Migrasomes are migration-dependent membrane-bound vesicular structures that contain cellular contents and small vesicles. Migrasomes grow on the tips or intersections of the retraction fibers after cells migrate away. The process of releasing migrasomes into the extracellular space is named as “migracytosis”. After releasing, they can be taken up by the surrounding cells, or rupture and further release their contents into the extracellular environment. Physiologically, migrasomes provide regional cues for organ morphogenesis during zebrafish gastrulation and discard the damaged mitochondria in response to mild mitochondrial stresses. Pathologically, migrasomes are released from podocyte during early podocyte stress and/or damage, from platelets after infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from microglia/macrophages of the ischemic brain, and from tumor necrosis factor α (TNFα)-activated endothelial cells (ECs); thus, this newly discovered extracellular vesicle is involved in all these pathological processes. Moreover, migrasomes can modulate the proliferation of cancer cell via lateral transferring mRNA and protein. In this review, we will summarize the biogenesis, release, uptake, and rupture of migrasomes and discuss its biological roles in development, redox signalling, innate immunity and COVID-19, cardio-cerebrovascular diseases, renal diseases, and cancer biology, all of these highlight the importance of migrasomes in modulating body homeostasis and diseases.

目的 探讨FPR2在补阳还五汤减轻大鼠脑缺血再灌注中作用.方法 运用线栓法建立大脑中动脉栓塞(MCAO)模型.实验动物随机分为假手术组、模型组、补阳还五汤组和补阳还五汤联合FPR2抑制剂(Boc-2)组.补阳还五汤剂量为每次16 g/kg,每天灌胃2次,术前30 min腹腔注射Boc-2 (0.4 mg/kg),缺血后6h,24h,2d,3d,7d,运用Western blot检测FPR2蛋白表达和ELISA检测FPR2内源性配体LXA4和RvD1蛋白表达;缺血后24h进行平衡木和抓力评分,以及测定脑梗死面积.结果 与模型组比,补阳还五汤组FPR2和RvD1在缺血后24h达到高峰(P＜0.05);据此,选择缺血后24h为后续实验时间点,发现补阳还五汤能改善大鼠平衡功能、抓力和减小脑梗死面积(P＜0.01,P＜0.05);但联合使用Boc-2后,大鼠行为学功能下降和脑梗死面积增加(P＜0.01,P＜0.05).结论 补阳还五汤可能通过增强FPR2作用减轻大鼠脑缺血再灌注神经功能损伤、减少脑梗死面积,从而发挥保护作用.

Sex is a science of cutting edge but bathed in mystery. Coitus or sexual intercourse, which is at the core of sexual activities, requires healthy and functioning vessels to supply the pelvic region, thus contributing to clitoris erection and vaginal lubrication in female and penile erection in male. It is well known that nitric oxide (NO) is the main gas mediator of penile and clitoris erection. In addition, the lightest and diffusible gas molecule hydrogen (H2) has been shown to improve erectile dysfunction (ED), testis injuries, sperm motility in male, preserve ovarian function, protect against uterine inflammation, preeclampsia, and breast cancer in female. Mechanistically, H2 has strong abilities to attenuate excessive oxidative stress by selectively reducing cytotoxic oxygen radicals, modulate immunity and inflammation, and inhibit injuries-induced cell death. Therefore, H2 is a novel bioactive gas molecule involved in modulating sexual organs homeostasis.

