楊鑫 黃波 劉永林
[摘要]目的 觀察復(fù)方中藥組分對(duì)糖尿病的防治作用,并探討其防治機(jī)制。方法 腹腔注射鏈脲佐菌素(給藥劑量為55 mg/kg體重)誘導(dǎo)大鼠糖尿病模型,造模成功后將其隨機(jī)分為空白對(duì)照組、模型對(duì)照組、藥物干預(yù)組(中藥組分干預(yù)小劑量組、中藥組分干預(yù)大劑量組),共40只,每組10只。藥物干預(yù)組分別用大劑量和小劑量復(fù)方中藥組分進(jìn)行藥物干預(yù),空白對(duì)照組和模型對(duì)照組每天按10 ml/kg體重標(biāo)準(zhǔn)以生理鹽水灌胃,觀察空腹血糖、糖耐量及胰腺的超氧化物歧化酶(SOD)活力水平和丙二醛(MDA)含量。結(jié)果 模型對(duì)照組的血糖水平顯著高于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.001);藥物干預(yù)組0.0、0.5、1.0、2.0 h的血糖水平明顯低于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。模型對(duì)照組的糖耐量水平明顯低于空白對(duì)照組,復(fù)方中藥組分干預(yù)大劑量組一定時(shí)間內(nèi)的糖耐量水平明顯高于模型對(duì)照組。模型對(duì)照組的SOD水平明顯低于空白對(duì)照組,MDA水平明顯高于空白對(duì)照組;中藥組分干預(yù)大劑量組的SOD水平明顯高于模型對(duì)照組,MDA水平明顯低于模型對(duì)照組,差異均有統(tǒng)計(jì)學(xué)意義(P<0.01)。中藥組分干預(yù)小劑量組的SOD水平高于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。中藥組分干預(yù)小劑量組的MDA水平明顯低于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.01)。結(jié)論 復(fù)方中藥組分對(duì)糖尿病有防治作用。
[關(guān)鍵詞]復(fù)方中藥組分;抗氧化;糖耐量
[中圖分類號(hào)] R587.1 [文獻(xiàn)標(biāo)識(shí)碼] A [文章編號(hào)] 1674-4721(2019)4(c)-0045-03
Influence of compound Chinese medicine components on glucose tolerance in streptozotocin-induced diabetic rats
YANG Xin HUANG Bo LIU Yong-lin FENG Yao LIU Ming-hui ZHANG He SONG Guang-yi LIU Wen-yan▲
Liaoning Institute of Basic Medicine, Liaoning Province, Shenyang 110101, China
[Abstract] Objective To observe the preventive and therapeutic effects of compound Chinese medicine components on diabetes mellitus (DM), and to explore its prevention and treatment mechanism. Methods The rat model of DM was induced by intraperitoneal injection of streptozotocin administered at a dose of 55 mg/kg. After a successful modeling, a total of 40 rats were randomly assigned into the blank control group, the model control group and the drug intervention group (the high-dose drug intervention group, the low-dose drug intervention group), 10 cases in each group. The drug intervention group was treated with high- and low-dose of compound Chinese medicine components, the blank control group and the model control group were intragastrically administered with normal saline per day according to the weight standard of 10 ml/kg. The levels of fasting blood glucose (FBG), glucose tolerance, superoxide dismutase (SOD) activity and malondialdehyde (MDA) in pancreas were observed. Results The level of blood glucose of the model control group was significantly higher than that of the blank control group (P<0.001). The levels of blood glucose at 0.0, 0.5, 1.0, 2.0 h of the drug intervention group were significantly lower than those of the model control group (P<0.05). The levels of glucose tolerance of the model control group were significantly lower than those of the blank control group, the levels of glucose tolerance for some time of the compound Chinese medicine component intervention group in a large dose administration were significantly higher than those of the model control group. The level of SOD of the model control group was significantly lower than that of the blank control group, and the MDA of the model control group was higher than that of the blank control group, the level of SOD of the high-dose intervention group was significantly higher than that of the model control group, the level of MDA of the high-dose drug intervention group was greatly lower than that of the model control group (P<0.01). The level of SOD of the low-dose drug intervention group was higher than that of the model control group, with statistical difference (P<0.05). The MDA in the low-dose drug intervention group was much lower than that in the model control group, with statistical significance (P<0.01). Conclusion Compound Chinese herbal medicines have preventive and therapeutic effects on diabetes mellitus.
