showed no significant difference between the two groups; the enamel thickness was normal, but the pulp cavity became smaller and the dentin thickness increased by 48%. Compared with the WT mice, the HE staining and toluidine blue staining analyses of the soft tissues revealed that the thickness of the alar cartilage plate of KO mice increased by 57%, and local ossification was found within the cartilage plate. The thickness of the keratinized layer of the labial mucosa increased by 170% in KO mice and the muscle fiber diameter of the lingual muscle increased by 45%. @*Conclusion@#CKIP-1 genes have different effects on the growth and development of various soft and hard tissues in the maxillofacial region of mice.

Construction of β2m knockout mice / 生物工程学报

The β2m (Beta-2-microglobin) gene encodes a non-glycosylated protein that functions as an important component of major histocompatibility complexⅠ(MHCⅠ) for antigen presentation. To evade immune mediated clearance, human tumors and pathogens have adopted different strategies, including loss of MHCⅠexpression. Appropriate animal models are essential for understanding the mechanisms underpinning the clinical treatment of tumor and other human diseases. We constructed β2m knockout mice using CRISPR/Cas9 gene editing tool through embryo microinjection. Subsequently, genotyping and phenotyping of knockout mice were performed by PCR, qPCR, and flow cytometry. Mice genotyping showed that the coding region of the target gene was absent in the knockout mice. Real time PCR showed that mRNA level of β2m was significantly downregulated. Flow cytometry showed that the proportions of CD8+ killer T cells was significantly reduced in a variety of tissues and organs of the immune system. Taken together, we have successfully constructed a strain of β2m knockout mice, which will facilitate subsequent in vivo study on the function and mechanism of the β2m gene.

Pathogenic role of NDUFA13 inactivation in spontaneous hepatitis in mice and the mechanism / 南方医科大学学报

OBJECTIVE@#To investigate the role of NDUFA13 inactivation in the pathogenesis of spontaneous hepatitis in mice and explore the possible mechanisms.@*METHODS@#Hepatocyte-specific NDUFA13 knockout (NDUFA13@*RESULTS@#Liver-specific NDUFA13 heterozygous knockout mice were successfully constructed as verified by PCR results. HE staining revealed severe liver damage in both 4- week-old and 2-year-old NDUFA13@*CONCLUSIONS@#Hepatocytes-specific NDUFA13 ablation can trigger spontaneous hepatitis in mice possibly mediated by the activation of ROS/NF-κB/NLRP3 signaling.

Naringenin promotes cell autophagy to improve high-fat-diet-induced atherosclerosis in ApoE-/- mice

Naringenin (NAR) is a major flavanone in citrus fruits that has multiple pharmacological attributes such as anticancer and antiatherogenic. This study aims to investigate the mechanism of NAR in high-fat-diet (HFD)-induced atherosclerosis (AS) in apolipoprotein E-knockout (ApoE-/-) mice. A HFD-induced AS ApoE-/- mouse model was established. The mice were treated with HFD, different doses of NAR and simvastatin (Simv). After drug treatment, the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD), and alanine aminotransferase (ALT) were determined. The expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected using qRT-PCR and enzyme-linked immunosorbent assay. The plaque area of the aorta of AS mice was determined using oil red O staining. Western blot analysis was applied to measure the levels of autophagy-related proteins [protein 1 light chain 3B (LC3B), beclin 1, and p62]. The TC, TG, LDL-C, TNF-α, ALT, and MDA levels were significantly increased while the HDL-C, SOD, and GSH-Px levels were decreased in the HFD-induced AS ApoE-/- mice. NAR treatment reversed the expression of the above indicators in mice. After they were treated with different doses of NAR, the LC3B and beclin 1 levels were improved while the p62 protein level was decreased. This study suggested that NAR could promote cell autophagy to improve HFD-induced AS in ApoE-/- mice.

Effect of Hic-5 gene knockout on NF-κB/p65 expression and CCl4-induced liver fibrosis degree in mice / 临床肝胆病杂志

