Mechanisms of Dangua Fang(丹瓜方)in multi-target and multi-method regulation of glycolipid metabolism based on phosphoproteomics

OBJECTIVE:To explore the mechanism of Dangua Fang(丹瓜方,DGR)in multi-target and multi-method regulation of glycolipid metabolism based on phosphoproteomics.METHODS:Sprague-Dawley rats with normal glucose levels were randomly divided into three groups,including a conventional diet control group(Group A),high-fat-highsugar diet model group(Group B),and DGR group(Group C,high-fat-high-sugar diet containing 20.5 g DGR).After 10 weeks of intervention,the fasting blood glucose(FBG),2 h blood glucose[PBG;using the oral glucose tolerance test(OGTT)],hemoglobin A1c(HbA1c),plasma total cholesterol(TC),and triglycerides(TG)were tested,and the livers of rats were removed to calculate the liver index.Then,hepatic portal TG were tested using the Gross permanent optimization-participatiory action planning enzymatic method and phosphoproteomics was performed using liquid chromatography with tandem mass spectrometry(LC-MS/MS)analysis followed by database search and bioinformatics analysis.Finally,cell experiments were used to verify the results of phosphoproteomics.Phosphorylated mitogen-activated protein kinase kinase kinase kinase 4(MAP4k4)and phosphorylated adducin 1(ADD1)were detected using western blotting.RESULTS:DGR effectively reduced PBG,TG,and the liver index(P<0.05),and significantly decreased HbA1c,TC,and hepatic portal TG(P<0.01),showed significant hematoxylin and eosin(HE)staining,red oil O staining,and Masson staining of liver tissue.The total spectrum was 805334,matched spectrum was 260471,accounting for accounting 32.3%,peptides were 19995,modified peptides were 14671,identified proteins were 4601,quantifiable proteins were 4417,identified sites were 15749,and quantified sites were 14659.Based on the threshold of expression fold change(>1.2),DGR upregulated the modification of 228 phosphorylation sites involving 204 corresponding function proteins,and downregulated the modification of 358 phosphorylation sites involving 358 corresponding function proteins,which included correcting 75 phosphorylation sites involving 64 corresponding function proteins relating to glycolipid metabolism.Therefore,DGR improved biological tissue processes,including information storage and processing,cellular processes and signaling,and metabolism.The metabolic functions regulated by DGR mainly include energy production and conversion,carbohydrate transport and metabolism,lipid transport and metabolism,inorganic ion transport and metabolism,secondary metabolite biosynthesis,transport,and catabolism.In vitro phosphorylation validation based on cell experiments showed that the change trends in the phosphorylation level of MAP4k4 and ADD1 were consistent with that of previous phosphoproteomics studies.CONCLUSION:DGR extensively corrects the modification of phosphorylation sites to improve corresponding glycolipid metabolism-related protein expression in rats with glycolipid metabolism disorders,thereby regulating glycolipid metabolism through a multi-target and multi-method process.

Mechanisms of Dangua Recipe in Improving Glycolipid Metabolic Disorders Based on Transcriptomics

