High-fat diet (HFD)-induced microglial activation contributes to hypothalamic inflammation and obesity, but the mechanisms linking microglia to structural changes remain unclear. This study explored the role of microglia in impairing hypothalamic synaptic plasticity in diet-induced obesity mice and evaluated the therapeutic potential of semaglutide (Sema) and minocycline (MI). Six-week-old C57BL/6J mice were divided into low-fat diet and HFD groups. At week 30, the HFD-fed mice were treated daily with Sema or MI for six weeks. Confocal microscopy assessed hypothalamic dendritic spines, synaptic organization, and microglia-synapse interactions. We also analyzed microglial morphology, CD68/CD11b colocalization with Iba-1, synaptic marker expression, and phagocytosis-related pathways (C1q, C3, CD11b). BV2 microglia were used to examine the direct effects of Sema or MI on microglia and validate the in vivo findings. HFD feeding induced microglial activation, as indicated by increased colocalization of CD68 or synaptophysin and CD11b with Iba-1, along with elevated C1q, C3, and CD11b expression, signaling enhanced synaptic phagocytosis. This was accompanied by reduced hypothalamic dendritic spines, decreased synaptic marker expression, and disrupted excitatory/inhibitory synaptic organization in the melanocortin system, as well as impaired glucose metabolism, disrupted leptin-ghrelin balance, and increased food intake and body weight. Sema and MI treatments reversed the pathological changes of microglial activation and restored hypothalamic synaptic structure, although their effects on synaptic organization and metabolic outcomes differed. Our findings highlight the key role of microglial activation in hypothalamic synaptic impairment in diet-induced obesity models, with Sema and MI possibly offering distinct therapeutic pathways to mitigate these impairments.

目的:基于Toll样受体4(TLR4)信号通路,探索利拉鲁肽抑制脂多糖(LPS)诱导的原代小胶质细胞炎症反应的机制.方法:选取出生1 d SPF级SD大鼠,分离培养原代小胶质细胞,利用免疫荧光染色特异性蛋白Iba-1检测其纯度,将细胞随机分为5组:空白对照(N)组、脂多糖(LPS)组、利拉鲁肽(Li)组、脂多糖+利拉鲁肽(LL)组及脂多糖+TLR4抑制剂(LT)组.LPS组用LPS干预24 h,Li组用利拉鲁肽干预24 h,LL组用LPS干预24 h+利拉鲁肽干预24 h,LT组用LPS干预24 h+TAK-242干预24 h.用流式细胞仪测定各组细胞总凋亡率,以蛋白质免疫印迹法(Western blotting)检测IL-1β、TNF-α、TLR4、MyD88、TRAF6蛋白在各组中的表达情况.结果:与N组比较,LPS组细胞总凋亡率上升(P＜0.05);与LPS组比较,LL组和LT组细胞的总凋亡率呈下降趋势(P＜0.05).与N组比较,LPS组IL-1β、TNF-α、TLR4、MyD88、TRAF6蛋白相对表达量增高(P＜0.05);与LPS组比较,LL组和LT组IL-1β、TNF-α、TLR4、MyD88、TRAF6蛋白表达呈下降趋势(P＜0.05).结论:利拉鲁肽可能通过下调TLR4信号通路中关键因子表达抑制LPS诱导的原代小胶质细胞炎症反应,并改善细胞凋亡,保护神经系统.

1 临床资料 患者,女,15岁,因"口渴多饮、双下肢乏力6月余"于2019年7月首次入院.2019年1月出现口渴多饮、双下肢乏力伴消瘦,诊断为"糖尿病",曾尝试使用磺脲类药物降糖,血糖控制欠佳,后期改胰岛素降糖,2019年6月自行停用胰岛素,逐渐出现头部双颞侧放电样痛.既往有慢性胃炎病史.家族史:母亲否认糖尿病史,奶奶有糖尿病史,父亲智能低下,因"脑卒中"已去世.

目的:探索脂多糖(LPS)干预原代小胶质细胞时间与表达诱导型一氧化氮合酶(iNOS)时效关系的研究,为小胶质细胞建立炎症模型提供依据.方法:把提纯后的原代小胶质细胞随机分为实验组和对照组.实验组分别用LPS干预6h、12h、24 h和48 h,对照组为LPS未干预组,即LPS干预0h.用免疫荧光和蛋白质免疫印迹法(Western blot)分别检测iNOS在小胶质细胞中的表达.结果:实验组LPS干预小胶质细胞6h、12h、24h和48h后iNOS的表达量增高,其中12h,24h,48h与对照组0h比较,差异有统计学意义(P＜0.01),且在24h前逐渐升高,24h达峰值,24h后逐渐下降.结论:iNOS表达量与LPS干预原代小胶质细胞的时间有关,LPS干预24h时iNOS表达最高,LPS干预24h为理想的小胶质细胞炎症模型.