@#ObjectiveTo investigate the effect of Hic-5 gene knockout on NF-κB/p65 expression and liver fibrosis. MethodsTen wild-type male C57BL/6 mice were randomly divided into wild-type control group (WT-Control group with 5 mice) and wild-type experimental group (WT-CCl4 group with 5 mice), and ten male C57BL/6 mice with Hic-5 gene knockout were randomly divided into Hic-5 knockout control group (Hic-5 KO-Control group with 5 mice) and Hic-5 knockout experimental group (Hic-5 KO-CCl4 group with 5 mice). The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. Picrosirius red staining was used to observe collagen deposition in liver tissue. Immunohistochemistry was used to measure the expression of alpha-smooth muscle actin (α-SMA) and p65 protein, and real-time quantitative PCR was used to measure the mRNA expression of α-SMA, collagen 1, and p65 in liver tissue. The primary hepatic stellate cells of mice were isolated and stimulated with different concentrations of TGF-β1, and then real-time quantitative PCR was used to measure the mRNA expression of α-SMA, collagen 1, and p65 in primary hepatic stellate cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsPicrosirius red staining showed that compared with the WT-CCl4 group, the Hic-5 KO-CCl4 group had a significant reduction in collagen fibers in liver tissue (P＜0.001). Measurement of serum ALT and AST showed that there were significant differences in ALT and AST between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=22.85 and 25.15, both P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower serum levels of ALT and AST than the WT-CCl4 group (both P＜0.05). Immunohistochemistry showed that there were significant differences in the expression levels of α-SMA and p65 protein in liver tissue between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=207.10 and 98.16, both P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower expression of α-SMA and p65 protein in liver tissue than the WT-CCl4 group (both P＜0.01). The results of real-time quantitative PCR showed that there were significant differences in the relative mRNA expression of α-SMA, collagen 1, and p65 in liver tissue between the WT-Control group, the WT-CCl4 group, the Hic-5 KO-Control group, and the Hic-5 KO-CCl4 group (F=41.62, 13.93, and 98.16, all P＜0.001), and the Hic-5 KO-CCl4 group had significantly lower relative mRNA expression of α-SMA, collagen 1, and p65 in liver tissue than the WT-CCl4 group (all P＜0.05). After the primary hepatic stellate cells were stimulated by TGF-β1 at concentrations of 0, 5, and 10 ng/ml, there were significant differences in the relative mRNA expression of α-SMA, collagen 1, and p65 between the WT 0 ng/ml group, the WT 5 ng/ml group, the WT 10 ng/ml group, the KO 0 ng/ml group, the KO 5 ng/ml group, and the KO 10 ng/ml group (F=53.9, 75.82, and 52.41, all P＜0.001), and the Hic-5 KO group had significantly lower relative mRNA expression of α-SMA, collagen 1, and p65 than the WT group (all P＜0.01). ConclusionHic-5 knockout inhibits NF-κB/p65 expression and hepatic stellate cell activation and alleviates CCl4-induced liver fibrosis.

Generation and identification of carbohydrate response element-binding protein conditional gene knockout mouse model / 第二军医大学学报

Objective To establish a conditional knockout mouse model of transcription factor carbohydrate response element-binding protein (ChREBP), so as to provide a technical approach for exploring the biological function of ChREBP in vivo. Methods and results Using Cre/loxP gene targeting strategy, we constructed the gene targeting vector and franked the 8th exon of ChREBP gene with loxP site in embryonic stem cells (ES cells) via homologous recombination. We microinjected the recombined ES clones into blastocysts and implanted into the uterus of pseudopregnant mice to obtain the chimeric mice and ChREBPflox/+ mice. By crossbreeding ChREBPflox/+ mice with Alb-Cre mice, we generated liver-specific ChREBP knockout mice. Conclusion The generation of ChREBP conditional knockout mouse model provides an important means to reveal the physiological function and pathological significance of ChREBP in different tissues.

Mrp1 is up-regulated in lung tissues of mildly stable COPD mice induced by smoke, Nrf2 pathway may be involved / 中国药理学通报

Aim: To investigate the effect of Nrf2 pathway on the expression of MRP1 in mildly stable COPD mice. Methods: The mild COPD mouse model was established by passive cigarette smoking. The pathological changes of lung tissues were examined by HE staining. Immunohistochemistry and Western blot were used to detect the protein expression of MRP1, Nrf2 and HO-1. Results: Compared with normal group, each lung function index of the mild-moderate COPD model group was significantly lower, but compared with wide type(WT) model group, the reduction was more significant in Nrf2-/- model group. HE results showed diffuse inflammatory reaction and alveolar bronchial structure damage in alveolar of WT and Nrf2-/- model mice, and it was more pronounced in Nrf2-/- mice. Immunohistochemistry and Western blot results showed that the expression of MRP1 in lung tissue of Nrf2-/- normal mice was significantly reduced compared with the normal WT group. After passive cigarette smoking, The expression of MRP1, Nrf2 and HO-1 in WT model group increased significantly, but compared with Nrf2-/- normal mice, there was no significant change in the expression of MRP1 in Nrf2-/- model group. Conclusions: Mildly stable COPD mice may counteract the xenobiotic damage caused by cigarette smoke through up-regulating the expression of MRP1 protein, which may be associated with Nrf2 signaling activation.

