本文關(guān)鍵詞： Wistar大鼠 多西紫杉醇 化療 胰島 血糖　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:隨著人口老齡化的加劇,人們生活方式的改變,糖尿病的患病率呈上升態(tài)勢,同時(shí),癌癥患病率亦急劇升高,而治愈率較低,嚴(yán)重威脅了人類健康。由于手術(shù)和放療的局限性,化療逐漸成為一種臨床常用的癌癥治療手段。近來化療藥物在骨髓抑制、消化道反應(yīng)、肝腎功能損害、過敏反應(yīng)等方面的副作用已被逐漸揭示,另外有研究發(fā)現(xiàn),化療藥物對機(jī)體的糖代謝方面包括胰島功能改變、糖代謝異常有一定影響,甚至誘導(dǎo)糖尿病發(fā)生。紫杉類藥物(紫杉醇和多西紫杉醇)已被廣泛用于卵巢癌、乳腺癌、肺癌等癌癥的治療。研究表明紫杉類藥物降低腫瘤患者死亡率,也影響機(jī)體的糖代謝,而相關(guān)機(jī)制尚未完全闡明。本研究對正常Wistar大鼠及2型糖尿病大鼠腹腔注射多西紫杉醇,探討多西紫杉醇對正常大鼠和糖尿病大鼠血糖的影響;探討多西紫杉醇對大鼠胰島功能及組織結(jié)構(gòu)的影響;分離并體外培養(yǎng)正常大鼠的胰島,觀察多西紫杉醇對胰島的直接影響,為有或無糖尿病的癌癥患者的個(gè)體化化療的順利進(jìn)行提供理論依據(jù)。方法:將6周齡雌性Wistar大鼠隨機(jī)分為兩組,分別給予基礎(chǔ)飼料(NC組)或高脂飼料喂養(yǎng)(HF組),8周后行口服葡萄糖耐量實(shí)驗(yàn),HF組證實(shí)出現(xiàn)胰島素抵抗時(shí),給予腹腔注射鏈脲佐菌素30mg/kg,72h后測血糖值,以隨機(jī)血糖≥16.7mmol/L者證實(shí)為2型糖尿病大鼠。將正常大鼠及2型糖尿病大鼠隨機(jī)分組,正常對照組(NC組)、糖尿病對照組(DM組)給予生理鹽水,地塞米松組(NCD組)、糖尿病地塞米松組(DMD組)給予地塞米松,多西紫杉醇組(NCT組)、糖尿病多西紫杉醇組(DMT組)給予多西紫杉醇干預(yù)1周期或4周期,記錄大鼠體重、進(jìn)水量、進(jìn)食量、攝入熱量。腹腔注射STZ前后、多西紫杉醇干預(yù)前后行口服葡萄糖耐量試驗(yàn)及C-肽釋放試驗(yàn),測定血糖、胰島素、C-肽、總膽固醇(TC)、甘油三酯(TG)、谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST),取血結(jié)束后處死動(dòng)物,留取胰腺、肝臟組織檢查組織病理學(xué)變化。采用逆行灌注法分離正常大鼠胰腺,膠原酶消化,應(yīng)用Ficoll-400密度梯度離心法純化胰島,于含不同濃度多西紫杉醇的RPMI1640培養(yǎng)液(含10%胎牛血清)中,分別培養(yǎng)不同時(shí)間。后分別置于含5.6mmol/L、16.7mmol/L葡萄糖的DMEM培養(yǎng)基中培養(yǎng)1h,吸取上清液,應(yīng)用ELISA法測定胰島素水平,吸附離心法提取胰島DNA,紫外線光吸收度法測定數(shù)值。統(tǒng)計(jì)學(xué)處理采用SPSS21.0軟件,計(jì)量資料采用均數(shù)±標(biāo)準(zhǔn)差(？)表示。結(jié)果:1正常對照組大鼠(NC組)與高脂組大鼠(HF組)指標(biāo)比較1.1進(jìn)水、進(jìn)食量、攝入熱量比較實(shí)驗(yàn)前8周NC組與HF組大鼠進(jìn)水、進(jìn)食量無統(tǒng)計(jì)學(xué)差異,HF組攝入熱量明顯高于NC組,有統(tǒng)計(jì)學(xué)差異(P0.01)。1.2體重比較實(shí)驗(yàn)開始時(shí)各組大鼠體重?zé)o統(tǒng)計(jì)學(xué)差異,喂養(yǎng)8周后,HF組大鼠體重明顯高于NC組,有統(tǒng)計(jì)學(xué)差異(P0.01)。