多西紫杉醇对Wistar大鼠和糖尿病大鼠糖耐量及胰岛胰岛素分泌功能的影响

发布时间：2018-02-24 22:28

  本文关键词： Wistar大鼠 多西紫杉醇 化疗 胰岛 血糖　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:随着人口老龄化的加剧,人们生活方式的改变,糖尿病的患病率呈上升态势,同时,癌症患病率亦急剧升高,而治愈率较低,严重威胁了人类健康。由于手术和放疗的局限性,化疗逐渐成为一种临床常用的癌症治疗手段。近来化疗药物在骨髓抑制、消化道反应、肝肾功能损害、过敏反应等方面的副作用已被逐渐揭示,另外有研究发现,化疗药物对机体的糖代谢方面包括胰岛功能改变、糖代谢异常有一定影响,甚至诱导糖尿病发生。紫杉类药物(紫杉醇和多西紫杉醇)已被广泛用于卵巢癌、乳腺癌、肺癌等癌症的治疗。研究表明紫杉类药物降低肿瘤患者死亡率,也影响机体的糖代谢,而相关机制尚未完全阐明。本研究对正常Wistar大鼠及2型糖尿病大鼠腹腔注射多西紫杉醇,探讨多西紫杉醇对正常大鼠和糖尿病大鼠血糖的影响;探讨多西紫杉醇对大鼠胰岛功能及组织结构的影响;分离并体外培养正常大鼠的胰岛,观察多西紫杉醇对胰岛的直接影响,为有或无糖尿病的癌症患者的个体化化疗的顺利进行提供理论依据。方法:将6周龄雌性Wistar大鼠随机分为两组,分别给予基础饲料(NC组)或高脂饲料喂养(HF组),8周后行口服葡萄糖耐量实验,HF组证实出现胰岛素抵抗时,给予腹腔注射链脲佐菌素30mg/kg,72h后测血糖值,以随机血糖≥16.7mmol/L者证实为2型糖尿病大鼠。将正常大鼠及2型糖尿病大鼠随机分组,正常对照组(NC组)、糖尿病对照组(DM组)给予生理盐水,地塞米松组(NCD组)、糖尿病地塞米松组(DMD组)给予地塞米松,多西紫杉醇组(NCT组)、糖尿病多西紫杉醇组(DMT组)给予多西紫杉醇干预1周期或4周期,记录大鼠体重、进水量、进食量、摄入热量。腹腔注射STZ前后、多西紫杉醇干预前后行口服葡萄糖耐量试验及C-肽释放试验,测定血糖、胰岛素、C-肽、总胆固醇(TC)、甘油三酯(TG)、谷丙转氨酶(ALT)、谷草转氨酶(AST),取血结束后处死动物,留取胰腺、肝脏组织检查组织病理学变化。采用逆行灌注法分离正常大鼠胰腺,胶原酶消化,应用Ficoll-400密度梯度离心法纯化胰岛,于含不同浓度多西紫杉醇的RPMI1640培养液(含10%胎牛血清)中,分别培养不同时间。后分别置于含5.6mmol/L、16.7mmol/L葡萄糖的DMEM培养基中培养1h,吸取上清液,应用ELISA法测定胰岛素水平,吸附离心法提取胰岛DNA,紫外线光吸收度法测定数值。统计学处理采用SPSS21.0软件,计量资料采用均数±标准差(？)表示。结果:1正常对照组大鼠(NC组)与高脂组大鼠(HF组)指标比较1.1进水、进食量、摄入热量比较实验前8周NC组与HF组大鼠进水、进食量无统计学差异,HF组摄入热量明显高于NC组,有统计学差异(P0.01)。1.2体重比较实验开始时各组大鼠体重无统计学差异,喂养8周后,HF组大鼠体重明显高于NC组,有统计学差异(P0.01)。1.3口服葡萄糖耐量试验、C-肽释放试验及HOMA-IR、ISI、△I/△G比较HF组大鼠血糖达高峰时间后延,空腹及葡萄糖负荷后30min、60min、120min血糖、胰岛素、C-肽水平均高于NC组,有统计学差异(P0.01)。HF组大鼠HOMA-IR高于NC组,HF组大鼠ISI、△I/△G低于NC组,有统计学意义(P0.01)。2正常对照组大鼠(NC组)与糖尿病组大鼠(DM组)指标比较2.1进水、进食量、摄入热量比较DM组大鼠进水、进食量、摄入热量明显高于NC组,有统计学意义(P0.01);NCD1组、NCT1组与DMD组、DMT组大鼠进水、进食量、摄入热量较前降低,有统计学意义(P0.01);NCD4组、NCT4组进水、进食量、摄入热量较NCD1、NCT1组进一步降低,有统计学意义(P0.01)。2.2体重比较NCT4组大鼠较NC4组大鼠、NCD4组体重明显降低,有统计学差异(P0.01);DMT组、DMD组大鼠较DM组体重下降明显,有统计学意义(P0.05)。较DM组大鼠血糖达高峰时间后延,空腹及葡萄糖负荷后30min、60min、120min血糖均较NC组升高,有统计学差异(P0.01);DM组大鼠空腹及葡萄糖负荷后30min、60min、120min胰岛素、C-肽水平均较NC组降低,有统计学差异(P0.01)。DM组大鼠HOMA-IR高于NC组,有统计学差异(P0.01);DM组大鼠ISI、△I/△G低于NC组,有统计学差异(P0.01)。2.3口服葡萄糖耐量试验、C-肽释放试验及HOMA-IR、ISI、△I/△G比NCT1组大鼠葡萄糖负荷后60min血糖比NC1组、NCD1组升高,有统计学意义(P0.01);NC1组、NCD1组、NCT1组大鼠胰岛素、C-肽、HOMA-IR、ISI、△I/△G比较无统计学差异。NCT4组大鼠血糖高峰后延,空腹及葡萄糖负荷后30min、60min、120min血糖高于NC4组、NCD4组、NCT1组,有统计学意义(P0.01);NCT4组大鼠空腹及葡萄糖负荷后30min、60min、120min胰岛素、C-肽分泌较NC4组、NCD4组降低,有统计学意义(P0.05);NCT4组大鼠葡萄糖负荷后30min胰岛素分泌较NCT1组降低,有统计学意义(P0.01);NCT4组大鼠空腹及葡萄糖负荷后30min、60min、120min C-肽分泌较NCT1组降低,有统计学意义(P0.01);NCT4组大鼠HOMA-IR较NC4组、NCD4组、NCT1组大鼠升高,有统计学意义(P0.01);NCT4组大鼠ISI、△I/△G低于NC4组、NCD4组、NCT1组,有统计学差异(P0.05)。DMT组大鼠空腹及葡萄糖负荷后30min、60min、120min血糖较DM组、DMD组、NCT1组升高,有统计学差异(P0.01),DMT组空腹及葡萄糖负荷后30min、60min、120min C-肽分泌较DM组、NCT1组降低,有统计学意义(P0.01);DMT组空腹及葡萄糖负荷后30min C-肽分泌较DMD组降低,有统计学差异(P0.01)。2.4 TC、TG、ALT、AST比较NCT1组大鼠TC、TG高于NC1组,有统计学意义(P0.05)。