Effect of maternal exposure to Curcumae Rhizoma during pregnancy on neurodevelopment and apoptosis mechanism in offspring / 中国中药杂志

Curcumae Rhizoma is a Chinese medicinal herb that is contraindicated during pregnancy. Cold-congelation and blood-stasis are corresponding syndromes to Curcumae Rhizoma. Whether syndrome-based treatment is associated with developmental neurotoxicity of Curcumae Rhizoma remains to be unclear. To verify the theory of traditional Chinese medicine of "syndrome-based treatment during pregnancy", the present study induced the mice blood stasis model by immersing mice in ice water. Pregnant C57 BL/6 wild type(WT) mice and pregnant Nrf2 knock out(KO) mice were randomly divided into control groups and Rhizoma Curcumae exposure groups. The mice were exposed to Rhizoma Curcumae during day 5 to day 18 after pregnancy. The neurodevelopment was examined to evaluate the differences of developmental neurotoxicity between normal and blood-stasis pregnant mice exposed to Rhizoma Curcumae. caspase-3 and caspase-9 activity in brain of the offspring were measured by colorimetric assays. Bcl-2 mRNA and protein expression in brain of the offspring were examined by Real-time RT-PCR and Western blot, respectively. According to the findings, C57 BL/6 mice exposed to Rhizoma Curcumae(10.0 g·kg~(-1)) had a longer positive occurring time of the surface righting reflex test of offspring and higher caspase-3 and caspase-9 activities in brain of offspring, compared with the normal control group, but with no significant change in those of blood-stasis pregnant mice offspring. However, mice exposed to Rhizoma Curcumae(10.0 g·kg~(-1)) showed no change in Bcl-2 gene expression and p38 MAPK phosphorylation in brain of the offspring. Nrf2 gene knockout using CRISPR/Cas9 resulted in a longer positive occurring time of the surface righting reflex test of offspring and higher caspase-3 and caspase-9 activities in brain of offspring. In conclusion, developmental neurotoxicity of the blood-stasis pregnant mice exposed to Rhizoma Curcumae was weaker than that of the normal pregnant mice. Nrf2 activation involved in the phenomenon of Rhizoma Curcumae of "syndrome-based treatment during pregnancy", but the upstream signal pathway mechanism value shall be further investigated.

MMP2 and MMP9 are Associated with Apical Periodontitis Progression and Might be Modulated by TLR2 and MyD88

Abstract The aim of this study was to evaluate the expression of MMP2 and MMP9 during apical periodontitis (AP) progression in TLR2 (TLR2 KO) and in MyD88 (MyD88 KO) knockout mice compared to wild type (WT) mice. AP was induced in mandibular first molars of TLR2 KO (n= 18), MyD88 KO (n= 18), and WT mice (n= 18). After 7, 21, and 42 days, the animals were euthanized and the jaws were dissected and subjected to histotechnical processing. Subsequent sections were stained by immunohistochemistry and evaluated for detection of MMP2 and MMP9. Statistical analysis of the semi-quantitative analysis of immunohistochemistry was performed using chi-square test (α = 0.05). In the initial periods of AP progression, an increased expression of MMP9 in the TLR2 KO and MyD88 KO mice was observed. In the final periods of AP progression, a reduction of MMP2 expression and an increase of MMP9 expression in the TLR2 KO mice were observed. MMP2 and MMP9 production was modulated for TLR2 and MyD88 during apical periodontitis progression.

Resumo O objetivo deste estudo foi avaliar a expressão de MMP2 e MMP9 durante a progressão da periodontite apical (AP) em camundongos knockout para TLR2 (TLR2 KO) e MyD88 (MyD88 KO) comparados aos camundongos wild type (WT). A AP foi induzida nos primeiros molares inferiores dos camundongos TLR2 KO (n = 18), MyD88 KO (n = 18) e WT (n = 18). Após 7, 21 e 42 dias, os animais foram eutanaziados e as mandíbulas foram dissecadas e submetidas a processamento histotécnico. As lâminas foram coradas por imuno-histoquímica e analisadas para a detecção de MMP2 e MMP9. A análise estatística semi-quantitativa da imuno-histoquímica foi realizada pelo teste qui-quadrado (α = 0,05). Nos períodos iniciais de progressão AP, foi observada uma expressão aumentada de MMP9 nos camundongos TLR2 KO e MyD88 KO. Nos períodos finais de progressão AP, observou-se uma redução da expressão de MMP2 e um aumento da expressão de MMP9 nos camundongos TLR2 KO. A produção de MMP2 e MMP9 foi modulada por TLR2 e MyD88 durante a progressão da periodontite apical.