1.3口服葡萄糖耐量試驗(yàn)、C-肽釋放試驗(yàn)及HOMA-IR、ISI、△I/△G比較HF組大鼠血糖達(dá)高峰時(shí)間后延,空腹及葡萄糖負(fù)荷后30min、60min、120min血糖、胰島素、C-肽水平均高于NC組,有統(tǒng)計(jì)學(xué)差異(P0.01)。HF組大鼠HOMA-IR高于NC組,HF組大鼠ISI、△I/△G低于NC組,有統(tǒng)計(jì)學(xué)意義(P0.01)。2正常對照組大鼠(NC組)與糖尿病組大鼠(DM組)指標(biāo)比較2.1進(jìn)水、進(jìn)食量、攝入熱量比較DM組大鼠進(jìn)水、進(jìn)食量、攝入熱量明顯高于NC組,有統(tǒng)計(jì)學(xué)意義(P0.01);NCD1組、NCT1組與DMD組、DMT組大鼠進(jìn)水、進(jìn)食量、攝入熱量較前降低,有統(tǒng)計(jì)學(xué)意義(P0.01);NCD4組、NCT4組進(jìn)水、進(jìn)食量、攝入熱量較NCD1、NCT1組進(jìn)一步降低,有統(tǒng)計(jì)學(xué)意義(P0.01)。2.2體重比較NCT4組大鼠較NC4組大鼠、NCD4組體重明顯降低,有統(tǒng)計(jì)學(xué)差異(P0.01);DMT組、DMD組大鼠較DM組體重下降明顯,有統(tǒng)計(jì)學(xué)意義(P0.05)。較DM組大鼠血糖達(dá)高峰時(shí)間后延,空腹及葡萄糖負(fù)荷后30min、60min、120min血糖均較NC組升高,有統(tǒng)計(jì)學(xué)差異(P0.01);DM組大鼠空腹及葡萄糖負(fù)荷后30min、60min、120min胰島素、C-肽水平均較NC組降低,有統(tǒng)計(jì)學(xué)差異(P0.01)。DM組大鼠HOMA-IR高于NC組,有統(tǒng)計(jì)學(xué)差異(P0.01);DM組大鼠ISI、△I/△G低于NC組,有統(tǒng)計(jì)學(xué)差異(P0.01)。2.3口服葡萄糖耐量試驗(yàn)、C-肽釋放試驗(yàn)及HOMA-IR、ISI、△I/△G比NCT1組大鼠葡萄糖負(fù)荷后60min血糖比NC1組、NCD1組升高,有統(tǒng)計(jì)學(xué)意義(P0.01);NC1組、NCD1組、NCT1組大鼠胰島素、C-肽、HOMA-IR、ISI、△I/△G比較無統(tǒng)計(jì)學(xué)差異。NCT4組大鼠血糖高峰后延,空腹及葡萄糖負(fù)荷后30min、60min、120min血糖高于NC4組、NCD4組、NCT1組,有統(tǒng)計(jì)學(xué)意義(P0.01);NCT4組大鼠空腹及葡萄糖負(fù)荷后30min、60min、120min胰島素、C-肽分泌較NC4組、NCD4組降低,有統(tǒng)計(jì)學(xué)意義(P0.05);NCT4組大鼠葡萄糖負(fù)荷后30min胰島素分泌較NCT1組降低,有統(tǒng)計(jì)學(xué)意義(P0.01);NCT4組大鼠空腹及葡萄糖負(fù)荷后30min、60min、120min C-肽分泌較NCT1組降低,有統(tǒng)計(jì)學(xué)意義(P0.01);NCT4組大鼠HOMA-IR較NC4組、NCD4組、NCT1組大鼠升高,有統(tǒng)計(jì)學(xué)意義(P0.01);NCT4組大鼠ISI、△I/△G低于NC4組、NCD4組、NCT1組,有統(tǒng)計(jì)學(xué)差異(P0.05)。DMT組大鼠空腹及葡萄糖負(fù)荷后30min、60min、120min血糖較DM組、DMD組、NCT1組升高,有統(tǒng)計(jì)學(xué)差異(P0.01),DMT組空腹及葡萄糖負(fù)荷后30min、60min、120min C-肽分泌較DM組、NCT1組降低,有統(tǒng)計(jì)學(xué)意義(P0.01);DMT組空腹及葡萄糖負(fù)荷后30min C-肽分泌較DMD組降低,有統(tǒng)計(jì)學(xué)差異(P0.01)。