[Key words] Compound Chinese medicine components; Antioxidant; Glucose tolerance
糖尿病是與多因素相關(guān)的慢性全身性疾病,中藥及其有效組分在糖尿病及其并發(fā)癥的防治過程中有獨(dú)特的優(yōu)勢。目前已經(jīng)確認(rèn)有60多種中藥對(duì)糖尿病具有防治作用,這些藥物或具有清熱解毒功效、或具有益氣養(yǎng)陰功效、或具有活血化瘀功效,其有效成分主要為多糖、苷類、酮類和生物堿等[1-2]。本實(shí)驗(yàn)對(duì)鏈尿菌素糖尿病大鼠進(jìn)行復(fù)方中藥組分藥物干預(yù),旨在驗(yàn)證其對(duì)糖尿病的治療效果,進(jìn)而探討其治療機(jī)制。
1材料與方法
1.1材料與動(dòng)物
選取40只SD大鼠,SPF級(jí),8周齡,雌雄各半,體重為(200±10)g[許可證號(hào):SCXK(遼)2010-0001,遼寧長生生物]。藥物及試劑:鏈脲佐菌素(Streptozotocin,STZ)是Adamas Reagent CO.Ltd產(chǎn)品(貨號(hào):CFHF-33231A,批號(hào):644548120509,規(guī)格:25 g,95+);三七皂苷、麥冬、黃芪等中藥多糖組分購自北京普惠天悅公司;超氧化物歧化酶(SOD)(WST-1法)和丙二醛(MDA)(TBA法)測定試劑盒購自南京建成生物工程研究所。儀器:SXT-1型血糖儀(長沙三諾)及血糖試紙(批號(hào):1109NW);低速離心機(jī)(型號(hào)LDZ5-2,北京京立);酶標(biāo)儀(型號(hào)318C,上海三科);紫外-可見分光光度計(jì)(型號(hào)SPECORD plus200,德國耶拿)[1-3]。
1.2方法
1.2.1誘導(dǎo)大鼠糖尿病模型 大鼠適應(yīng)性飼養(yǎng)1周,禁食12 h按55 mg/kg體重一次性腹腔注射鏈脲佐菌素的枸櫞酸緩沖液(臨用前配置,STZ的濃度12.2 mg/ml,枸櫞酸緩沖液濃度為0.01 mol/L,pH值為5.0),1周后取尾尖靜脈血檢測血糖,血糖水平超過16.0 mmol/L者為造模成功,分籠備用。
1.2.2分組 給藥正常大鼠10只(雌雄各半),按55 mg/kg體重標(biāo)準(zhǔn)一次性腹腔注射枸櫞酸緩沖液作為空白對(duì)照組。將糖尿病鼠隨機(jī)分為模型對(duì)照組、中藥組分干預(yù)大劑量組、中藥組分干預(yù)小劑量組(10只/組、雌雄各半)??瞻讓?duì)照組和模型對(duì)照組每天按10 ml/kg體重標(biāo)準(zhǔn)以生理鹽水灌胃;中藥干預(yù)組的大、小劑量組每天分別按復(fù)方中藥組分500 mg/kg體重、50 mg/kg體重標(biāo)準(zhǔn)灌胃。
1.2.3指標(biāo)檢測 連續(xù)藥物干預(yù)10周后空腹12 h,取尾靜脈血檢測血糖,再以25%葡萄糖(5 g/kg)灌胃,檢測其0.5、1.0、2.0、3.0 h的血糖,做糖耐量實(shí)驗(yàn)。繼續(xù)藥物干預(yù)2周后處死各組大鼠,取胰腺制備10%組織液(3000 r/min低溫離心15 min),檢測SOD、MDA。指標(biāo)檢測按照試劑盒說明書進(jìn)行操作[4-8]。
1.3統(tǒng)計(jì)分析
采用SPSS 12.0統(tǒng)計(jì)學(xué)軟件進(jìn)行數(shù)據(jù)分析,計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示,采用t檢驗(yàn),計(jì)數(shù)資料以率(%)表示,采用χ2檢驗(yàn),以P<0.05為差異有統(tǒng)計(jì)學(xué)意義。
2結(jié)果
2.1各組不同時(shí)間段血糖水平的比較