NCT4组大鼠TC、TG、ALT、AST明显高于NC1组、NCD4组、NCT1组大鼠,有统计学意义(P0.01);DMT组大鼠TC、TG、ALT、AST高于DM组、DMD组,有统计学意义(P0.05)。3胰腺、肝脏重量比较NC1组、NCD1组、NCT1组大鼠胰腺、肝脏重量比较无统计学差异。NCT4组胰腺、肝脏重量较NC4组、NCD4组下降,有统计学意义(P0.01);DMT组大鼠胰腺、肝脏重量低于DM组、DMD组,有统计学意义(P0.01)。4胰腺、肝脏组织形态学变化NC组大鼠胰腺胰岛数量较多,细胞排列紧密,细胞核大而圆,胞浆丰富;NCT4组大鼠胰岛细胞数减少;DM组大鼠胰岛结构破坏,胰岛细胞数减少;DMT组大鼠胰岛内细胞数量较DM大鼠明显减少,排列杂乱。NC组肝细胞排列规整,肝小叶规则,细胞核居中;NCT4组肝脏可见少量肝细胞空泡变性,肝窦充血、扩张。DM组大鼠大量肝细胞空泡变性,可见脂质浸润;DMT组大鼠大量脂质浸润,肝窦充血、扩张明显。5体外胰岛培养胰岛素分泌比较以5.6mmol/L葡萄糖刺激时,0.001ug/ml、0.005ug/ml多西紫杉醇处理胰岛后胰岛素分泌无明显差异;0.01ug/ml多西紫杉醇处理1h、3h、6h、12h、24h减少胰岛素分泌,有统计学意义(P0.01);0.01ug/ml多西紫杉醇处理6h、12h、24h较处理1h减少胰岛素分泌,有统计学意义(P0.05)。以16.7mmol/L葡萄糖刺激时,0.005ug/ml多西紫杉醇处理12h、24h胰岛素分泌较未经多西紫杉醇处理的减少,有统计学意义(P0.01);0.01ug/ml多西紫杉醇处理1h、3h、6h、12h、24h胰岛素分泌较0 ug/ml、0.001 ug/ml、0.005 ug/ml多西紫杉醇处理的胰岛素分泌减少,有统计学意义(P0.01)。0.005ug/ml多西紫杉醇处理12h、24h较1h SI值减低,有统计学意义(P0.01);0.01ug/ml多西紫杉醇组处理1h、3h、6h、12h、24h SI值降低,有统计学意义(P0.01);0.01ug/ml多西紫杉醇处理3h、6h、12h、24h较1h SI值减低,有统计学意义(P0.01)。结论:1应用4周期多西紫杉醇可使正常大鼠胰岛素分泌减少,胰岛素抵抗增强,血糖升高。2应用1周期多西紫杉醇可进一步损伤糖尿病大鼠胰岛功能,升高血糖。3多西紫杉醇可使大鼠谷丙转氨酶、谷草转氨酶升高,损伤肝脏,间接导致血糖升高。4多西紫杉醇可使大鼠总胆固醇、甘油三酯升高,影响胰岛素分泌,加强胰岛素抵抗,进一步升高血糖。5体外实验表明随着多西紫杉醇浓度的增加,胰岛素分泌逐渐减少。
[Abstract]:Objective: with the aging of population, the change of people's lifestyle, the prevalence of diabetes is on the rise, at the same time, cancer rates also increased dramatically, and the cure rate is low, a serious threat to human health. Due to the limitation of the surgery and radiotherapy, chemotherapy has become a common clinical cancer treatment method recently. Chemotherapy drugs in the bone marrow suppression, gastrointestinal reaction, liver and kidney dysfunction, allergy and other aspects of the side effect has been gradually revealed, other studies have found that chemotherapeutic drugs on glucose metabolism in the body including islet function change, has certain influence on abnormal glucose metabolism, and even induce diabetes. Taxanes (paclitaxel and docetaxel) has been widely used in the treatment of ovarian cancer, breast cancer, lung cancer and other cancers. The results indicate that the taxanes reduce mortality in patients with cancer, also affect glucose metabolism in the machine body, and related machine The system has not been fully elucidated. The study of intraperitoneal injection of Wistar normal rats and type 2 diabetic rats to investigate the effects on blood glucose of docetaxel and docetaxel in normal rats and diabetic rats; to investigate the effect of docetaxel on islet function and tissue structure of rat islet; isolated and cultured normal rat, observed the direct effects of docetaxel on islet, provide a theoretical basis for individual chemotherapy with or without diabetes