Effects of Diabetes Education on Emotional Distress in Patients with Type 2 Diabetes—An Experimental Study

Objective: It was to evaluate the effect of diabetes education on emotional distress in type 2 diabetes patients treated with oral medications. Methods: The experimental study took place in Albania and overall, 200 type 2 diabetes patients were enrolled (in both groups, intervention, and control) treated with oral medications, having levels of Glycated hemoglobin HbA1c > 6.5% as well the absence of associated diseases such as dementia and psychiatric disorders. Patients were randomly selected from the medical registry of family physicians in the Tirana region. Patients were screened for the emotional distress before and after the intervention with the self-administered questionnaire Problem Areas in Diabetes PAID 5. In addition, the levels of HbA1c in % were evaluated before and after intervention in both groups. Only intervention group underwent four diabetes education sessions offered by trained nursing staff while the control group continued the previous regime. The questionnaire reliability analysis was estimated by the Cronbach alpha coefficient. To compare the groups the t-test was used and the value of p Results: Mean age of patients in intervention and control group was respectively 54.03 ±9.57 and 55.82 ± 7.86. Before and after health education PAID 5 scores for the intervention group were respectively 11.3 vs. 8.75 while for the control group 11.9 vs. 11.35, p = 0.018. Levels of HbA1c% before and after education for the intervention group were 7.02 vs. 6.2 while for the control group 6.9 vs. 6.8, p = 0.001. Positive and significant correlation (r = 0.321, p = 0.001) was between level of emotional distress and the age of the patients. Conclusions: The study found that besides better control of diabetes, additional education of diabetic patients seemed to significantly improve the level of emotional distress due to diabetes in diabetic patients.

Comparison of the effect of intravitreal bevacizumab and intravitreal fasudil on retinal VEGF,TNFα,and caspase 3 levels in an experimental diabetes model

AIM: To evaluate the influence of an intravitreal injection of bevacizumab and fasudil on the retinal vascular endothelial growth factor(VEGF),tumor necrosis factor alpha(TNFα),and caspase 3 levels in a diabetic rabbit model. · METHODS: The study included 6 healthy rabbits(Group 1),6 rabbits with experimentally induced diabetes mellitus(DM)(Group 2),7 rabbits with experimentally induced DM to which intravitreal bevacizumab was administered(Group 3),and 7 rabbits with experimentally induced DM to which intravitreal fasudil was administered(Group 4). An intravitreal injection of 1.25mg/50μL bevacizumab in the right eye of rabbits in Group 3 and an intravitreal injection of 0.0064mg/50μL fasudil in the right eye of rabbits in Group 4 were administered on day 21 after the induction of DM. The studied eyes of the rabbits were enucleated three days after the intravitreal injection. The TNFα,VEGF,and caspase 3 levels were determined using the ELISA method. · RESULTS: There was a statistically significant difference in the VEGF and caspase 3 levels between groups(P =0.005 and P =0.013,respectively),but the TNF α level did not differ significantly between groups(P = 0.792). It was found that VEGF levels were significantly lower in Group 1 and Group 3 than in Group 2 using the Mann-Whitney U test with the Bonferroni correction(P = 0.004 for both comparison). There was no statistically significant difference between other groups with regard to VEGF levels(the P value ranged between 0.015 and 0.886). Although the P values of the caspase 3 levels were 0.015 for Group 1 and Group 4,0.038 for Group 2and Group 3,and 0.018 for Group 3 and Group 4,these P values remained above the threshold P value of 0.0083,which was the statistically significant level for post hoc tests. ·CONCLUSION: An intravitreal injection of bevacizumab decreased both the VEGF level,which plays a role in angiogenesis,and the caspase 3 level,which plays a role in apoptosis. Although not as effective as bevacizumab,fasudil had a beneficial effect on the VEGF levels but significantly increased the caspase 3 levels.