PURPOSE:Skeletal muscle has a major influence on whole-body metabolic homeostasis. In the present study, we aimed to determine the metabolic effects of the β3 adrenergic receptor agonist CL316243 (CL) in the skeletal muscle of high-fat diet-fed rats.METHODS:Sprague-Dawley rats were randomly allocated to three groups, which were fed a control diet (C) or a high-fat diet (HF), and half of the latter were administered 1 mg/kg CL by gavage once weekly (HF+CL), for 12 weeks. At the end of this period, the serum lipid profile and glucose tolerance of the rats were evaluated. In addition, the phosphorylation and protein and mRNA expression of AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor γ coactivator (PGC)-1α, and carnitine palmitoyl transferase (CPT)-1b in skeletal muscle were measured by Western blot analysis and qPCR. The direct effects of CL on the phosphorylation (p-) and expression of AMPK, PGC-1α, and CPT-1b were also evaluated by Western blotting and immunofluorescence in L6 myotubes.RESULTS:CL administration ameliorated the abnormal lipid profile and glucose tolerance of the high-fat diet-fed rats. In addition, the expression of p-AMPK, PGC-1α, and CPT-1b in the soleus muscle was significantly increased by CL. CL (1 µM) also increased the protein expression of p-AMPK, PGC-1α, and CPT-1b in L6 myotubes. However, the effect of CL on PGC-1α protein expression was blocked by the AMPK antagonist compound C, which suggests that CL increases PGC-1α protein expression via AMPK.CONCLUSION:Activation of the β3 adrenergic receptor in skeletal muscle ameliorates the metabolic abnormalities of high-fat diet-fed rats, at least in part via activation of the AMPK/PGC-1α pathway.

*These authors contributed equally to this work Purpose: Skeletal muscle has a major influence on whole-body metabolic homeostasis. In the present study, we aimed to determine the metabolic effects of the β3 adrenergic receptor agonist CL316243 (CL) in the skeletal muscle of high-fat diet-fed rats. Methods: Sprague-Dawley rats were randomly allocated to three groups, which were fed a control diet (C) or a high-fat diet (HF), and half of the latter were administered 1 mg/kg CL by gavage once weekly (HF+CL), for 12 weeks. At the end of this period, the serum lipid profile and glucose tolerance of the rats were evaluated. In addition, the phosphorylation and protein and mRNA expression of AMP-activated protein kinase (AMPK), peroxisome proliferator-activated receptor γ coactivator (PGC)-1α, and carnitine palmitoyl transferase (CPT)-1b in skeletal muscle were measured by Western blot analysis and qPCR. The direct effects of CL on the phosphorylation (p-) and expression of AMPK, PGC-1α, and CPT-1b were also evaluated by Western blotting and immunofluorescence in L6 myotubes. Results: CL administration ameliorated the abnormal lipid profile and glucose tolerance of the high-fat diet-fed rats. In addition, the expression of p-AMPK, PGC-1α, and CPT-1b in the soleus muscle was significantly increased by CL. CL (1 μM) also increased the protein expression of p-AMPK, PGC-1α, and CPT-1b in L6 myotubes. However, the effect of CL on PGC-1α protein expression was blocked by the AMPK antagonist compound C, which suggests that CL increases PGC-1α protein expression via AMPK. Conclusion: Activation of the β3 adrenergic receptor in skeletal muscle ameliorates the metabolic abnormalities of high-fat diet-fed rats, at least in part via activation of the AMPK/ PGC-1α pathway.

Objective: Synaptic plasticity is critical for neurorehabilitation after focal cerebral ischemia. Connexin 43 (Cx43), the main component of the gap junction, has been shown to be pivotal for synaptic plasticity. The objective of this study was to investigate the role of the Cx43 inhibitor (Gap26) and gap junction modifier (GAP-134) in neurorehabilitation and to study their contribution to synaptic plasticity after focal ischemia. Methods: Time course expression of both total and phosphorylated Cx43 (p-Cx43) were detected by western blotting at 3, 7, and 14 d after focal ischemia. Gap26 and GAP-134 were administered starting from 3 d post focal ischemia. Neurological performances were evaluated by balance beam walking test and Y-maze test at 1, 3, and 7 d. Golgi staining and transmission electron microscope (TEM) detection were conducted at 7 d for observing dendritic spine numbers and synaptic ultrastructure, respectively. Immunofluorescent staining was used at 7 d for detection of synaptic plasticity markers, including synaptophysin (SYN) and growth-associated protein-43 (GAP-43). Results: Expression levels of both total Cx43 and p-Cx43 were increased after focal cerebral ischemia, peaking at 7 d. Compared with the MCAO group, Gap26 worsened the neurological behavior and decreased the dendritic spine number while GAP-134 improved the neurobehavior and increased the number of dendritic spines. Moreover, Gap26 further destroyed the synaptic structure, concomitant with downregulated SYN and GAP-43, whereas GAP-134 alleviated synaptic destruction and upregulated SYN and GAP-43. Conclusion: These findings suggested that Cx43 or the gap junction was involved in synaptic plasticity, thereby promoting neural recovery after ischemic stroke. Treatments enhancing gap junctions may be potential promising therapeutic measures for neurorehabilitation after ischemic stroke.