cancer patients smoothly. Methods: 6 week old female Wistar rats were randomly divided into two groups, were given a basal diet (NC group) or high-fat diet (HF group), 8 weeks after oral glucose tolerance test group HF, confirmed the appearance of insulin resistance, given intraperitoneal injection of streptozotocin 30mg/kg, 72h measured the blood glucose, blood glucose at random was larger than 16.7mmol/L confirmed in type 2 diabetic rats. The normal rats and type 2 diabetes Diabetic rats were randomly divided into normal control group (NC group), diabetic control group (DM group) received saline, dexamethasone group (NCD group), diabetic group (DMD group) and dexamethasone treated with dexamethasone, docetaxel group (NCT group), diabetic group (DMT group) received docetaxel docetaxel intervention for 1 or 4 cycles cycle, record the weight, water intake, food intake and calorie intake. After intraperitoneal injection of STZ and docetaxel before intervention after oral glucose tolerance test and C- peptide release test, blood glucose, insulin, C- peptide, total cholesterol (TC), triglyceride (TG), alanine aminotransferase, aspartate aminotransferase (ALT) (AST), blood samples were sacrificed after the animal, leaving the pancreas, liver histology pathological examination. The retrograde perfusion separation of normal rat pancreas, collagenase digestion, purification of islets using Ficoll-400 density gradient centrifugation in containing different concentrations of Dorsey Paclitaxel RPMI1640 medium (containing 10% fetal bovine serum), respectively. After cultured for different time were treated with 5.6mmol/L, 16.7mmol/L glucose DMEM culture supernatant was 1h in culture medium, determination of insulin levels by ELISA method. The extraction of islet DNA adsorption centrifugal method, ultraviolet light absorption method for the determination of numerical software SPSS21.0. By statistical processing, measurement data using the mean and standard deviation (?). Results: 1 rats in normal control group (NC group) and high fat group rats (HF group) compared 1.1 water intake and calorie intake compared with those before 8 weeks of NC group and HF group rats with water feeding no significant difference between the amount of calories, HF group was significantly higher than NC group, there were significant differences (P0.01) to.1.2 weight the body weight of the rats no significant difference after 8 weeks of feeding, the weight of rats in HF group were significantly higher than that of NC group, there was significant difference (P0.01).1.3 oral glucose 钀勭硸鑰愰噺璇曢獙,C-鑲介噴鏀捐瘯楠屽強HOMA-IR,ISI,鈻矷/鈻矴姣旇緝HF缁勫ぇ榧犺��绯栬揪楂樺嘲鏃堕棿鍚庡欢,绌鸿吂鍙婅憽钀勭硸璐熻嵎鍚，

本文编号：1531994