Potential application of Nardostachyos Radix et Rhizoma-Rhubarb for the treatment of diabetic kidney disease based on network pharmacology and cell culture experimental verification

BACKGROUND Diabetic kidney disease(DKD)is one of the serious complications of diabetes mellitus,and the existing treatments cannot meet the needs of today's patients.Traditional Chinese medicine has been validated for its efficacy in DKD after many years of clinical application.However,the specific mechanism by which it works is still unclear.Elucidating the molecular mechanism of the Nardostachyos Radix et Rhizoma-rhubarb drug pair(NRDP)for the treatment of DKD will provide a new way of thinking for the research and development of new drugs.AIM To investigate the mechanism of the NRDP in DKD by network pharmacology combined with molecular docking,and then verify the initial findings by in vitro experiments.METHODS The Traditional Chinese Medicine Systems Pharmacology(TCMSP)database was used to screen active ingredient targets of NRDP.Targets for DKD were obtained based on the Genecards,OMIM,and TTD databases.The VENNY 2.1 database was used to obtain DKD and NRDP intersection targets and their Venn diagram,and Cytoscape 3.9.0 was used to build a"drug-component-target-disease"network.The String database was used to construct protein interaction networks.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and Gene Ontology analysis were performed based on the DAVID database.After selecting the targets and the active ingredients,Autodock software was used to perform molecular docking.In experimental validation using renal tubular epithelial cells(TCMK-1),we used the Cell Counting Kit-8 assay to detect the effect of NRDP on cell viability,with glucose solution used to mimic a hyperglycemic environment.Flow cytometry was used to detect the cell cycle progression and apoptosis.Western blot was used to detect the protein expression of STAT3,p-STAT3,BAX,BCL-2,Caspase9,and Caspase3.RESULTS A total of 10 active ingredients and 85 targets with 111 disease-related signaling pathways were obtained for NRDP.Enrichment analysis of KEGG pathways was performed to determine advanced glycation end products(AGEs)-receptor for AGEs(RAGE)signaling as the core pathway.Molecular docking showed good binding between each active ingredient and its core targets.In vitro experiments showed that NRDP inhibited the viability of TCMK-1 cells,blocked cell cycle progression in the G0/G1 phase,and reduced apoptosis in a concentrationdependent manner.Based on the results of Western blot analysis,NRDP differentially downregulated p-STAT3,BAX,Caspase3,and Caspase9 protein levels(P<0.01 or P<0.05).In addition,BAX/BCL-2 and p-STAT3/STAT3 ratios were reduced,while BCL-2 and STAT3 protein expression was upregulated(P<0.01).CONCLUSION NRDP may upregulate BCL-2 and STAT3 protein expression,and downregulate BAX,Caspase3,and Caspase9 protein expression,thus activating the AGE-RAGE signaling pathway,inhibiting the vitality of TCMK-1 cells,reducing their apoptosis.and arresting them in the G0/G1 phase to protect them from damage by high glucose.

Insulin is necessary for the hypertrophic effect of cholecystokinin-octapeptide following acute necrotizing experimental pancreatitis

AIM:In previous experiments we have demonstrated that by administering low doses of cholecystokinin-octapeptide (CCK-8),the process of regeneration following L-arginine (Arg)-induced pancreatitis is accelerated.In rats that were also diabetic(induced by streptozotocin,STZ),pancreatic regeneration was not observed.The aim of this study was to deduce whether the administration of exogenous insulin could in fact restore the hypertrophic effect of CCK-8 in diabetic-pancreatitic rats. METHODS:Male Wistar rats were used for the experiments. Diabetes mellitus was induced by administering 60mg/kg body mass of STZ intraperitoneally(i.p.),then,on d 8, pancreatitis was induced by 200mg/100 g body mass Arg i.p.twice at an interval of 1 h.The animals were injected subcutaneously twice daily(at 7 a.m.and 7 p.m.)with 1 μg/kg of CCK-8 and/or 2 IU mixed insulin(300g/L short- action and 700g/L intermediate-action insulin) for 14 d after pancreatitis induction.Following this the animals were killed and the serum amylase,glucose and insulin levels as well as the plasma glucagon levels,the pancreatic mass/body mass ratio(pm/bm),the pancreatic contents of DNA,protein,amylase,lipase and trypsinogen were measured.Pancreatic tissue samples were examined by light microscopy on paraffin-embedded sections. RESULTS:In the diabetic-pancreatitic rats treatment with insulin and CCK-8 significantly elevated pw/bm and the pancreatic contents of protein,amylase and lipase vs the rats receiving only CCK-8 treatment.CCK-8 administered in combination with insulin also elevated the number of acinar cells with mitotic activities,whereas CCK-8 alone had no effect on laboratory parameters or the mitotic activities in diabetic-pancreatitic rats. CONCLUSION:Despite the hypertrophic effect of CCK-8 being absent following acute pancreatitis in diabetic-rats, the simultaneous administration of exogenous insulin restored this effect.Our results clearly demonstrate that insulin is necessary for the hypertrophic effect of low-doses of CCK-8 following acute pancreatitis.