目的:探讨补阳还五汤在脑缺血再灌注大鼠恢复期提高突触可塑性的机制研究.方法:建立大脑中动脉栓塞(MCAO)大鼠模型,随机分为假手术组、模型组、补阳还五汤组、补阳还五汤联合缝隙连接蛋白43(Cx43)抑制剂(Gap26)组,补阳还五汤每天灌胃2次(16 g·kg-1),Gap26于术后第3天腹腔注射,每天1次(25μg·kg-1);7 d后取材,采用透射电镜观察缺血侧海马突触和缝隙连接超微结构的改变,运用蛋白免疫印迹法(Western blot)和免疫荧光检测缺血侧海马突触素(SYN),生长相关蛋白-43(GAP-43)的表达.结果:电镜下观察到假手术组突触结构完整、清晰,突触数量多,缝隙连接结构清晰;模型组缺血侧海马突触结构溶解,突触数量减少,缝隙连接消失,存在较大间隙,与假手术组比较,SYN,GAP-43的表达明显增高(P＜0.05,P＜0.01);补阳还五汤组缺血侧海马突触结构较清晰,突触数量增多,缝隙连接结构较完整,与模型组比较,SYN,GAP-43的表达明显增强(P＜0.05,P＜0.01);而联合使用Gap26后缺血侧海马突触数量较补阳还五汤组减少,仅可见少量结构完整的缝隙连接,补阳还五汤增强SYN,GAP-.43的作用被明显抑制(P＜0.05,P＜0.01).结论:补阳还五汤可提高脑缺血再灌注恢复期缺血侧海马突触可塑性,其机制可能与增加Cx43的表达促进对SYN,GAP-43的干预有关.

目的:探讨星形胶质细胞缝隙连接蛋白43(connexin43,Cx43)在补阳还五汤促进大鼠脑缺血再灌注后修复中的作用。方法:运用线栓法建立大脑中动脉栓塞(MCAO)模型,将造模后的大鼠随机分为模型组,补阳还五汤(BYHWD)组和补阳还五汤联合Cx43抑制剂(BYHWD+Gap26)组,另设假手术对照组。在术后第1、3、7天对各组大鼠进行横木行走测试(BWT)和自发交替反应测试(SAB test);第7天取各组大鼠脑组织,采用高尔基染色法观察海马CA1、CA3和DG区神经元基、顶树突的长度,节点数,终末分支数以及树突棘密度的改变;采用免疫荧光法以CD31表达量计算海马CA1、CA3和DG区的微血管密度(MVD)。结果:与模型组比较,BYHWD组大鼠衡木行走测试评分降低(P<0.05,P<0.01),自发交替反应测试相对交替率升高(P<0.05,P<0.01);神经元树突长度、节点数、终末分支数以及树突棘密度大体呈增长的趋势;海马CA1、CA3和DG区MVD表达水平均升高(P<0.05,P<0.01)。相较于BYHWD组,BYHWD+Gap26组第7天大鼠横木行走测试评分更高(P<0.05),自发交替反应测试相对交替率更低(P<0.05);神经元树突长度、节点数、终末分支数以及树突棘密度大体呈下降的趋势;海马CA1、CA3和DG区MVD表达水平均显著降低(P<0.01)。结论:补阳还五汤可通过星形胶质细胞Cx43促进神经元修复和血管再生,从而改善脑缺血再灌后的神经功能情况。