Objective:To explore the mechanisms of Dangua Recipe(DGR)in improving glycolipid metabolism based on transcriptomics.Methods:Sprague-Dawley rats with normal glucose level were divided into 3 groups according to a random number table,including a conventional diet group(Group A),a DGR group(Group B,high-calorie diet+20.5 g DGR),and a high-calorie fodder model group(Group C).After 12 weeks of intervention,the liver tissue of rats was taken.Gene sequence and transcriptional analysis were performed to identify the key genes related to glycolipid metabolism reflecting DGR efficacy,and then gene or protein validation of liver tissue were performed.Nicotinamide phosphoribosyl transferase(Nampt)and phosphoenolpyruvate carboxykinase(PEPCK)proteins in liver tissues were detected by enzyme linked immunosorbent assay,fatty acid synthase(FASN)protein was detected by Western blot,and fatty acid binding protein 5(FABP5)-mRNA was detected by quantitative real-time polymerase chain reaction.Furthermore,the functional verification was performed on the diabetic model rats by Nampt blocker(GEN-617)injected in vivo.Hemoglobin A1c(HbA1c),plasma total cholesterol and triglycerides were detected.Results:Totally,257 differentialdominant genes of Group A vs.Group C and 392 differential-dominant genes of Group B vs.Group C were found.Moreover,11 Gene Ontology molecular function terms and 7 Kyoto Encyclopedia of Genes and Genomes enrichment pathways owned by both Group A vs.Group C and Group C vs.Group B were confirmed.The liver tissue target validation showed that Nampt,FASN,PEPCK protein and FABP5-mRNA had the same changes consistent with transcriptome.The in vivo functional tests showed that GEN-617 increased body weight,HbA1c,triglyceride and total cholesterol levels in the diabetic rats(P<0.05 or P<0.01);while all the above-mentioned levels(except triglyceride)were decreased significantly by GEN-617 combined with DGR intervention(P<0.05 or P<0.01).Conclusion:Nampt activation was one of the mechanisms about DGR regulating glycolipid metabolism.

Efficacy of Dangua Fang(丹瓜方) on endothelial cells damaged by oxidative stress

OBJECTIVE: To evaluate the protective effects of serum containing Dangua Fang( 丹 瓜 方) on vascular endothelium damaged by oxidative stress. METHODS: Five experiments were completed in this paper. In the first experiment, we found the most suitable serum containing Dangua Fang by comparing groups with different serum containing Dangua Fang. In the second experiments we analyzed Dangua Fang influencing endothelial cell viability and apoptosis and cell cycle. The third experiment on Dangua Fang intervention of mitochondrial respiratory chain. The fourth experiment on Dangua Fang intervention of mitochondrial membrane potential. And finally, on the fifth experiment we researched the mechanism of Dangua Fang improving mitochondrial function by comparing the Na~+-k~+-ATPase and peroxisome proliferator-activated receptor-gamma coactivator-1alpha(PGC-1α) in the Dangua group with the diazoxide group and Co Q+Vit C group. RESULTS: We compared the control group in the first experiments and the OD values in DZ1 group was the most significant in all intervening groups. The recipe of DZ1(5% serum containing Dangua Fang) was used in the following experiments. Compared with the control group, cell viability, cell cycle(G2 + S), cytochrome c oxidase(COX), R3 red/green, R2 red/green, R1 red/green decreased and apoptosis, succinate dehydrogenase(SDH), green(R2 + R3), Na+-k+-ATPase, PGC-1α increased in the model group. Compared with the model group, cell viability, G2+S, COX, R3 red/green, R2 red/green, R1 red/green raised and apoptosis, green(R2 + R3), Na-K-ATPase decreased in the Dangua group;G2 + S, R3 red/green, R2 red/green, R1 red/green raised and green(R2 + R3) decreased in the Co Q + Vit C group. Na-K-ATPase increased in the combined group(P < 0.05 or < 0.01). CONCLUSIONS: Dangua Fang protects oxidative stressinduced endothelial cells damaged by promotion of mitochondrial biogenesis, reduction of Na~+-K~+-ATPase activity and regulation of mitochondrial respiratory chain function restoring mitochondrial membrane potential.

Dangua Fang(丹瓜方)regulating tricarboxylic acid cycle and respiratory chain and its mechanism in diabetic rats