Changes of activity of liver glycogen synthase in experimental diabetes mellitus rats

OBJECTIVE: To study the activity of liver glycogen synthase in analogous model of NIDDM rats. METHODS: Male Wistar rats were injected with low dose of streptozotocin (STZ) (30 mg/kg body weight) via tail vena and those animals with glucose tolerance impaired and level of insulin equal to or higher than that of the controls at 18th week were taken as the analogous rat model of NIDDM. The activity of liver glycogen synthase (GS) was assayed at the end of experiment. RESULTS: Type I-enzyme: 0.18 +/- 0.06 mumol/min.g versus 0.24 +/- 0.09 mumol/min.g, P

Biomechanics of Bone in Experimental Diabetic rats.

The biomechanics of bone In diabetics and its response to insulin treatment was not clear.We investigated the impace of alloxan-induced diabetes and insulin treatment on the biomechanical characteristics of bones．Sprague-Dawley rats were divided into three groups:

Effects of glucagon-like peptide 1 analogs in combination with insulin on myocardial infarct size in rats with type 2 diabetes mellitus

AIM To evaluate the effects of glucagon-like peptide-1 analogs(GLP-1 a) combined with insulin on myocardial ischemiareperfusion injury in diabetic rats.METHODS Type 2 diabetes mellitus(T2 DM) was induced in maleWistar rats with streptozotocin(65 mg/kg) and verified using an oral glucose tolerance test. After anesthesia, the left coronary artery was occluded for 40 min followed by 80 min reperfusion. Blood glucose level was measured during surgery. Rats were randomized into six groups as follows:(1) control rats;(2) insulin(0.1 U/kg) treated rats prior to ischemia;(3) insulin(0.1 U/kg) treated rats at reperfusion;(4) GLP-1 a(140 mg/kg) treated rats prior to ischemia;(5) GLP-1 a(140 mg/kg) treated rats at reperfusion; and(6) rats treated with GLP-1 a(140 mg/kg) prior to ischemia plus insulin(0.1 U/kg) at reperfusion. Myocardial area at risk and infarct size was measured planimetrically using Evans blue and triphenyltetrazolium chloride staining, respectively.RESULTS There was no significant difference in the myocardial area at risk among groups. Insulin treatment before ischemia resulted in a significant increase in infarct size(34.7% ± 3.4% vs 18.6% ± 3.1% in the control rats, P < 0.05). Post-ischemic administration of insulin or GLP-1 a had no effect on infarct size. However, pre-ischemic administration of GLP-1 a reduced infarct size to 12% ± 2.2%(P < 0.05). The maximal infarct size reduction was observed in the group treated with GLP-1 a prior to ischemia and insulin at reperfusion(8% ± 1.6%, P < 0.05 vs the control and GLP-1 a alone treated groups).CONCLUSION GLP-1 a pre-administration results in myocardial infarct size reduction in rats with T2 DM. These effects are maximal in rats treated with GLP-1 a pre-ischemia plus insulin at reperfusion.

Experimental study of tibial transverse bone transfer on wound healing of diabetic rabbit leg ulcer

Objective To investigate the efficacy and mechanism of tibial transverse bone transfer in the treatment of lower limb ulcer wounds in diabetic rabbits.Methods Animal models of diabetic foot lower extremity ischemia were made,and grouped Animal models of diabetic foot lower extremity ischemia were made,and grouped control studies were designed to evaluate the time of wound healing and the expression level of vascular endothelial growth factor(VEGF),platelet derived growth factor(PDGF),and epidermal cell growth factor(EGF)in the wound during the same period of time.Results In animal experiments,transverse tibial bone transfer can shorten the healing time of diabetic ulcer and increase the expression of wound growth factor(VEGF,PDGF,EGF).Conclusion In animal experiments,diabetes can inhibit the expression of growth factors(VEGF,PDGF,EGF)in ulcer wounds,and tibial transverse bone transfer can promote wound healing.The mechanism may be related to the increased expression of VEGF,PDGF and EGF in the wound surface.