目的:探讨星形胶质细胞缝隙连接蛋白43(connexin43,Cx43)在补阳还五汤促进大鼠脑缺血再灌注后修复中的作用.方法:运用线栓法建立大脑中动脉栓塞(MCAO)模型,将造模后的大鼠随机分为模型组,补阳还五汤(BYHWD)组和补阳还五汤联合Cx43抑制剂(BYHWD+ Gap26)组,另设假手术对照组.在术后第1、3、7天对各组大鼠进行横木行走测试(BWT)和自发交替反应测试(SAB test);第7天取各组大鼠脑组织,采用高尔基染色法观察海马CA1、CA3和DG区神经元基、顶树突的长度,节点数,终末分支数以及树突棘密度的改变;采用免疫荧光法以CD31表达量计算海马CA1、CA3和DG区的微血管密度(MVD).结果:与模型组比较,BYHWD组大鼠衡木行走测试评分降低(P ＜0.05,P＜0.01),自发交替反应测试相对交替率升高(P＜0.05,P＜0.01);神经元树突长度、节点数、终末分支数以及树突棘密度大体呈增长的趋势;海马CA1、CA3和DG区MVD表达水平均升高(P＜0.05,P＜0.01).相较于BYHWD组,BYHWD+ Gap26组第7天大鼠横木行走测试评分更高(P＜0.05),自发交替反应测试相对交替率更低(P＜0.05);神经元树突长度、节点数、终末分支数以及树突棘密度大体呈下降的趋势;海马CA1、CA3和DG区MVD表达水平均显著降低(P＜0.01).结论:补阳还五汤可通过星形胶质细胞Cx43促进神经元修复和血管再生,从而改善脑缺血再灌后的神经功能情况.

目的 观察不同状态下培养的乳鼠心肌细胞上清液对大鼠骨髓间充质干细胞(MSCs)心肌动作电位相关离子通道基因表达的影响.方法 分别取大鼠骨髓MSCs和乳鼠心肌细胞进行体外培养.实验分为MSCs组、5-氮胞苷(5-Aza)诱导组、正常诱导组、缺氧诱导组和心肌细胞组,MSCs组为正常培养的MSCs,5-Aza诱导组为心肌定向诱导剂5-Aza干预的MSCs,正常诱导组为正常心肌细胞上清液干预的MSCs,缺氧诱导组为缺氧状态下的心肌细胞上清液干预的MSCs,心肌细胞组为不予干预的正常心肌细胞.采用RT-PCR法检测各组细胞膜上钠通道(SCN2a)、钾通道(Kv2.1、Kv1.4、Kir1.1、EAG1)和钙通道基因(CCHL2α)mRNA表达的变化.结果 MSCs组细胞膜上表达与心肌动作电位相关的离子通道基因.与MSCs组相比,其余各组SCN2a、Kv2.1、Kir1.1、EAG1、CCHL2αmRNA表达均升高(P<0.05或<0.01),且缺氧诱导组、心肌细胞组Kv1.4 mRNA表达升高(P<0.05或<0.01),但5-Aza诱导组、正常诱导组、缺氧诱导组各种离子通道基因表达两两比较差异无统计学意义.结论 乳鼠正常心肌细胞上清液和缺氧心肌细胞上清液可诱导大鼠MSCs心肌动作电位相关离子通道基因表达增加,心肌微环境有利于大鼠MSCs发生心肌电生理变化.

Objective:To study the anti-atherosclerosis (AS) mechanism of resveratrol (RES) by regulating store-operated calcium channel (SOCC) induced by transient receptor potential channel protein 1 (TRPC1) and stromal interaction molecule 1 (STIM1).Method:Atherosclerosis model was established with female C57BL/6J mice that were fed with high-fat diet after ovariectomy,and then the mice were divided into control group,model group,low-dose resveratrol group,medium-dose resveratrol group,high-dose resveratrol group (50,100,150 mg·kg-1) and estradiol (E2) group (0.3 mg·kg-1).After 16 weeks,serum was collected,and the lipid level and Ca2+ concentration were detected.The morphological changes were measured by oil red O staining.The protein expressions of TRPC1,STIM1 and endothelial nitric oxide synthase (eNOS) were detected by Western blot and Real-time PCR.The activity of eNOS was measured by enzyme-linked immunosorbent assay (ELISA).Result:Compared with model group,total cholesterol (TC),triglyceride (TG),low density lipoprotein (LDL) were decreased,while the high density lipoprotein (HDL) was increased in supplementary high-dose resveratrol or estrogen groups (P ＜ 0.05).The pathological changes in the model group were obvious compared with control group,after supplementation with RES or estrogen,the pathological changes were significantly reduced in thoracic aorta.Western blot and Real-time PCR results showed that in model group,the protein expressions of TRPC1 and STIM1 were increased,while the protein expression of eNOS was decreased (P ＜ 0.05);in supplementary high-dose resveratrol or estrogen group,these changes were reversed (P ＜ 0.05).Conclusion:RES could down-regulate TRPC1 and STIM1 gene and protein expressions,inhibit SOCC-mediated Ca2+ influx,increase eNOS expression and resist atherosclerosis.