2.4 TC、TG、ALT、AST比較NCT1組大鼠TC、TG高于NC1組,有統(tǒng)計(jì)學(xué)意義(P0.05)。NCT4組大鼠TC、TG、ALT、AST明顯高于NC1組、NCD4組、NCT1組大鼠,有統(tǒng)計(jì)學(xué)意義(P0.01);DMT組大鼠TC、TG、ALT、AST高于DM組、DMD組,有統(tǒng)計(jì)學(xué)意義(P0.05)。3胰腺、肝臟重量比較NC1組、NCD1組、NCT1組大鼠胰腺、肝臟重量比較無統(tǒng)計(jì)學(xué)差異。NCT4組胰腺、肝臟重量較NC4組、NCD4組下降,有統(tǒng)計(jì)學(xué)意義(P0.01);DMT組大鼠胰腺、肝臟重量低于DM組、DMD組,有統(tǒng)計(jì)學(xué)意義(P0.01)。4胰腺、肝臟組織形態(tài)學(xué)變化NC組大鼠胰腺胰島數(shù)量較多,細(xì)胞排列緊密,細(xì)胞核大而圓,胞漿豐富;NCT4組大鼠胰島細(xì)胞數(shù)減少;DM組大鼠胰島結(jié)構(gòu)破壞,胰島細(xì)胞數(shù)減少;DMT組大鼠胰島內(nèi)細(xì)胞數(shù)量較DM大鼠明顯減少,排列雜亂。NC組肝細(xì)胞排列規(guī)整,肝小葉規(guī)則,細(xì)胞核居中;NCT4組肝臟可見少量肝細(xì)胞空泡變性,肝竇充血、擴(kuò)張。DM組大鼠大量肝細(xì)胞空泡變性,可見脂質(zhì)浸潤;DMT組大鼠大量脂質(zhì)浸潤,肝竇充血、擴(kuò)張明顯。5體外胰島培養(yǎng)胰島素分泌比較以5.6mmol/L葡萄糖刺激時(shí),0.001ug/ml、0.005ug/ml多西紫杉醇處理胰島后胰島素分泌無明顯差異;0.01ug/ml多西紫杉醇處理1h、3h、6h、12h、24h減少胰島素分泌,有統(tǒng)計(jì)學(xué)意義(P0.01);0.01ug/ml多西紫杉醇處理6h、12h、24h較處理1h減少胰島素分泌,有統(tǒng)計(jì)學(xué)意義(P0.05)。以16.7mmol/L葡萄糖刺激時(shí),0.005ug/ml多西紫杉醇處理12h、24h胰島素分泌較未經(jīng)多西紫杉醇處理的減少,有統(tǒng)計(jì)學(xué)意義(P0.01);0.01ug/ml多西紫杉醇處理1h、3h、6h、12h、24h胰島素分泌較0 ug/ml、0.001 ug/ml、0.005 ug/ml多西紫杉醇處理的胰島素分泌減少,有統(tǒng)計(jì)學(xué)意義(P0.01)。0.005ug/ml多西紫杉醇處理12h、24h較1h SI值減低,有統(tǒng)計(jì)學(xué)意義(P0.01);0.01ug/ml多西紫杉醇組處理1h、3h、6h、12h、24h SI值降低,有統(tǒng)計(jì)學(xué)意義(P0.01);0.01ug/ml多西紫杉醇處理3h、6h、12h、24h較1h SI值減低,有統(tǒng)計(jì)學(xué)意義(P0.01)。結(jié)論:1應(yīng)用4周期多西紫杉醇可使正常大鼠胰島素分泌減少,胰島素抵抗增強(qiáng),血糖升高。2應(yīng)用1周期多西紫杉醇可進(jìn)一步損傷糖尿病大鼠胰島功能,升高血糖。3多西紫杉醇可使大鼠谷丙轉(zhuǎn)氨酶、谷草轉(zhuǎn)氨酶升高,損傷肝臟,間接導(dǎo)致血糖升高。4多西紫杉醇可使大鼠總膽固醇、甘油三酯升高,影響胰島素分泌,加強(qiáng)胰島素抵抗,進(jìn)一步升高血糖。5體外實(shí)驗(yàn)表明隨著多西紫杉醇濃度的增加,胰島素分泌逐漸減少。
[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(qiáng)HOMA-IR,ISI,鈻矷/鈻矴姣旇緝HF緇勫ぇ榧犺緋栬揪楂樺嘲鏃墮棿鍚庡歡,絀鴻吂鍙?qiáng)钁¤悇绯栬礋鑽峰悾?

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