OBJECTIVE:To investigate the influence and possible targets of Dangua Fang(丹瓜方)on tricarboxylic acid(TCA)cycle and respiratory chain to enrich the prescription’s mechanism of effective intervention on glycolipid metabolic diseases such as type 2 diabetes.METHODS:After interventional rats were fed with high glucose and high fat diet ad libitum for 4 weeks,intraperitoneally injected streptozotocin to induce diabetic model.According to blood glucose level,28 diabetic rats were selected and continued to be fed with high glucose and high fat diet,were stratified by body weight,and divided randomly by blood glucose into Model group(was given sterile water by gastric perfusion and injected aquae pro injection intraperitoneally),Dangua group[Dangua liquor(丹瓜方液)20.5 g·kg^(-1)·d^(-1) by perfusion and aquae pro injection intraperitoneally],Inhibitor group[sterile water by perfusion and nicotinamide phosphoribosyl transferase(Nampt)specific blocker GEN-6171.25 mg/kg intraperitoneally],DanInhit group(Dangua liquor and GEN-617 synchronously).Control group were continuously fed with ordinary diet.The intervention was last for 10 weeks.Body weight(BW),liver index(LI),glycosylated hemoglobin(HbA1c),TC,TG,free fatty acids(FFA),creatinine(Cr),and A-ketoglutarate(α-KG),Iso-citric acid(ICA),oxaloacetic acid(OAA)were tested.The cytochrome C oxidase(COX)and Succinate dehydrogenase(SDH)were evaluated by Colorimetry;Nampt protein,Adenosine triphosphate(ATP)synthase(ATPs),Nicotinamide adenine dinucleotide(NAD^(+))and its reduced(NADH)in liver were measured by enzyme linked immunosorbent assay.The expressions of Nampt and mitochondrialnadhdehydrogenase-1(mt-ND1)gene in liver was assessed by real-time polymerase chain reaction.Hepatic tissue staining was also completed.RESULTS:The levels of BW,ICA,α-KG and Nampt-mRNA in the Model group are lower than that in the Normal group(P<0.05),conversely,liver weight,LI,TC,HbA1c,SDH and ATPs,mt-ND1-mRNA,and Nampt protein in the Model group are higher(P<0.01,P<0.05).Compared with Model group,the levels of ICA,Nampt-mRNA and Nampt in Dangua group are significantly increased,and FFA obviously raised(P<0.01 and P<0.05);liver weight,BW,SDH are obviously lower,and HbA1c decreased significantly(P<0.01,P<0.05).TG,FFA and Nampt protein increased in the DanInhit group,TC,TG,BW obviously increased in the Inhibitor group,but SDH is decreased in both the two groups(P<0.05,P<0.01).Compared with Dangua group,DanInhib group has the lower levels of ICA,mt-ND1-mRNA,Nampt-mRNA,and the higher level of BW,LI and HbA1c.In the Inhibitor group,ICA and Nampt protein decreased,BW and LI,HbA1c and TG increased(P<0.01 or P<0.05).Tissue staining display that,in the model group there is obvious pathologic changes ie:fibrosis,steatosis and inflammatory cell infiltration.Lesions in the Dangua group are mild,and those of Inhibitor group are more obvious than the Model group,and DanInhit group is intermediately affected compared to Dangua group and Inhibitor group.CONCLUSION:Dangua Fang increases the metabolic flux of TCA cycle and optimizes respiratory chain function by up-regulating Nampt expression.

A clinical study of Dangua Humai oral liquid on cardiovascular risk factors among patients with type 2 diabetes mellitus

Objective To study the influence of Dangua Humai oral liquid on inflammatory markers and cardiovascular prognosis among patients with high cardiovascular risks of type 2 diabetes mellitus. Methods A sample of 70 participants were randomly assigned to an experimental group(35 cases) and a control group(35 cases).