目的 研究白藜芦醇(Resveratrol,RSV)通过瞬时受体电位C1通道(transient receptor potential canonical-channel 1,TRPC1)来抑制血管平滑肌细胞(Vascular smooth muscle cells,VSMCs)增殖的机制.方法 采用血管环培养法观察RSV对胎牛血清(fetal calf serum,FBS)诱导的血管壁损伤的影响;MTT法检测RSV对VSMCs增殖的影响;qPCR法检测RSV对TRPC1 mRNA表达的影响;Western Blot法检测RSV对TRPC1蛋白表达的影响.结果 与模型组比较,RSV在20～200μmol·L-1范围内,可改善高血清诱导的血管壁损伤和管壁增厚现象;RSV可降低FBS诱导的VSMCs增殖(P<0.01),以200μmol·L-1浓度最为显著,此作用可被TRPC1抑制剂SKF96365所阻断;RSV各浓度组的TRPC1 mRNA及蛋白相对表达量显著降低(P<0.01).结论 RSV抑制VSMCs的增殖可能通过影响TRPC1靶点来实现.

目的:探讨缝隙连接蛋白43(eonnexin43,Cx43)在补阳还五汤抗脑缺血损伤中的作用.方法:采用大脑中动脉栓塞制备脑缺血模型,缺血2h后拔出线栓进行再灌注.补阳还五汤灌胃,24 h后取材通过免疫蛋白印迹检测Cx43,苏木精-伊红染色(hematoxyli-eosin,HE)观察脑组织神经元损伤,以及相关试剂盒检测丙二醛(malondialdehyde,MDA)、一氧化氮(nitric oxide,NO)和超氧化物岐化酶(superoxide dismutase,SOD).结果:免疫蛋白印迹结果表明,补阳还五汤可下调缺血后Cx43的表达.另外,联合使用Cx43激动剂(GAP-134)后神经元损伤较补阳还五汤组加重,提示Cx43参与了补阳还五汤的抗脑缺血损伤.进一步通过试剂盒检测发现,同模型组相比,补阳还五汤可降低脑缺血后NO和MDA的含量,上调SOD含量.而使用Cx43激动剂后,补阳还五汤对NO的下调和对SOD的上调作用均被抑制.结论:这些结果提示Cx43可能是通过减少NO以及增加SOD介导补阳还五汤对脑缺血损伤的保护作用.

Recent studies have demonstrated that Buyang Huanwu Decoction (BYHWD) decreased glutamate levels subsequent to cerebral ischemia. Glutamate transporter-1 (GLT-1) and glutamine synthetase (GS), which are located in astrocytes, mainly contribute to glutamate transportation, thus reducing glutamate concentration. BYHWD has previously been demonstrated to upregulate GLT-1 and GS following ischemia in vivo. However, whether BYHWD can directly influence astrocytic GLT-1/GS levels remains unknown. In the present study, the effect of BYHWD containing serum (BYHWD-CS) on GLT-1/GS levels in astrocytes following oxygen-glucose deprivation/reoxygenation (OGD/R) was investigated. The results revealed that BYHWD-CS enhanced the expression levels of GLT-1 and GS in cultured astrocytes, which reduced glutamate concentration in the culture medium. Meanwhile, increased p38 mitogen-activated protein kinase (p38 MAPK) was phosphorylated (activation form) by BYHWD-CS in cultured astrocytes, and the specific p38 inhibitor SB203580 blocked the increase of GLT-1/GS accompanied by decreased cell viability. Furthermore, SB203580 suppressed the effect of BYHWD-CS on the level of glial fibrillary acidic protein (an astrocytic marker), thus confirming that astrocytes are directly involved in the protective role of BYHWD after OGD/R. These findings suggest that BYHWD upregulates GLT-1 and GS via p38 MAPK activation, and protects cultured astrocytes from death caused by OGD/R (typical in vitro model), which complemented the role of astrocytes in the protective effect of BYHWD.