Androgen receptor deficiency in monocytes/macrophages does not alter adiposity or glucose homeostasis in male mice / 亚洲男科学杂志(英文版)

Androgen deprivation in men leads to increased adiposity, but the mechanisms underlying androgen regulation of fat mass have not been fully defined. Androgen receptor (AR) is expressed in monocytes/macrophages, which are resident in key metabolic tissues and influence energy metabolism in surrounding cells. Male mice bearing a cell-specific knockout of the AR in monocytes/macrophages (M-ARKO) were generated to determine whether selective loss of androgen signaling in these cells would lead to altered body composition. Wild-type (WT) and M-ARKO mice (12-22 weeks of age, n = 12 per group) were maintained on a regular chow diet for 8 weeks and then switched to a high-fat diet for 8 additional weeks. At baseline and on both the regular chow and high-fat diets, no differences in lean mass or fat mass were observed between groups. Consistent with the absence of differential body weight or adiposity, no differences in food intake (3.0 ± 0.5 g per day for WT mice vs 2.8 ± 0.4 g per day for M-ARKO mice) or total energy expenditure (0.6 ± 0.1 Kcal h-1 for WT mice vs 0.5 ± 0.1 Kcal h-1 for M-ARKO mice) were evident between groups during high-fat feeding. Liver weight was greater in M-ARKO than that in WT mice (1.5 ± 0.1 g vs 1.3 ± 0.0 g, respectively, P = 0.02). Finally, M-ARKO mice did not exhibit impairments in glucose tolerance or insulin sensitivity relative to WT mice at any study time point. In aggregate, these findings suggest that AR signaling specifically in monocytes/macrophages does not contribute to the regulation of systemic energy balance, adiposity, or insulin sensitivity in male mice.

Can oliceridine (TRV130), an ideal novel µ receptor G protein pathway selective (µ-GPS) modulator, provide analgesia without opioid-related adverse reactions?

All drugs have both favorable therapeutic and untoward adverse effects. Conventional opioid analgesics possess both analgesia and adverse reactions, such as nausea, vomiting, and respiratory depression. The opioid ligand binds to µ opioid receptor and non-selectively activates two intracellular signaling pathways: the G protein pathway induce analgesia, while the β-arrestin pathway is responsible for the opioid-related adverse reactions. An ideal opioid should activate the G protein pathway while deactivating the β-arrestin pathway. Oliceridine (TRV130) has a novel characteristic mechanism on the action of the µ receptor G protein pathway selective (µ-GPS) modulation. Even though adverse reactions (ADRs) are significantly attenuated, while the analgesic effect is augmented, the some residual ADRs persist. Consequently, a G protein biased µ opioid ligand, oliceridine, improves the therapeutic index owing to increased analgesia with decreased adverse events. This review article provides a brief history, mechanism of action, pharmacokinetics, pharmacodynamics, and ADRs of oliceridine.

Androgen receptor deficiency in monocytes/macrophages does not alter adiposity or glucose homeostasis in male mice / 亚洲男科学杂志(英文版)

Androgen deprivation in men leads to increased adiposity, but the mechanisms underlying androgen regulation of fat mass have not been fully defined. Androgen receptor (AR) is expressed in monocytes/macrophages, which are resident in key metabolic tissues and influence energy metabolism in surrounding cells. Male mice bearing a cell-specific knockout of the AR in monocytes/macrophages (M-ARKO) were generated to determine whether selective loss of androgen signaling in these cells would lead to altered body composition. Wild-type (WT) and M-ARKO mice (12-22 weeks of age, n = 12 per group) were maintained on a regular chow diet for 8 weeks and then switched to a high-fat diet for 8 additional weeks. At baseline and on both the regular chow and high-fat diets, no differences in lean mass or fat mass were observed between groups. Consistent with the absence of differential body weight or adiposity, no differences in food intake (3.0 ± 0.5 g per day for WT mice vs 2.8 ± 0.4 g per day for M-ARKO mice) or total energy expenditure (0.6 ± 0.1 Kcal h-1 for WT mice vs 0.5 ± 0.1 Kcal h-1 for M-ARKO mice) were evident between groups during high-fat feeding. Liver weight was greater in M-ARKO than that in WT mice (1.5 ± 0.1 g vs 1.3 ± 0.0 g, respectively, P = 0.02). Finally, M-ARKO mice did not exhibit impairments in glucose tolerance or insulin sensitivity relative to WT mice at any study time point. In aggregate, these findings suggest that AR signaling specifically in monocytes/macrophages does not contribute to the regulation of systemic energy balance, adiposity, or insulin sensitivity in male mice.