模型對(duì)照組0.0、0.5、1.0、2.0、3.0 h的血糖水平顯著高于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.001);藥物干預(yù)組(復(fù)方中藥組分干預(yù)大劑量組、復(fù)方中藥組分干預(yù)小劑量組)0.0、0.5、1.0、2.0 h的血糖水平明顯低于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)(表1)。提示,模型對(duì)照組的糖耐量明顯低于空白對(duì)照組,復(fù)方中藥組分干預(yù)大劑量組一定時(shí)間內(nèi)的糖耐量明顯高于模型對(duì)照組。
2.2各組胰腺SOD和MDA水平的比較
模型對(duì)照組的SOD水平明顯低于空白對(duì)照組,MDA水平明顯高于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.01或P<0.05)。中藥組分干預(yù)大劑量組的SOD水平明顯高于模型對(duì)照組,MDA水平明顯低于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.01)。中藥組分干預(yù)小劑量組的SOD水平高于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。中藥組分干預(yù)小劑量組的MDA水平明顯低于模型對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P<0.01)(表2)。
2.3各組形態(tài)學(xué)改變
光鏡顯示,相較于空白對(duì)照組,模型對(duì)照組的胰島有明顯萎縮和消失;胰島細(xì)胞廣泛存在空泡變性,局部有玻璃樣變性,周圍有炎性細(xì)胞浸潤,中藥干預(yù)組上述病理改變明顯改善并恢復(fù)(圖1~4)。
3討論
糖尿病作為動(dòng)脈粥樣硬化和高血壓等心血管疾病的獨(dú)立危險(xiǎn)因素,嚴(yán)重影響著人類的健康和生活質(zhì)量。近年來,隨著糖尿病發(fā)病率和死亡率的提升,尋找有效的治療藥物成為糖尿病防治的關(guān)鍵。本實(shí)驗(yàn)采用一次性腹腔注射鏈脲佐菌素的方法破壞胰島建立大鼠糖尿病模型,病理切片顯示模型組相較于空白組胰島明顯變小,胰島細(xì)胞發(fā)生玻璃樣變性并伴有炎癥反應(yīng),胰島有明顯破壞。注射后鏈脲佐菌素1周和10周的模型對(duì)照組的血糖水平顯著高于空白對(duì)照組,藥物干預(yù)組的血糖水平明顯低于模型對(duì)照組,提示三七皂苷、麥冬、黃芪等中藥多糖組分具有明顯的降糖作用;藥物干預(yù)組病理切片顯示胰島B細(xì)胞的病理變化有明顯改善,推測藥物干預(yù)組的降糖作用與改善胰島B細(xì)胞分泌功能密切相關(guān)。口服葡萄糖耐量試驗(yàn)(OGTT)是測量機(jī)體對(duì)葡萄糖的耐受能力,通過一定時(shí)間內(nèi)血糖濃度的變化,推測胰島素的分泌情況及對(duì)血糖的調(diào)節(jié)能力,是糖尿病的確診試驗(yàn)。OGTT常作為判定某種藥物降糖效果的一種評(píng)價(jià)指標(biāo)。有研究顯示血清超敏C-反應(yīng)蛋白(hs-CRP)、同型半胱氨酸(Hcy)、光抑素C(CysC)水平在糖代謝異常早期就已經(jīng)升高,并參與了糖尿病動(dòng)脈硬化的進(jìn)程[9-13]。糖耐量下降會(huì)增加心腦血管系統(tǒng)疾病的發(fā)病率,本實(shí)驗(yàn)顯示藥物干預(yù)組(大劑量組和小劑量組)的糖耐量明顯糾正,這與病理顯示的胰島B細(xì)胞的病理變化明顯改善結(jié)果相一致。SOD具有超氧陰離子自由基的清除能力,其活性代表著機(jī)體抗氧化能力(氧自由基的清除能力);MDA是衡量組織過氧化損傷程度的指標(biāo)。
綜上所述,本實(shí)驗(yàn)對(duì)胰腺SOD水平和MDA水平的檢測結(jié)果顯示復(fù)方中藥組分對(duì)胰腺組織的過氧化物損傷有所糾正,進(jìn)而改善機(jī)體對(duì)血糖的調(diào)節(jié)能力,其在預(yù)防和降低糖尿病心腦血管并發(fā)癥方面具有積極作用[10-16]。
[參考文獻(xiàn)]
[1]雷定超,張?zhí)m蘭,周水平,等.中藥治療糖尿病現(xiàn)狀與研究進(jìn)展[J].中國醫(yī)藥導(dǎo)報(bào),2012,9(21):8-9,12.
