本文關(guān)鍵詞： 護(hù)肝片 CCL_4 ALT Ca~(2+) 線粒體　出處：《黑龍江中醫(yī)藥大學(xué)》2012年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的：研究護(hù)肝片降低CCL4肝損傷模型大鼠血清丙氨酸氨基轉(zhuǎn)移酶活性的作用機(jī)制 方法：將Wistar大鼠適應(yīng)性飼養(yǎng)7天后,隨機(jī)分為空白對(duì)照組、模型對(duì)照組、護(hù)肝片治療組,每組.8只。各組大鼠正常飲食,光照時(shí)間7:00am-7:00pm�？瞻讓�(duì)照組和模型對(duì)照組灌胃給予適宜體積的生理鹽水；護(hù)肝片治療組按1700mg/kg灌胃給予護(hù)肝片混懸液,1次/天,連續(xù)9天。末次給藥后1h,模型組及藥物治療組予大鼠腹腔注射20%CCl4橄欖油溶液(0.2ml/100g)；空白對(duì)照組腹腔注射等體積的生理鹽水。造模24h后采集觀察指標(biāo)。比色法測(cè)定血清及肝組織ALT活性；Western blotting測(cè)定肝組織ALT-1、ALT-2相對(duì)含量；RT-PCR測(cè)定肝細(xì)胞ALT mRNA表達(dá)情況；紫外-可見(jiàn)分光光度法測(cè)定線粒體腫脹敏感性及線粒體Na+-K+ATPase、Ca2+—Mg2+ATPase活性熒光分光光度法測(cè)定肝細(xì)胞線粒體膜電位、肝細(xì)胞內(nèi)及肝細(xì)胞內(nèi)游離鈣離子濃度,酶聯(lián)免疫法測(cè)定線粒體滲透性轉(zhuǎn)換孔(MPTP)開(kāi)放程度。 結(jié)果： 1.護(hù)肝片對(duì)CCL4肝損傷模型大鼠ALT活性及含量的影響：與空白組比較,模型組大鼠血清及肝組織ALT活性顯著升高(P0.01)；與模型組比較,護(hù)肝片治療組大鼠血清及肝組織ALT活性水平顯著降低(P0.01)。與空白組比較,模型組肝臟ALT-1含量升高0.47倍,ALT-2升高0.83倍.藥物組ALT-1含量升高0.42倍,ALT-2升高0.54倍。結(jié)合肝組織ALT活性測(cè)定結(jié)果(與空白組比較,模型組肝ALT活性升高1.72倍,藥物組升高1.43倍)可以看出,盡管ALT-1、ALT-2含量變化幅度要小于活性的,但兩者變化趨勢(shì)一致。 2.護(hù)肝片對(duì)肝組織ALT-1、ALT-2mRNA表達(dá)的影響研究結(jié)果：模型組肝組織ALT-1mRNA表達(dá)通量為空白組的0.42；藥物組為0.52。藥物組ALT-2mRNA表達(dá)通量為空白組的0.48；藥物組為0.47。 3.護(hù)肝片對(duì)CCL4肝損傷模型大鼠肝細(xì)胞線粒體功能及細(xì)胞內(nèi)游離鈣離子濃度的影響研究結(jié)果：與空白組比較,模型組大鼠肝細(xì)胞內(nèi)游離鈣離子濃度、肝組織MPTP開(kāi)放程度顯著增加,膜電位、ATPase活性及對(duì)鈣離子誘導(dǎo)腫脹敏感性的均顯著降低；與模型組比較,藥物組大鼠肝細(xì)胞內(nèi)游離鈣離子濃度及肝組織MPTP開(kāi)放程度顯著降低；膜電位、ATPase活性及對(duì)鈣離子誘導(dǎo)腫脹敏感性的均顯著升高。 4.護(hù)肝片對(duì)CCL4肝損傷模型大鼠血清ALT代謝的影響研究結(jié)果：空白組大鼠血清在0～84h內(nèi)呈輕微上升趨勢(shì),但仍處于正常范圍內(nèi)；模型組及模型組大鼠血清在0～84h內(nèi)呈下降趨勢(shì),但二者下降趨勢(shì)相近。藥物組ALT活性下降一半時(shí)間(t1/2)位于72～84h之間,而模型組ALT活性尚未降低至50%。說(shuō)明藥物組ALT活性下降速率高于模型組 結(jié)論： 1、護(hù)肝片具有降低CCL4肝損傷模型血清及肝組織ALT活性的作用；ALT-1、ALT-2含量升高是肝組織ALT活性升高的物質(zhì)基礎(chǔ)；護(hù)肝片可減少損傷肝組織ALT-1、ALT-2含量的升高。 2、CCL4肝損傷模型大鼠肝組織ALT-1、ALT-2mRNA表達(dá)降低,護(hù)肝片使損傷肝組織的ALT-1mRNA表達(dá)有所恢復(fù)。 3、初步確定護(hù)肝片降低血清ALT活性的機(jī)制為保護(hù)線粒體功能,進(jìn)而減少損傷肝細(xì)胞細(xì)胞內(nèi)鈣超載現(xiàn)象,保護(hù)肝細(xì)胞。 4、護(hù)肝片能促進(jìn)CCL。肝損傷模型大鼠血清中ALT的代謝。
[Abstract]:Objective: To study the mechanism of liver protection tablets to reduce the activity of serum alanine aminotransferase in rat model of CCL4 liver injury
Methods: Wistar rats were fed for 7 days, were randomly divided into control group, model control group, Hugan tablets in the treatment group, each group of.8 rats. The rats with normal diet, light time 7:00am-7:00pm. blank control group and model control group were lavaged with appropriate volume of saline; huganpian treatment group by 1700mg/kg perfusion Administration of Hugan tablets suspension, 1 times / day for 9 consecutive days. After the last administration of 1H, the model group and the treatment group was injected intraperitoneally to rats 20%CCl4 olive oil solution (0.2ml/100g); the control group were injected with saline. After 24h model acquisition observed. The determination of ALT the activity of serum and liver tissue Western blotting colorimetric method; Determination of liver tissue ALT-1, ALT-2 relative content; Determination of RT-PCR liver cells ALT mRNA expression; UV Vis spectrophotometric method for the determination of the sensitivity of mitochondrial swelling and mitochondrial Na+-K+ATPase, Ca2+ Mg2+ATPase activity spectrofluorometry was used to measure the mitochondrial membrane potential of liver cells, the concentration of free calcium in hepatocytes and hepatocytes, and the permeability of mitochondrial permeability transition pore (MPTP) was measured by enzyme-linked immunosorbent assay (ELISA).
Result錛，

本文編號(hào)：1555221