丹栝方干预糖尿病动脉粥样硬化大鼠糖脂代谢及氧化应激研究

目的探讨丹栝方对糖尿病动脉粥样硬化大鼠糖脂代谢及血清活性氧族(ROS)、胸主动脉核因子κB(nuclear factor kappa B,NF-κB)阳性表达及其mRNA表达水平的影响,以揭示其干预糖尿病慢性并发症的部分机制。方法选取自发性糖尿病GK大鼠(Goto-Kakisaki Wistar rats)40只,以含代谢抑制剂丙硫氧嘧啶的高脂饲料饲养及内皮型一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸甲酯腹腔注射,制备高脂糖尿病动脉硬化模型。将GK糖尿病成模鼠先按体重分层,再按血糖浓度根据随机数字表法分为丹栝方组[丹栝方,8mL/(kg·d)]、二甲双胍组[二甲双胍,150mg/(kg·d)]、辛伐他汀组[辛伐他汀,2mg/(kg·d)]、模型组[纯净水,8mL/(kg·d)],每组各10只;另取同龄、体重可比Wistar大鼠10只作为正常对照组(正常组)。各组均灌胃干预24周。监测空腹血糖(FBG)、体重,测定HbA1c、TC、LDL-C、HDL-C、TG、血清ROS,免疫组化法分析主动脉NF-κB,Real-time PCR检测主动脉NF-κB(P65) mRNA表达。结果正常组体重最终显著超过其他各组(P<0.01),四组GK造模组间比较,差异无统计学意义。四组GK造模鼠全程FBG都高于正常组(P<0.01,P<0.05);首次FBG、基线血糖各组间比较,差异无统计学意义(P>0.05),末次FBG二甲双胍组和丹栝方组都显著低于模型组(P<0.01)和辛伐他汀组(P<0.05)。干预24周后,四组GK造模鼠FBG、HbA1c、TC、LDL-C、HDL-C、胸主动脉NF-κB的阳性表达率,及模型组、二甲双胍组、丹栝方组胸主动脉NF-κB mRNA的表达均高于正常组(P<0.01,P<0.05);二甲双胍组、丹栝方组、辛伐他汀组TG、血清ROS,及辛伐他汀组胸主动脉NF-κB mRNA表达低于正常组(P<0.01,P<0.05)。药物干预三组FBG、TC、LDL-C、血清ROS、胸主动脉NF-κB mRNA的表达,丹栝方组、二甲双胍组胸主动脉NF-κB的阳性表达率,及丹栝方组HbA1c、TG显著低于模型组(P<0.01,P<0.05)。二甲双胍组、丹栝方组FBG低于辛伐他汀组(P<0.05)。辛伐他汀组、丹栝方组胸主动脉NF-κB mRNA的表达,及丹栝方组TC、LDL-C显著低于二甲双胍组(P<0.01)。结论丹栝方通过综合调节血糖、血脂,下调氧化应激,以发挥其防治糖尿病的慢性并发症作用。

丹瓜方对GK糖尿病-动脉粥样硬化复合模型大鼠血糖、食量和体重的影响

目的:探讨丹瓜方对高脂饲料喂养的GK糖尿病-动脉粥样硬化复合模型大鼠血糖、食量和体重的影响。方法:40只成模GK大鼠,随机分为二甲双胍组、模型对照组、丹瓜方组、辛伐他汀组,自由进食高脂饲料,并予以相应的药物干预;另设10只同龄Wistar大鼠为正常对照组,喂食普通饲料;观察大鼠血糖、HbA1c、体重、食量的变化情况。结果:丹瓜方组血糖与空白对照组比较无差异,且优于其他各模型组;丹瓜方组HbA1c与二甲双胍组比较无差别,且低于模型对照组(P<0.05);GK大鼠食量均明显减少,丹瓜方组食量低于空白对照组和模型对照组(P<0.01);GK大鼠体重均低于空白对照组,丹瓜方组体重增幅最小(P<0.05)。结论:丹瓜方在控制高脂饲料喂养的GK大鼠的血糖、体重和食量上具有明显优势。

丹瓜方对糖尿病大鼠离体心脏垂体后叶素所致心肌缺血模型的影响

目的:观察中药复方丹瓜方对高脂糖尿病大鼠离体心脏的垂体后叶素所致心肌缺血模型的影响,并探讨其可能的作用机制。方法:选取SD大鼠,随机分出正常组大鼠,其余大鼠经糖尿病造模后,随机分为模型组、糖脉康组及丹瓜方低、中、高剂量组。干预12周后采用Langendorff恒流灌流装置及加有垂体后叶素的K-H液灌注离体心脏,分别观察并记录各时间段的冠脉流量、心率及心肌收缩幅度。结果:与正常组相比,中、高剂量组第30分钟心肌收缩幅度均升高(P<0.05);与高剂量组相比,低剂量组和模型组第10分钟心率均降低(P<0.05),低剂量组、模型组及正常组第30分钟心率均降低(P<0.05);与中剂量相比,正常组及模型组第15分钟心肌收缩幅度均下降(P<0.05或P<0.01)。结论:丹瓜方对糖尿病缺血缺氧心肌有一定的保护作用。