The impact of Hypoglycemic Ziyabiti Tablets (HZT) on diabetes mellitus rats and its activity in cultured rat liver cells

Objective Hypoglycemic Ziyabiti Tablets(HZT)is a traditional multicomponent treatment for diabetes in Xinjiang Uyghur Traditional Medicine.The present study aimed to investigate the effect of HZT in diabetic rats and its activity in cultured liver cells to investigate the relative mechanisms.Methods 10 days high-fat diet fed rats were intraperitoneally injected with alloxan(ALX)at next two subsequent days to induce diabetes mellitus(DM).Then were divided into 5 groups:saline,positive DM control and DM groups treated with different doses of HZT.Fasting blood glucose(FBG),total cholesterol(TC),total triglycerides(TG),high-density lipoprotein(HDL-C),fasting insulin(FI),insulin secretion(IS)and insulin sensitivity index(ISI)were measured.The IC_(50) of HZT in L-02 cells was determined by MTT assay,in intact and in paracetamol-induced liver injury(Par),on lactate dehydrogenase(LDH)activity and on glucose consumption.Results HZT decreased FBG and TC(P <0.05),increased IS(P <0.05)and at 440 mg·kg^(-1)·d^(-1) increased FI(P < 0.01).In vitro,HZT at 0.1,0.2,and 0.4 mg/mL decreased LDH activity and promoted glucose consumption.Conclusion The hypoglycemic mechanism of HZT is possibly related to increased insulin secretion from the pancreas and increased utilization of glucose by the liver.

PKC-α介导的线粒体功能障碍在妊娠糖尿病致胚胎神经管缺陷中的机制研究

目的:探讨蛋白激酶C-α(PKC-α)介导的线粒体功能障碍在妊娠糖尿病致胚胎神经管缺陷(NTD)中的作用。方法:将雌性C57BL/6J小鼠随机分为对照组、NTD组、NTD+sh-PKC-α组,每组6只。除对照组外,NTD组、NTD+sh-PKC-α组建立糖尿病胚胎病小鼠模型。NTD+sh-PKC-α组分别在胚胎第1天(E1)、第4天(E4)和第8天(E8)时,经尾静脉向小鼠注射100μL携带sh-PKC-α的慢病毒(1×10^(8) TU/mL)。在第11.5天(E11.5)时,从子宫中解剖出胚胎进行分析。采用透射电子显微镜观察神经管中线粒体。采用蛋白免疫印迹法(Western Blot)分析胚胎神经管中PKC-α、抗微管相关蛋白1轻链3(LC3)和抗溶酶体相关膜蛋白2(LAMP2)蛋白表达。体外构建PKC-α敲低C17.2小鼠神经干细胞,并在低(5 mmol/L)或高(25 mmol/L)葡萄糖条件下培养,采用免疫荧光染色检测细胞自噬体变化情况。结果:在NTD组中,胚胎神经管中线粒体肿胀,嵴消失。与对照组相比,NTD组胚胎神经管中LC3Ⅱ和LAMP2蛋白表达降低(P<0.05),PKC-α蛋白表达上调(P<0.01)。通过沉默小鼠胚胎神经管中PKC-α表达,胚胎中LC3Ⅱ和LAMP2蛋白表达增加(P<0.05)。此外,NTD+sh-PKC-α组小鼠胚胎中的NTD发生率低于NTD组(P<0.01)。在高糖条件下,C17.2神经干细胞中LC3Ⅱ、LAMP2蛋白表达水平和Cyto-ID染色点的数量均低于正常葡萄糖条件下水平(P<0.05)。PKC-α蛋白敲低增加了高糖条件下的LC3Ⅱ、LAMP2蛋白表达水平和Cyto-ID染色点的数量(P<0.05)。结论:PKC-α介导的线粒体功能障碍与母体糖尿病诱导的NTD胚胎相关,通过沉默母体PKC-α减弱了神经管自噬抑制导致的NTD形成。