Objective:To investigate the effects of liraglutide and aerobic exercise on morphology and apoptosis of arcuate nucleus (ARC)neurons in obese rats.Methods:Obese rats were established by a high-fat diet and randomly divided into high-fat diet+saline (HS) group,high-fat diet+liraglutide (HL) group,and high-fat diet + aerobic exercise (HE) group.Untreated healthy rats served as the normal control(NC) group.The HL group rats received subcutaneous injection of liraglutide 200 μg/(kg· d).The NC group and HS group rats were given subcutaneous injection of the same volume of normal saline.Rats in the HE group were subjected to weightless swimming training for 60 min once day.After 6 weeks of intervention,the morphological structure of neurons in ARC of hypothalamus was observed by Nissl staining.The expression levels of Bax and Bcl-2 were determined by immunofluorescence.The apoptotic rate was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.Results:Compared with the NC group,ARC neurons were occasionally pyknotic and decreased in number in the hippocampus of the HS rats,and exhibited disordered arrangement,while milder neuronal injury and more cells were observed in the rats of the HL group and HE group.Compared with the HS group,liraglutide treatment or aerobic exercise decreased the number of TUNEL-positive cells and the expression of Bax concomitantly with upregulated Bcl-2 (P＜0.01).The changes were more marked in the HL group.Conclusion:Liraglutide could alleviate neuronal damages of the ARC by inhibiting apoptosis in obese rats,and its effect was superior to that of aerobic exercise.

目的:探讨沙格列汀联合预混胰岛素治疗2型糖尿病的效果及其对血糖波动的影响.方法:选取2017年5月至2018年3月广西医科大学第一附属医院老年内分泌科收治的100例2型糖尿病患者,按随机数字表法分为观察组和对照组,每组50例,其中对照组给予门冬胰岛素30注射液治疗,观察组给予沙格列汀联合门冬胰岛素30注射液治疗.检测两组患者治疗前、后空腹血糖(FBG)、餐后2h血糖(2 hPG)、三酰甘油(TG)、胆固醇(TC)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)、糖化血红蛋白(HbA1c)、空腹胰岛素(Fins)、空腹血清C肽(FCP)及餐后2 hC肽(2 hCP)水平,计算胰岛β细胞功能指数(HO-MA-β)和胰岛素抵抗指数(HOMA-IR),并应用动态血糖监测系统监测24 h平均血糖水平.结果:治疗后,观察组HbA1c、FBG、2 hBG、LDL-C、HOMA-IR显著低于治疗前及对照组,2 hCP显著高于治疗前及对照组(均P＜0.05).治疗后,观察组TG水平显著低于治疗前,HDL-C、FCP、HOMA-β水平显著高于治疗前(P＜0.05),但与对照组比较差异无统计学意义(P＞0.05).观察组午餐前1h、晚餐前1h、早餐后3h、午餐后3h、晚餐后3h平均血糖水平、日内血糖平均波动幅度及日间血糖平均绝对差均显著低于对照组(均P＜0.05).结论:沙格列汀联合预混胰岛素对2型糖尿病患者的血糖控制效果更佳,并能降低血糖波动.

Objective:To investigate the effect of ghrelin on the autophagy of islet β cells under high glucose and high lipid (HGHL) environment.Methods:Islet NIT-1cells were cultured and divided into four groups:control group(glucose 5.6 mmol/L) ,ghrelin group(glucose 5.6mmol/L+100nmol/L ghrelin) ,HGHL group(glucose 33.3mmol/L+0.5mmol/L palmitic acid) ,HGHL+ghrelin group (glucose 33.3mmol/L+0.5mmol/L PA+100nmol/L ghrelin).The cell counting kit (CCK-8) was used to assess the NIT-1cells viability. Monodansylcadaverine(MDC) staining and microtubule-associated protein 1light chain 3 (LC3) B immunofluorescence staining were employed to detect the autophagy.Western blotting was performed to determine the protein expression of Beclin 1and the ratio of LC3 Ⅱ to LC3Ⅰ,and real-time fluorescence quantitative PCR(qPCR) was tomeasure the mRNA expression of Atg6 and LC3B.Results:The cell viability,the protein level of Beclin 1,the ratio of LC3 Ⅱ/LC3 Ⅰ,and the mRNA expression levels of Atg 6 and LC3B in HGHL+ghrelin groupweresignificantly increased compared with HGHL group (P<0.05).MDC staining and immunofluorescence staining also showed increased deposition of autophagic vesicles and autophagic protein LC3B in HGHL+ghrelin group. Conclusion:Ghrelin could protect islet β cells from HGHL injury, which might be related to the enhancement of autophagy.