丹瓜护脉口服液对2型糖尿病合并非酒精性脂肪性肝病患者血管内皮功能及氧化应激指标的影响

目的:探讨痰瘀同治指导下丹瓜护脉口服液对2型糖尿病(type 2 diabetes mellitus,T2DM)合并非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)患者血管内皮功能及氧化应激指标的影响。方法:选取2018年1月至2020年7月福建中医药大学附属人民医院收治的T2DM合并NAFLD患者92例,按照入院先后顺序分为对照组46例和观察组46例。对照组患者给予常规治疗,观察组患者在对照组的基础上根据痰瘀同治指导理论予以丹瓜护脉口服液,均治疗3个月。比较两组患者的疗效,治疗前后血糖[空腹血糖(FBG)、餐后2 h血糖(2 hBG)及糖化血红蛋白(HbA lc)]、血脂[总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)及低密度脂蛋白(LDL)]、肝功能[丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)及γ-谷氨酰转移酶(GGT)]、血管内皮功能[血管内皮生长因子(VEGF)、内皮素(ET-1)]及氧化应激[丙二醛(MDA)、超氧化物歧化酶(SOD)]等指标水平。结果:观察组患者的总有效率为89.13%(41/46),明显高于对照组的71.74%(33/46),差异有统计学意义(P<0.05)。治疗后,两组患者FBG、2 hBG、HbA lc、TC、TG、LDL、ALT、AST、GGT、ET-1及MDA水平较治疗前明显降低,HDL、VEGF及SOD水平明显升高,差异均有统计学意义(P<0.05);观察组患者的FBG、HbA lc、ALT、AST及GGT水平与对照组的差异均无统计学意义(P>0.05),2 hBG、TC、TG、LDL、ET-1及MDA水平明显低于对照组,HDL、VEGF及SOD水平明显高于对照组,差异均有统计学意义(P<0.05)。结论:痰瘀同治指导下使用丹瓜护脉口服液治疗,能够更好地调节T2DM合并NAFLD患者的血脂和血糖代谢,改善其血管内皮功能,减轻氧化应激反应,提高治疗效果,且无明显不良反应。

丹瓜方对糖尿病大鼠胰腺形态及GLUT4、LEP表达的影响

目的探讨丹瓜方对糖尿病大鼠胰腺形态及葡萄糖转运蛋白4(glucose transporter 4,GLUT4)、瘦素(leptin,LEP)表达的影响。方法SD大鼠按前期研究方案造模成糖尿病大鼠,随机分为正常组、模型组、二甲双胍组、常规剂量组、高剂量组。干预16周后分离获取胰腺组织并进行HE染色,检测糖尿病大鼠肌肉和脂肪组织GLUT4、LEP的表达。结果HE染色形态:正常组胰岛细胞外观呈饱满球形,细胞排列匀齐,胞质内含有大量胰岛素分泌颗粒,胰岛细胞占胰岛大部分范围;胰岛数量较多,面积较大,均衡分布并和腺泡分界清楚。与正常组比较,模型组胰岛细胞形态萎缩,排列混乱;胞质内胰岛素分泌颗粒显著减少;胰岛数量较少,面积较小,不均匀分布。与模型组比较,常规剂量组、高剂量组及二甲双胍组胰岛结构较规整,胰岛面积较大,胰岛细胞萎缩程度较轻,胰岛素分泌颗粒较多。与正常组比较,模型组和二甲双胍组大鼠肌肉GLUT4蛋白的表达量显著降低(P<0.01),常规剂量组大鼠肌肉GLUT4蛋白表达显著增加(P<0.01);模型组大鼠脂肪LEP的mRNA表达量显著降低(P<0.01)。和模型组相比,常规剂量组、高剂量组的大鼠肌肉GLUT4蛋白表达显著增加(P<0.01),二甲双胍组、常规剂量组、高剂量组大鼠脂肪LEP的mRNA表达量增加(P<0.05或P<0.01);与二甲双胍组比较,常规剂量组及高剂量组大鼠肌肉GLUT4蛋白表达显著增加(P<0.01),常规剂量组大鼠脂肪LEP的mRNA表达量显著增加(P<0.05)。结论丹瓜方可改善胰岛细胞的形态,丹瓜方可以通过调节GLUT4蛋白及LEP的mRNA表达来影响胰岛素功能达到控制血糖的目的。