[2]宋光熠,姜雅秋,李屹,等.二多糖膠囊對(duì)實(shí)驗(yàn)性糖尿病大鼠血糖的影響[J].吉林醫(yī)學(xué),2007,28(15):1685-1686.
[3]Clarke WL,Cox D,Gonder-Frederick LA,et al.Evaluating clinicalaccuracy of systems for self-monitoring of blood glucose[J].Diabetes Care,1987,10(5):622-628.
[4]Weinstein RL,Schwartz SL,Brazg RL,et al.Accuracy of the 5-day Free Style Navigator continuous glucose monitoring system comparison with frequent laboratory reference measurements[J].Diabetes Care,2007,30(5):1125-1130.
[5]張嫻,劉莉.白藜蘆醇粉對(duì)2型糖尿病患者血糖及超氧化物歧化酶(SOD)、過氧化氫酶(CAT)、谷胱甘肽過氧化物酶(GSH-Px)、丙二醛(MDA)的影響[J].河北中醫(yī),2017,39(1):19-22.
[6]李屹,張建軍.海鞘對(duì)實(shí)驗(yàn)性糖尿病大鼠非酶糖基化的影響[J].遼寧醫(yī)學(xué)院學(xué)報(bào),2008,29(2):125-127.
[7]劉文艷,劉曉蘇.復(fù)方中藥組分抗糖尿病大鼠糖過氧化作用[J].中國老年學(xué)雜志,2013,33(20):5026-5027.
[8]車光升,宋光熠.海鞘對(duì)糖尿病大鼠血漿內(nèi)皮素-1影響[J].中國公共衛(wèi)生,2009,25(5):571-572.
[9]梁峰,胡大一,沈珠軍.2014 美國糖尿病指南:糖尿病診療標(biāo)準(zhǔn)[J].中華臨床醫(yī)師雜志(電子版),2014,8(6):151-159.
[10]安梅,周瑾,陳曉宇.白藜蘆醇藥理學(xué)作用的研究進(jìn)展[J].腫瘤藥學(xué),2014,4(4):242-246.
[11]谷雨明,黃延芹,徐云生.現(xiàn)代中醫(yī)治療糖尿病的辨證體系研究進(jìn)展[J].河北中醫(yī),2015,37(10):1587-1589.
[12]陸菊明.預(yù)混胰島素在糖尿病治療中的應(yīng)用—2010年《中國2型糖尿病防治指南》解讀[J].中華糖尿病雜志,2011,3(2):187-189.
[13]金萍,陳濤,石秀娥,等.外周血指標(biāo)檢測在動(dòng)脈粥樣硬化性腦梗死中的應(yīng)用研究[J].國際檢驗(yàn)醫(yī)學(xué)雜志,2017, 38(11):143-145.
[14]王敏.糖耐量減低與動(dòng)脈粥樣硬化相關(guān)性的研究進(jìn)展[J].醫(yī)學(xué)診斷,2015,5(1):1-5.
[15]陳濤,石秀娥,王一萍,等.糖耐量異常人群血清hs-CRP、Hcy、CysC水平與頸動(dòng)脈粥樣硬化的相關(guān)性研究[J].國際檢驗(yàn)醫(yī)學(xué)雜志,2018,39(15):1838-1841.
[16]劉晶晶,林桂明,王麗穎,等.PDTC通過抑制NF-κB及Wnt/β-catenin信號(hào)通路修復(fù)1型糖尿病大鼠主動(dòng)脈病變[J].中國藥理學(xué)通報(bào),2018,34(11):1622-1627.
(收稿日期:2018-11-26 本文編輯:祁海文)