基于网络药理学及细胞实验验证探讨补肾益心片治疗糖尿病肾病的作用机制

【目的】探讨补肾益心片治疗糖尿病肾病的作用机制。【方法】应用网络药理学预测补肾益心片治疗糖尿病肾病的关键靶点。构建高糖处理MPC-5细胞模型,应用酶联免疫吸附法(ELISA)、实时定量聚合酶链反应(RT-qPCR)法和Western Blot法验证补肾益心片治疗糖尿病肾病的潜在作用靶点及通路。【结果】获得补肾益心片24个有效成分。补肾益心片治疗糖尿病肾病涉及靶点131个,通路富集分析结果显示关键靶点可能主要集中在炎症通路、脂质与动脉粥样硬化等。进一步实验验证结果发现:与高糖组比较,补肾益心片含药血清中、高剂量组MPC-5细胞上清中基质金属蛋白酶1(MMP-1)分泌水平升高(P<0.001),组织金属蛋白酶抑制因子1(TIMP-1)和转化生长因子β1(TGF-β1)分泌水平降低(P<0.001);补肾益心片含药血清高剂量组MMP-1和BCL-2蛋白及mRNA表达水平升高(P<0.001),磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(AKT)蛋白及mRNA表达水平降低(P<0.001)。【结论】补肾益心片可能是通过调节PI3K/AKT/BCL-2通路抑制肾小球系膜细胞增殖,促进细胞外基质的降解,从而发挥对糖尿病肾病的治疗作用。

海参皂苷调节SIRT1/NF-κB信号通路对糖尿病大鼠血管炎症的影响

目的:探讨海参皂苷(SSC)对糖尿病大鼠血管炎症的影响及潜在机制。方法:SD大鼠采用高糖高脂饮食和链脲佐菌素(STZ)腹腔注射构建糖尿病模型。实验分为对照(Control)组、模型(model)组、二甲双胍组(600 mg/kg)、SSC组(30 mg/kg)和SSC+沉默信息调节因子1(SIRT1)抑制剂EX-527组(SSC 30 mg/kg+EX-5275 mg/kg)。给予相应的药物干预4周后,检测各组大鼠空腹血糖(FBG)和空腹胰岛素(FINS)水平,计算胰岛素抵抗指数(HOMA-IR);酶联免疫吸附法(ELISA)检测血清血管细胞黏附分子-1(VCAM-1)、单核细胞趋化因子-1(MCP-1)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)水平;苏木精-伊红(HE)染色观察主动脉病理学变化;蛋白免疫印迹法(Western Blot)检测主动脉SIRT1/核因子-κB(NF-κB)通路相关蛋白的表达。结果:与Control组比较,model组大鼠FBG、FINS、HOMA-IR、血清VCAM-1、MCP-1、TNF-α、IL-1β水平、胞核NF-κB p65蛋白水平升高(P<0.05),体质量、主动脉SIRT1、核因子κB抑制蛋白α(IκBα)蛋白水平降低(P<0.05);与model组比较,SSC组和二甲双胍组大鼠FBG、FINS、HOMA-IR、血清VCAM-1、MCP-1、TNF-α、IL-1β水平、胞核NF-κB p65的蛋白水平降低(P<0.05),体质量、主动脉SIRT1、IκBα蛋白水平升高(P<0.05);与SSC组比较,SSC+EX-527组大鼠FBG、FINS、HOMA-IR、血清VCAM-1、MCP-1、TNF-α、IL-1β水平、胞核NF-κB p65蛋白水平升高(P<0.05),体质量、主动脉SIRT1、IκBα蛋白水平降低(P<0.05)。结论:SSC可减轻糖尿病大鼠血管炎症,其作用机制可能与上调SIRT1表达,抑制NF-κB的活化有关。

基于PI3K/AKT/mTOR信号通路研究破淤汤对缺血性糖尿病足溃疡大鼠促血管新生的作用

目的:基于PI3K/AKT/mTOR信号通路探索破淤汤对缺血性糖尿病足溃疡大鼠促血管新生的作用机制。方法:将造模成功的缺血性糖尿病足溃疡大鼠随机分为非药物组、西洛他唑组、破淤汤低剂量组、破淤汤中剂量组、破淤汤高剂量组,各组给予相应药物或非药物干预后,通过创面肉芽组织HE染色观察组织结构,通过CD31免疫组化染色观察血管新生,通过酶联免疫吸附(ELISA)双抗体夹心法检测血清中VEGFR-2水平,通过Western免疫印迹法检测PI3K/Akt/mTOR信号通路关键蛋白表达情况。结果:HE染色结果显示低剂量破淤汤组与非药物组比较,新生毛细血管明显增多,同样含大量炎细胞;中、高剂量破淤汤组较非药物组改善明显,新生毛细血管多,炎症细胞少,接近于正常对照组。与非药物组比较,西洛他唑组及低、中、高剂量破淤汤组大鼠糖尿病足溃疡肉芽组织微血管数目、P-PI3K、P-AKT、P-mTOR表达水平及大鼠血清VEGFR2表达量均升高(P均﹤0.05)。结论:破淤汤可能通过促进缺血溃疡组织VEGER-2蛋白表达,促进PI3K/AKT/mTOR信号通路关键蛋白的表达来促进缺血溃疡组织的血管新生。