HPLC法测定丹瓜护脉颗粒中芍药苷和丹酚酸B的含量

目的建立HPLC法测定丹瓜护脉颗粒中芍药苷和丹酚酸B含量的方法,为质量控制提供依据。方法HPLC法,采用Ulitamate C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈-0.1%磷酸溶液(17∶83);流速:1 mL·min^(-1);柱温:室温;检测波长:230 nm。结果芍药苷和丹酚酸B在2.5~80μg·mL^(-1)范围内线性关系良好,相关系数分别为0.9999和0.9997;平均回收率(n=9)分别为96.84%(RSD=1.26%)和99.09%(RSD=2.26%)。结论方法简便,准确,稳定好,可用于丹瓜护脉颗粒的质量控制。

丹瓜方治疗长期血糖控制不良2型糖尿病疗效观察

目的观察丹瓜方治疗顽固性高血糖、长期不能控糖达标的2型糖尿病的疗效。方法将126例长期血糖控制不良的2型糖尿病患者随机分为治疗组82例和对照组44例,对照组根据情况予胰岛素、口服降糖药单药或联合治疗,治疗组在对照组的基础上加用丹瓜方治疗,观察2组临床症状、血糖、糖化血红蛋白(HbA1C)、C肽、血液流变学、纤维蛋白原(FIB)的变化。结果与对照组相比,治疗组治疗后餐后2 h血糖、HbA1C、餐后0.5 h C肽、餐后2 h C肽、全血高切粘度、全血低切粘度、FIB、证候积分明显下降(P<0.05),但2组间空腹血糖、空腹C肽没有统计学差异;治疗组胰岛素平均用量明显小于对照组(P<0.05),但2组之间格列齐特缓释片、二甲双胍缓释片、罗格列酮钠的平均用量没有统计学差异。治疗组总有效率为92.68%,对照组为77.27%,治疗组临床疗效明显优于对照组(P<0.05)。结论丹瓜方可有效改善糖尿病患者的临床症状,有一定的降糖、改善胰岛功能、改善血粘度的作用;同时联合使用丹瓜方可减少降糖药用量。

丹瓜方对糖尿病大鼠胸主动脉超微结构影响的初步观察

目的观察祛瘀化痰、痰瘀同治复方中药——丹瓜方对糖尿病大鼠大血管超微结构的影响。方法 40只糖尿病GK大鼠(Goto-Kakisaki Wista rat)按数字表随机等分为模型组、二甲双胍组、辛伐他汀组、丹瓜方组4组,Wistar大鼠作为正常对照组。各组分别灌胃,24周后解剖取胸主动脉。透射电镜观察各组大鼠主动脉的超微结构。结果丹瓜方干预后大鼠胸主动脉内膜表面未见明显突起、增厚;内皮细胞较平滑,紧贴基层;弹力板连续、较平滑,厚薄相近;平滑肌细胞连续、完整,整体效果优于二甲双胍组。结论丹瓜方能显著改善糖尿病模型大鼠胸主动脉超微形态学变化,有明显的保护血管作用。