目的 探讨补阳还五汤对大脑中动脉栓塞后不同时间大鼠脑内微小核糖核酸(miRNA)表达的影响.方法 建立大脑中动脉栓塞的脑缺血模型,芯片方法观察miRNA的表达差异,RT-PCR进行验证.结果 与3天模型组相比,补阳还五汤组miRNA有2个上调,1个下调;与15天模型组相比,补阳还五汤组miRNA有5个上调,4个下调.随后,选取3个miRNA使用RT-PCR验证各组大鼠脑内miRNA差异表达,同芯片结果相符.结论 补阳还五汤对脑缺血后3天和15天miRNA均能产生影响,提示补阳还五汤对脑缺血急性期和修复期都具有治疗作用.另外,相比较而言,补阳还五汤对脑缺血后15天miRNA影响的数量更多,可能其促进修复的作用更具有优势.

Objective:To explore the mechanisms of Buyang Huanwu Decoction (BYHWD) influencing connexin 43 (Cx43) expression after focal cerebral ischemia in rats.Methods:Middle cerebral artery occlusion(MCAO) and reperfusion model was established by Longa method.One day after treatment with BYHWD,ELISA was used to detect levels of inflammatory cytokines (interleukin-1,IL-1β,interleukin-10,IL-10).Cx43 expression was compared by immunohistochemical staining after treatment with NF-κB agonist,and HE staining was used to observe cell morphological changes.Seven days after treatment with BYHWD,immunohistochemical staining was used to investigate the expression of bFGF and Cx43,and HE staining was used to observe cell morphology changes.Results:Compared with ischemia group,IL-1 β was decreased whereas IL-10 was increased by BYHWD on day 1.The downregulation of Cx43 expression in hippocampus CA1 by BYHWD was inhibited by combination treatment with NF -κB agonist on day 1 (P ＜ 0.05) and neuronal damage was aggravated in combination group.On day 7 afer focal ischemia,bFGF and Cx43 expressions were higher in BYHWD group than that in ischemia group (P ＜ 0.05).Cx43 expression was significantly decreased after combination of bFGF neutralizing antibody with BYHWD (P ＜ 0.05),meanwhile neuronal damage was aggravated in combination group.Conclusion:The effects of BYHWD on Cx43 may be regulated by NFκB or bFGF and related with its curative effects of alleviating injury and facilitating functional restoration after focal cerebral ischemia and reperfusion.

ObjectiveTo investigate the effects ofErxianDecoction(ED)on the expression levels of thermogenesis- related proteins in interscapular brown adipose tissue of ovariectomized rats.MethodsForty female SD rats were randomly divided into sham operation group,ovariectomized group,estrogen group,and low- and high-dose ED groups,which were treated with normal saline,estrogen,ED respectively for 14 weeks. Serum lipids levels were tested. By using HE staining,the morphological features of interscapular brown adipose tissue of rats in each group were observed. The protein and mRNA expression levels of uncoupling protein-1(UCP-1)and peroxisome proliferator- activated receptor co-activator 1α(PGC-1α)in rat interscapular brown adipose tissue of each group were determined by Western blot and qPCR.ResultsCompared with sham operation group,rats in ovariectomized group had high body weight,intra-abdominal fat weight and serum cholesterol level. ED treatment was able to decrease body weight, intra-abdominal fat weight and serum cholesterol level in ovariectomized rats. The protein and mRNA levels of UCP-1 and PGC-1αin interscapular brown adipose tissue of ovariectomized rats were significantly reduced,and ED treatment could increase the protein and mRNA levels of UCP-1 and PGC-1αin ovariectomized rats.ConclusionED treatment is effective on decreasing the body weight,interscapular fat weight and serum total cholesterol level in ovariectomized rats. The mechanism is probably associated with the up-regulation of protein and mRNA expression levels of thermogenesis-related proteins UCP-1 and PGC-1αin interscapular brown adipose tissue of ovariectomized rats.