糖络宁减轻大鼠背根神经节细胞炎症反应的作用机制研究

目的:以施万细胞外泌体为切入点,观察糖络宁含药血清对体外高糖诱导的大鼠背根神经节细胞炎症反应的影响。方法:采用糖络宁含药血清干预高糖培养的施万细胞,分为正常组、高糖组、糖络宁组,干预48 h后,提取各组细胞上清液中外泌体,将各组外泌体分别干预正常糖和高糖条件培养的大鼠背根神经节细胞,分为无外泌体-正常糖组,正常外泌体-正常糖组、高糖外泌体-正常糖组、糖络宁外泌体-正常糖组、无外泌体-高糖组、正常外泌体高糖组、高糖外泌体-高糖组、糖络宁外泌体-高糖组8个组,48 h后,收集各组细胞并进行相关检测。CCK-8法检测各组细胞活力,酶联免疫法检测各组细胞上清液中炎症介质(TNF-α、IL-6、IL-1β)的蛋白表达情况,实时荧光定量PCR检测各组细胞炎症介质的表达情况。结果:与正常外泌体-正常糖组比较,高糖外泌体-正常糖组细胞活力下降(P<0.05),炎症介质的基因和蛋白表达均增加(P<0.05);而与高糖外泌体-正常糖组比较,糖络宁外泌体-正常糖组细胞活力有所升高(P<0.05),各炎症介质的基因和蛋白表达均有所下调(P<0.05)。高糖条件培养的背根神经节细胞中,不同组外泌体干预表现出类似的作用效果;结论:糖络宁能通过调控施万细胞外泌体减轻大鼠背根神经节细胞炎症反应。

益消方经转化生长因子β1信号通路干预糖尿病合并脂肪肝的实验研究

目的:探索益消方对糖尿病合并脂肪肝的影响和作用机制。方法:观察组为12周龄雄性自发糖尿病(KKAy)小鼠,按照随机数字表法随机分为模型组、中药组(益消方干预)、西药组(氯沙坦干预);对照组为同周龄雄性近交系(C57BL/6J)小鼠。干预周期为12周。观察干预前后血糖、血脂-、肝功能变化,肝脏组织进行病理染色,检测肝脏组织转化生长因子β_(1)(TGF-β_(1))信号通路蛋白和信使核糖核酸(mRNA)的变化。结果:与模型组比较,益消方干预8周体质量、肝重指数显著降低(P<0.05,P<0.01),干预第4、8、12周空腹血糖显著降低(P<0.01),干预8周胆固醇、三酰甘油、低密度脂蛋白胆固醇、谷丙转氨酶水平降低(P<0.05,P<0.01)。病理形态方面益消方干预4周脂肪变性评分下降(P<0.05)。益消方干预12周后,TGF-β_(1)及其信号蛋白、mRNA的表达均显著下调(P<0.05,P<0.01)。结论:益消方可显著改善KKAy小鼠的肥胖和糖脂代谢紊乱,调节TGF-β_(1)信号通路蛋白和基因表达,减轻肝脏脂肪变性。