丹瓜护脉口服液对痰瘀型2型糖尿病合并NAFLD患者糖脂代谢、胰岛素抵抗及肝纤维化程度的疗效观察

目的:以中成药丹瓜护脉口服液干预痰瘀型2型糖尿病合并非酒精性脂肪性肝病(NAFLD)患者,观察其临床疗效。方法:选取2018年6月1日-12月31日在本科住院的105例痰瘀型T2DM合并NAFLD患者,随机分为对照组50例及治疗组55例。对照组按指南规范给予降糖、降压、降脂及抗血小板等西医治疗;治疗组在此基础上加用丹瓜护脉口服液,疗程6个月。观察两组治疗前后糖代谢(FPG、HbA1c、FINS)、脂代谢(TC、TG、HDL-C、LDL-C)、肝酶学指标(ALT、AST),计算HOMA-IR与NAFLD纤维化评分(NFS)的变化,以及脂肪肝程度的变化。结果:(1)治疗前,两组血糖、血脂、HOMA-IR、肝酶学指标等差异无统计学意义(P>0.05)。(2)治疗组治疗6个月后,组内治疗前后比较显示,糖脂代谢、肝酶指标及计算HOMA-IR、NFS也较治疗前明显下降(P<0.05),而FINS、HDL-C水平无明显变化(P>0.05)。对照组治疗6个月后,上述糖脂代谢、肝酶指标等实验室检查指标较治疗前亦有下降(P<0.05),但HOMA-IR、NFS下降并不明显(P>0.05)。(3)组间治疗后比较,治疗6个月后治疗组不仅糖脂代谢及肝酶指标低于对照组,HOMA-IR、NFS也显著低于对照组(P<0.05)。(4)治疗组脂肪肝治疗的总有效率显著高于对照组(P<0.05)。结论:丹瓜护脉口服液用于痰瘀型T2DM合并NAFLD患者在有效改善糖脂代谢的基础上,可改善胰岛素抵抗,同时降低肝损伤并显著改善脂肪肝程度,从而延缓肝纤维化进展。

丹瓜方防治糖尿病鼠氧化应激损伤研究

目的观察丹瓜方对糖尿病鼠肝组织的影响。方法将中国地鼠按计算器随机数分为空白组(KB)、模型组(MX)、低剂量中药丹瓜方组(DY)、中剂量中药丹瓜方组(ZY)、高剂量中药丹瓜方组(GY)5组。KB组饲以普通饲料,其他组予高脂高糖饲料并相应药物干预。4周后取材观测地鼠肝组织SOD、H2O2、GSH、MDA的变化。结果与MX组比较,KB组、DY组、ZY组、GY组的H2O2含量有统计学差异(P<0.05,P<0.01),GY组SOD活性升高(P<0.05),GY组GSH含量升高(P<0.01),ZY组MDA含量下降(P<0.05)。结论丹瓜方可以升高糖尿病鼠肝组织SOD、GSH水平,降低H2O2、MDA水平,丹瓜方可通过抗氧化应激途径防治糖尿病。

丹瓜方防治转基因糖尿病小鼠的氧化应激肝损伤

目的研究丹瓜方对转基因糖尿病小鼠氧化应激肝损伤的影响。方法将32只糖尿病C57BL/6J小鼠按随机数字表分为模型组、丹瓜方组、丹瓜方加吡格列酮组(联合用药组)、吡格列酮组,每组8只,喂养12周,处死,取肝脏制作匀浆,用化学荧光法测定活性氧簇(ROS)活力,比色法测定还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)活力。结果丹瓜方组、联合用药组、吡格列酮组ROS活力低于模型组,SOD值高于模型组(P<0.05);联合用药组和丹瓜方组GSH活力高于模型组(P<0.05,P<0.01),与正常组比较差异有统计学意义(P<0.05);联合用药组与丹瓜方组GSH活力比较差异无统计学意义。结论丹瓜方可升高糖尿病鼠肝组织SOD、GSH水平,降低ROS水平,通过抗氧化应激途径防治糖尿病。