ObjectiveThis study aimed to investigate the effects of liraglutide on the body weight set point (BWSP) in diet‐induced obese rats and to determine the relationship between BWSP and hypothalamic arcuate nucleus (ARC) microglial activation.MethodsDiet‐induced obesity (DIO) rats were divided into three groups: continuous high‐fat diet (HFD) plus saline, HFD with liraglutide, and HFD with liraglutide pair feeding. Body weight, BWSP, inflammatory cytokines, suppressor of cytokine signaling 3, orexigenic/anorexigenic proteins, apoptosis, and microglia in the ARC were assessed. The effect of liraglutide on the Notch‐1 signaling pathway and its relationships with nuclear factor‐κB and p38 mitogen‐activated protein kinase were also investigated in a lipopolysaccharide (LPS)‐induced microglia activation model.ResultsLiraglutide reduced BWSP; reversed adverse changes in hypothalamic inflammation, suppressor of cytokine signaling 3, and apoptosis; and diminished microgliosis in DIO rats. The BWSP showed a linear correlation with ARC microglial density. Liraglutide inhibited LPS‐induced M1 microglial polarization and promoted microglial polarization to the M2 phenotype, diminishing inflammatory cytokine expression. Liraglutide inhibited Notch‐1 signaling pathway activation and decreased nuclear factor‐κB and p38 mitogen‐activated protein kinase pathway activation in LPS‐stimulated microglia.ConclusionsLiraglutide can reduce BWSP in DIO rats. There is a linear correlation between hypothalamic microgliosis and BWSP. Liraglutide reduces excessive microglial activation and inflammation, which may contribute to BWSP reduction.

Objective:To establish a simple and efficient method for the separation and purification of primary microglia.Methods:The cerebral cortex tissues were collected from one-day-old neonatal SD rats,and cells were obtained through trypsin digestion.The single cell suspension was then cultured.After the cells were confluent on the bottom of the culture flask,the culture flask was vigorously tapped for 3min,then centrifuged and the precipitate was collected and cultured again.The purity was identified by immunofluorescence. Results:The purity of microglia was 96.85％.Conclusion:Vigorously tapping the culture flask could achieve high yield and high purity of microglia,and it provided the foundation for the study of microglia in neurodegenerative diseases.

Primary pancreatic α, β, δ, and pancreatic polypeptide (PP) cells are reliable cell models for diabetes research. However, the separation and purification of these cells in living conditions remains an obstacle for researchers. The interaction of visible light with cellular molecules can produce Raman scattering, which can be analyzed to obtain cellular intrinsic molecular fingerprints. It has been speculated that primary pancreatic α, β, δ, and PP cells can be identified and separated from each other according to their spectral differences. To test this hypothesis, Raman spectra detection was performed on rat islet cells. Single islet cells identified by Raman scattering under living conditions were verified using immunohistochemistry. Thus, Raman data were acquired from a pure line of islet cells as a training sample and then used to establish the discriminant function. Then, using the principal component analysis–linear discriminate analysis (PCA-LDA) method, the four types of islet cells could be identified and discriminated by Raman spectroscopy. This study provides a label-free and noninvasive method for discriminating islet cell types in a randomly distributed mixed islet cell population via their physical properties rather than by using antibodies or fluorescence labeling.