基于网络药理学和试验验证探讨茵陈蒿汤治疗糖尿病的作用机制

【目的】通过网络药理学、分子对接和试验验证的方法探讨茵陈蒿汤(YCHD)治疗糖尿病(DM)的作用机制。【方法】利用TCMSP、ETCM、UniProt、SwissTargetPrediction数据库及文献查阅等查找YCHD的活性成分和相关作用靶点;利用OMIM、PharmGkb、TTD、DrugBank数据库获取DM的疾病靶点。构建YCHD和DM的蛋白互作(PPI)网络图,对关键作用靶点进行GO功能和KEGG通路富集分析。采用AutoDockTool 1.5.7软件进行分子对接,预测活性成分和核心靶点的结合能。将细胞分为对照组(Control)、模型组(Model)及YCHD含药血清低(YCHD-L)、中(YCHD-M)、高(YCHD-H)剂量组,通过体外细胞试验检测诱导型一氧化氮合酶(iNOS)、环氧化酶2(COX2)、Toll样受体4(TLR4)、核因子κB(NF-κB)p65、白细胞介素-1β(IL1β)、IL6的mRNA表达水平,以及iNOS、COX2、髓样分化因子88(MyD88)、NF-κB p65蛋白表达水平,并对关键通路进行验证。【结果】共获得YCHD活性成分147个,DM疾病靶点486个,YCHD作用于DM的相关靶点基因57个,其中胰岛素(INS)、IL1β、肿瘤坏死因子(TNF)、B细胞中κ轻型多肽基因增强子的核因子1(NFKB1)、信号传导子和转录活化子1(STAT1)、白蛋白(ALB)、TLR4、IL1A、IL10、IL8等为关键靶基因。根据Counts值筛选出GO前十的条目中包含生物过程1条、细胞组分7条、分子功能2条,主要包括RNA聚合酶Ⅱ启动子转录的正调控、内质网膜、质膜、胞外区、核质、胞质溶胶、细胞质、膜的组成部分、相同蛋白质结合和蛋白质结合。KEGG通路富集分析显示,与NOD样受体、NF-κB、Toll样受体等炎症信号通路密切相关。分子对接结果显示,YCHD的活性有效成分与主要靶点具有良好的结合能力。体外试验结果表明,与对照组相比,模型组细胞TLR4、iNOS、COX2、NF-κB p65、IL1β和IL6 mRNA表达水平及iNOS、COX2、MyD88、NF-κB p65蛋白表达水平均显著升高(P<0.05);与模型组相比,YCHD-L、YCHD-M、YCHD-H组细胞iNOS、COX2、IL1β和IL6 mRNA表达水平及iNOS、COX2蛋白表达水平均显著降低(P<0.05),YCHD-M、YCHD-H组细胞TLR4、NF-κB p65 mRNA水平显著降低(P<0.05),YCHD-H组细胞MyD88、NF-κB p65蛋白表达水平显著降低(P<0.05)。【结论】YCHD可能通过多成分、多靶点,调控TLR4、NF-κB等多条炎症信号通路,发挥抗DM的作用。以YCHD-H组对炎症信号通路相关指标的抑制最明显。

基于网络药理学和实验验证探讨玉泉胶囊治疗糖尿病肾病的作用机制

[目的]运用网络药理学和体外实验探讨玉泉胶囊治疗糖尿病肾病的潜在作用机制。[方法]借助中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)和中医药综合数据库(Traditional Chinese Medicine Integrated Database,TCMID)筛选玉泉胶囊有效成分及靶点基因,基因组注释数据平台(Genome Annotation Database Platform,GeneCards)、疾病基因网络(Disease Gene Network,DisGeNET)数据库和治疗靶点数据库(Therapeutic Target Database,TTD)筛选糖尿病肾病的疾病相关基因,STRING 11.5数据库构建蛋白互作(protein-protein interaction,PPI)网络,并利用Cytoscape 3.7.2软件可视化,利用Cytoscape 3.7.2软件的CytoHubba和MCODE插件筛选关键化合物与核心靶点,利用R软件(ClusterProfiler包)进行基因本体(gene ontology,GO)富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路富集分析,利用AutoDock和PyMOL软件进行分子对接验证,最后对网络药理学预测结果进行体外实验验证。[结果]玉泉胶囊抗糖尿病肾病活性成分有177个,活性成分作用靶点共477个,作用于糖尿病肾病的靶点有135个,核心靶点包括丝氨酸/苏氨酸激酶1(serine/threonine kinase 1,AKT1)、白细胞介素-1β(interleukin-1 beta,IL1B)、过氧化氢酶(catalase,CAT)、一氧化氮合酶(nitric oxide synthase 3,NOS3)、瘦素(leptin,LEP)、胰岛素样生长因子1(insulin-like growth factor 1,IGF1)和趋化因子配体2(C-C motif chemokine ligand 2,CCL2);GO富集分析共得到相关条目2557个,其中生物过程1415个、细胞组成657个、分子功能48个;KEGG通路分析结果显示玉泉胶囊作用于糖尿病肾病的靶点富集在66条通路上,其中环磷酸腺苷(cyclic adenosine monophosphate,cAMP)信号通路、高级糖基化终末产物-受体(advanced glycation end products-receptor for advanced glycation end products,AGE-RAGE)信号通路在糖尿病并发症中的作用等可能是玉泉胶囊治疗糖尿病肾病的关键通路。分子对接结果显示核心靶点与关键化合物均能较好地结合。实验验证显示,CCL2蛋白水平呈药物剂量依赖性下降,CAT、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOs)蛋白水平呈药物剂量依赖性上升。[结论]玉泉胶囊治疗糖尿病肾病具有多成分、多靶点的特点,其机制可能与下调CCL2、上调CAT、eNOs的蛋白表达,抑制糖尿病肾病的氧化应激,调节关键通路相关。