Background: Perivascular adipose tissue (PVAT) can regulate vascular homeostasis by secreting various adipokines. This study investigated the effects of PVAT browning on its endocrine function. Methods: In the first section of our study, male Sprague-Dawley rats were randomly divided into cold exposure (8°C) and 24°C acclimation groups. After cold exposure for 7 days, interscapular brown adipose tissue (iBAT), subcutaneous white adipose tissue, thoracic aortic PVAT, and abdominal aortic PVAT (aPVAT) were harvested for histological and brown marker gene expression analysis. In the second part, male rats were fed a high fat diet (HFD) for 10 weeks. In the 11th week, the rats were treated with or without cold exposure. After 14-day cold exposure, aPVAT was collected for histological, gene, and protein expression analysis. Results: Cold exposure had a browning effect on aPVAT by increasing UCP-1 and PGC-1α expression levels. After HFD feeding for 10 weeks, 14-day cold exposure was still able to induce aPVAT browning. Compared with thermoneutrality acclimation rats, TNF-α, IL-6, and p-p65 expression levels were significantly lower in aPVAT from HFD-fed rats with cold exposure. In contrast, p-AMPK expression levels were increased in aPVAT from HFD-fed rats with cold exposure. Conclusions: Our study demonstrated that browning of aPVAT in HFD-fed rats lowered the pro-inflammatory adipokine expression levels and activated AMPK.

目的 观察补阳还五汤对全脑缺血再灌注后大鼠皮层神经元NR2B mRNA表达的影响,探讨补阳还五汤抗全脑缺血后再灌注损伤的分子机制.方法 SD大鼠随机分为3组,即正常组、模型组和药物组(补阳还五汤预干预),模型组和药物组再分别设置2、24h两个观察时间点.连续给药5d后,采用改良的Pulsinelli四血管闭塞法复制全脑缺血大鼠模型,再灌后2、24 h采用RT-PCR技术检测各组大鼠皮质神经细胞NMDA受体(N-methyl-D-aspartic acid receptor)亚单位NR2B的mRNA表达水平.结果 与正常组比较,模型组2h、24h等2个不同时间点,NR2B的mRNA表达水平显著升高(P＜0.05);与模型组比较,药物组再灌注后2h、24h等2个不同时间点,NR2B mRNA表达水平显著降低(P＜0.01).结论 补阳还五汤干预对全脑缺血再灌注模型大鼠大脑皮层神经元NR2B mRNA表达水平的增高有调低作用.

The neuroprotective role of Buyang Huanwu decoction (BYHWD) in focal ischemia is associated with decreasing glutamate concentration. However, the mechanisms are not fully understood. The present study aimed to explore whether glutamate transporter-1 (GLT-1) and glutamine synthetase (GS) participated in the decreased level of glutamate and whether pituitary adenylate cyclase-activating polypeptide-38 (PACAP-38) was involved in this process. BYHWD was found to significantly upregulate the expression of GLT-1 and GS in the hippocampal CA1 area compared to the ischemia group, with the difference on day 3 being most significant. BYHWD increased the level of PACAP-38, and PACAP-(6-38) (PACAP receptor antagonist) significantly attenuated the effect of BYHWD on GLT-1 and GS, suggesting that PACAP-38 was involved in the upregulation of GLT-1 and GS induced by BYHWD. In addition, as GLT-1 and GS are mainly located in astrocytes, the changes of astrocytes were detected by glial fibrillary acidic protein (GFAP; an astrocytic marker) immunostaining. The results showed that BYHWD inhibited the expression of GFAP compared with the ischemia group, however, co-administration with PACAP-(6-38), which inhibited the effect of BYHWD on GLT-1 and GS in astrocytes, attenuated this effect, indicating that astrocytes participated in the protective role of BYHWD following focal ischemia. These results provided the evidence for the first time that not only neurons but also astrocytes contribute to the protective role of BYHWD, which opposes previous studies and may be a starting point for traditional medicine.