Objective:To explore the differences of obese Sprague-Dawley (SD)rats model induced by lard oil high-fat (HF)diet or purified HF diet.Methods:SD weanling rats were randomly divided into three groups:D1 group,where rats were fed by lard oil HF diet;D2 group,where rats were fed by purified HF diet;C group,where rats were fed on chow.After 12 weeks,diet-induced obesity rats (top 33％ based on weight)were selected for further study,and the rest rats from group D1 and D2 were excluded.The food intake and weight were weighted daily and weekly,respectively.The subcutaneous,visceral and total fat contents of rats was measured by 256-row CT scan and the Lee index was calculated accordingly.The kidney,liver,testis,spleen and heart were weighted respectively.Serum leptin and insulin levels were quantified.The pathology in liver and adipose tissues were analyzed by HE staining.Oral glucose tolerance test (OGTT)was used to compare the glucose tolerance ability.Serum total cholesterol (T-CHO),high density lipoprotein (HDL-C),low density lipoprotein (LDL-C),triglyceride (TG)and inflammatory cytokines IL-6,TNF-α were detected as well.Results:After 12 weeks,the body weight,subcutaneous fat,visceral fat,total fat mass,wet weight of liver,kidney and heart,area under blood glucose curve and the levels of serum insulin,leptin,T-CHO,LDL-C,TG,IL-6 and TNF-α in group D2 were significantly increased compared to those of group C and group D1.HDL-C of group D2 was markedly lower than that in group C (P＜0.05).The visceral fat,total fat content and HDL-C in group D1 were significantly different from those of group C (P＜0.05).Steatosis and enlarged adipocyte were found in the livers of rats in group D1 and D2,and the lesions were more significant in group D2.Conclusion:Purified HF diet was more effective in inducing metabolic abnormalities,steatosis,peripheral chronic inflammation in obese SD rat models.But lard oil HF diet was more economical when only inducing visceral steatosis was required.

The examination of insulin (Ins) exocytosis at the single-cell level by conventional methods, such as electrophysiological approaches, total internal reflection imaging, and two-photon imaging technology, often requires an invasive microelectrode puncture or label. In this study, high concentrations of glucose and potassium chloride were used to stimulate β cell Ins exocytosis, while low concentrations of glucose and calcium channel blockers served as the blank and negative control, respectively. Laser tweezers Raman spectroscopy (LTRS) was used to capture the possible Raman scattering signal from a local zone outside of the cell edge. The results show that the frequencies of the strong signals from the local zones outside the cellular edge in the stimulated groups are greater than those of the control. The Raman spectra from the cellular edge, Ins and cell membrane were compared. Thus, local Ins exocytosis activity outside pancreatic β cells might be observed indirectly using LTRS, a non-invasive optical method.

Objective:This study was aimed to use hoarding behavior to measure the body weight set-point in rats and to discuss the effect of food restriction on the body weight set-point.Methods:Twelve male SD rats were randomly divided into a control group and a treatment group (n=6 per group).After measuring baseline set-point,the control group rats were given the same amount of food as baseline set point period and the treatment group rats were treated with food restriction (70％ of normal diet) for 6 weeks,and the rats were monitored for additional 6 weeks after ceasing the treatment.The amount of food hoarded by the rats and their body weight were measured 6 days per week.And the hoarding behavior was measured every other week at various body weights from 0 to 12 weeks.The body weight set-point of rats was calculated by linear regression equation.Results:The body weight set-point value in both groups were continuously increased from 0 to 12 weeks,but no significant difference was found between the two groups (P＞0.05).During food restriction period,body weight of the treatment group rats was significantly decreased (P＜0.01),however,their body weight gained after ceasing food restriction (P＜0.01).Conclusions:These results suggested that food restriction temporarily reduced body weight,rather than body weight set-point in rats.The behavioral method of the weight threshold to hoard food to assess body weight set point is accurate and reliable.

Objective To investigate the clinical significance of premenopausal patients with systemic lupus erythematosus (SLE) complicated with osteoporosis (OP) the RANKL changes. Methods Total of SLE and OP with prior diagnosis of premenopausal female patients the 26 cases of OP group, selected the premenopause patients with SLE without OP and 26 cases of N-OP group, another randomly selected the same period 26 cases of premenopausal female healthy subjects, and the level of RANKL was determined by ELISA method.Results The level of RANKL in SLE group was signiifcantly higher than that of the control group (P<0.05), patients with OP group RANKL levels were higher than N-OP group (P<0.05).Conclusion RANKL plays an important role in the occurrence of OP on SLE patients, so the measurement of the indicators, which can effectively for diagnosis for SLE with OP.