综合设计性实验在糖尿病护理实验教学中的应用

目的 探讨综合设计性实验在高职护理学生糖尿病护理实验教学中的应用效果。方法 采用整群抽样法,以广西某高校2020级及2021级护理高职学生作为研究对象。选取2020级护理高职学生225人作为对照组,2021级护理高职学生176人作为实验组。在糖尿病护理实验课教学中,对照组采用常规教学方法,实验组采用综合设计性实验。利用量表比较两组学生自主学习能力得分、合作问题解决能力得分及批判性思维能力得分。结果 实验组护理高职学生自主学习能力测评量表评分、合作问题解决能力评分、批判性思维能力7个维度评分与总分均较对照组高(P<0.05)。结论 综合设计性实验教学有助于护理高职学生自主学习能力、合作问题解决能力及批判性思维能力的培养。

miR-200a通过上调Nrf2对老年糖尿病心肌病大鼠心肌的保护作用

目的:探讨微小RNA-200a(miR-200a)对老年糖尿病大鼠心肌的保护作用及机制。方法:采用高脂饮食+链脲菌素(STZ)诱导大鼠糖尿病心肌病模型,分为对照组和STZ组。通过注射携带miR-200a腺相关病毒9(AAV9)在心肌内过表达miR-200a,分为对照+AAV9-control组、对照+AAV9-miR-200a组、STZ+AAV9-control组、STZ+AAV9-miR-200a组,每组8只。体外培养大鼠心肌细胞H9c2,分为对照组(5.5 mmol/L葡萄糖培养24 h)、高糖+AAV9-control+si RNA组(转染AAV9-control和si RNA 48 h后用33 mmol/L葡萄糖培养24 h)、高糖+AAV9-control+si核因子E2相关因子2(Nrf2)组(转染AAV9-control和si Nrf248 h后使用33 mmol/L葡萄糖培养24 h)、高糖+AAV9-miR-200a+siRNA组(转染AAV9-miR-200a和si RNA 48 h后用33 mmol/L葡萄糖培养24 h)、高糖+AAV9-miR-200a+si Nrf 2组(转染AAV9-miR-200a和si Nrf248 h后用33 mmol/L葡萄糖培养24 h)。使用酶联免疫吸附法(ELISA)试剂盒检测超氧化物歧化酶(SOD)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、丙二醛(MDA)含量;实时荧光定量聚合酶链式反应(PCR)检测miR-200a表达水平,蛋白免疫印迹法检测相关蛋白表达水平;2′,7′-二氯二氢荧光素二乙酸酯(DCFH-DA)染色检测细胞活性氧(ROS)水平;免疫荧光检测Nrf2表达。结果:与对照组比较,STZ组心肌组织miR-200a表达下调(P<0.05)。与对照+AAV9-control组比较,STZ+AAV9-control组心肌组织LDH、CK-MB、MDA水平升高,SOD水平、血红素氧化酶1(HO-1)、细胞核Nrf2蛋白、细胞质Nrf2蛋白表达及Nrf2荧光密度降低(P<0.05)。与STZ+AAV9-control组比较,STZ+AAV9-miR-200a组心肌组织LDH、CK-MB、MDA水平及TUNEL阳性细胞率降低,SOD水平、HO-1、细胞核Nrf2蛋白、细胞质Nrf2蛋白表达及Nrf2荧光密度升高(P<0.05)。高糖处理可降低H9c2细胞存活率,提高ROS、MDA、LDH水平(P<0.05);在此基础上转染miR-200a后上述指标被抑制;同时转染miR-200a和si Nrf2后,ROS、MDA、LDH升高,细胞存活率降低(P<0.05)。结论:miR-200a提高Nrf2表达从而减轻高糖诱导的心肌细胞氧化应激和凋亡,这一效应与Nrf2核易位的促进及下游抗氧化应激信号有关。