丹瓜方治疗2型糖尿病合并非酒精性脂肪性肝病临床观察

目的观察丹瓜方对痰瘀型2型糖尿病合并非酒精性脂肪性肝病(NAFLD)患者肝纤维化评分(NFS)和血清肝纤维化指标的影响。方法将112例痰瘀型2型糖尿病合并NAFLD且NFS>0.676的患者用随机数字表法随机分为对照组55例、治疗组57例。对照组予健康教育、饮食控制、运动治疗、降糖和降脂治疗。治疗组在对照组干预基础上加用丹瓜方。2组均连续治疗12周。观察治疗前后2组FBG、2 h-BG、HbA1c、血脂、肝功、NFS、肝纤维化指标及证候积分的变化。结果与对照组相比,治疗组治疗后FBG、Hb A1c、AST、TC、C-IV、HA、NFS和证候积分明显降低(P<0.05),2组治疗后ALT、GGT、TG、LDL比较,差异没有统计学意义(P>0.05)。结论丹瓜方能够改善及延缓2型糖尿病合并NAFLD患者的肝纤维化的进展,同时具有改善糖脂代谢、保护肝功能的作用。

丹瓜方对痰瘀型2型糖尿病患者血清TGF-β1、ox-LDL的影响

目的观察丹瓜方对痰瘀型2型糖尿病患者血清TGF-β1、ox-LDL的影响。方法将144例痰瘀型2型糖尿病患者随机分为观察组和对照组各72例。对照组采用常规降糖、降压、调脂治疗,观察组在对照组治疗基础上联合丹瓜方治疗,疗程均为4周。观察2组治疗前后FBG、2h PG、TC、TG、LDL、TGF-β1、ox-LDL及证候积分的变化。结果观察组剔除3例,对照组剔除2例。与对照组比较,观察组治疗后2h PG、TC、LDL、TGF-β1、ox-LDL水平及证候积分明显下降(P<0.05)。结论丹瓜方可改善痰瘀型2型糖尿病患者糖脂代谢,可抑制糖尿病炎症反应并减轻血管内皮损伤,具有抗动脉粥样硬化的作用。

丹瓜方对2型糖尿病模型大鼠骨骼肌脂联素的影响

目的研究丹瓜方对2型糖尿病GK大鼠骨骼肌脂联素表达的影响。方法 40只2型糖尿病GK大鼠,随机分为二甲双胍组、对照组、丹瓜方组、辛伐他汀组,自由进食高脂饲料,并予以对应药物干预;另设10只同龄的Wistar大鼠为正常对照组,予以普通饮食。干预24周后,检测大鼠血糖、血脂水平,用酶联免疫吸附法测定GK大鼠骨骼肌脂联素表达。结果丹瓜方和二甲双胍在控制GK大鼠血糖上无显著差别;丹瓜方能明显降低CHOL、TG和LDL-c,升高HDL-c,效果明显优于辛伐他汀和二甲双胍;丹瓜方能明显促进GK大鼠骨骼肌脂联素的表达,优于二甲双胍及辛伐他汀。结论丹瓜方在降低高脂饲养的2型糖尿病GK大鼠的血糖血脂同时,能显著提高骨骼肌脂联素水平。

丹瓜护脉颗粒质量标准研究及药效评价

目的建立丹瓜护脉颗粒(DGHM)的质量标准以及其降血糖药效研究。方法采用高效液相色谱法测定DGHM中芍药苷与丹酚酸B含量;通过小鼠实验评价DGHM对肾上腺素引起的急性高血糖模型小鼠的降糖效果;通过小鼠实验评价DGHM对脲佐链霉素(STZ)引起的糖尿病模型小鼠的降糖作用。结果芍药苷和丹酚酸B的平均加样回收率和RSD分别为97.63%(1.33%),95.00%(2.43%);DGHM对肾上腺素高血糖小鼠均有降血糖作用,与空白组比较,差异有统计学意义(P<0.05);DGHM对STZ导致的高血糖小鼠具有一定效果,低于模型对照组(P<0.05),糖耐量显著提高。结论本方法准确、快捷,可用于丹瓜护脉制剂的质量控制;丹瓜护脉制剂具有抗糖尿病与降血糖功效。