目的 探讨预防应用塞来昔布是否可以减少使用唑来磷酸治疗老年性骨质疏松症不良反应的发生.方法 选取2012年1月至2015年1月老年性骨质疏松症患者116例,年龄61 ～86岁.依据输注唑来磷酸前是否预防应用塞来昔布及布洛芬分为空白对照组、布洛芬组、塞来昔布组.观察输注唑来磷酸过程中及输液后3d内不良反应发生的情况.结果 ①塞来昔布组发热比例最少,塞来昔布组7例(14％)、布洛芬组8例(25％)、空白对照组13例(38.2％).②中热(38.1-39℃)、高热(＞39℃)的塞来昔布组比例显著低于其他两组,而低热(37.3-38℃)的塞来昔布组比例略高于其他两组.各组在不同发热程度的比例分别为:塞来昔布组:低热4例(8％)、中热2例(4％)、高热1例(2％);布洛芬组:低热3例(6.3％)、中热3例(9.4％)、高热3例(9.4％);空白对照组:低热2例(5.9％)、中热5例(14.7％)、高热6例(17.6％).三组发热程度比较有显著性差异(P＜0.05).③肌肉痛、骨骼关节疼痛、流感样症状、消化道症状、头晕、头痛、心悸等不良反应发生率均为塞来昔布组最低.除心悸、皮疹外,其他不良反应发生率在三组间均有显著性差异(P＜0.05).结论 预防性应用塞来昔布可以减少唑来磷酸不良反应的发生.

目的:探讨获得高质量、数量多的原代大鼠胰岛细胞的有效可靠的方法.方法:以V型胶原酶顺行灌注大鼠胆总管消化胰腺,以Histopaque 1077等密度区带离心法分离胰岛和胰腺腺泡组织.以双硫腙染色及免疫组化法分别鉴定提取物,胰岛素释放试验鉴定胰岛的活性.结果:经Histopaque 1077等密度区带离心法分离纯化,每只大鼠平均获取(251±26)枚胰岛.双硫腙染色后完整胰岛呈猩红色细胞团.胰岛单细胞培养及胰岛素释放试验起始2.8 mmol/L低糖培养胰岛素分泌量为(23.53±5.946) pmol/L,16.7 mmol/L高糖刺激后胰岛素分泌量为(32.61±4.085) pmol/L,最后恢复2.8 mmol/L低糖培养胰岛素分泌量为(27.81±3.616)pmol/L,3者之间比较差异均有统计学意义(P＜0.05),提示胰岛细胞活性良好.结论:用Histopaque 1077等密度区带离心法分离纯化原代大鼠胰岛细胞兼具有数量多、活性好、纯度高的优点,可继续用于研究.

目的:利用激光拉曼光谱系统无损区分贴壁生长的大鼠原代胰岛α、β细胞,并比较该方法与传统方法的优劣.方法:收集贴壁生长于定位格子培养皿中大鼠胰岛细胞的拉曼光谱、坐标及照片后应用免疫组化结合培养皿坐标定位确定胰岛细胞的分类,用主成分分析结合线性判别分析(PCA LDA)对两种胰岛细胞拉曼光谱进行分析,用线性判别分析法对两种细胞大小进行分析,两组间光谱及面积差异则采用两独立样本t检验.结果:贴壁生长的胰岛β细胞体积较α细胞大,差异具有统计学意义(P=0.001);然而单纯利用面积胰岛α细胞及胰岛β细胞正确区分率分别为84.5％、50.0％;胰岛α细胞与胰岛β细胞的拉曼光谱有共同特征峰同时也存在显著差异的波峰,即二者有不同的“细胞分子指纹”,且将该特点结合PCA-LDA的方法对胰岛α、β细胞进行区分正确率均为100％.结论:激光拉曼光谱系统可以成为快速无损区分胰岛α、β细胞的有效方法.

目的：探讨金花茶对亚急性衰老大鼠抗氧化和抗凋亡的作用。方法拟建立大鼠亚急性衰老模型后，予高、低浓度金花茶灌胃干预。测肝脏和睾丸组织超氧化物歧化酶（SOD），丙二醛（MDA）含量及 bax、bcl－2 mRNA 表达水平。结果与衰老模型组对比，高、低浓度金花茶组肝脏和睾丸组织 SOD 含量和 bcl－2 mRNA 表达显著升高（P＜0．05），且 MDA 含量和 bax mRNA 表达明显下降（P＜0．05）。高、低浓度金花茶组间比较，上述指标有统计学差异（P＜0．05）。结结论金花茶可以提高 SOD 含量和降低 MDA 含量，通过上调 bcl－2和下调 bax 表达水平，减慢肝脏和睾丸细胞的凋亡，对延缓机体衰老有重要作用。