本文選題：利福平 + 肝損傷�。� 參考：《第三軍醫(yī)大學》2017年碩士論文

【摘要】：研究背景及目的最近幾年,結核病的發(fā)病率較往年有了顯著升高的趨勢,因結核造成的死亡率也有了明顯的提高,居各類傳染病之首。在世界范圍來說,中國是結核病最多的國家之一[1-2]。在抗結核病的治療中,抗結核藥所致的肝損害的發(fā)病率約為2%-28%[3-5],在抗結核藥所致的不良反應中,肝損害占首位,而重度肝損傷的發(fā)病率約為1%-2%[6-7],但引起肝毒性的機制還未明確。因此,我們需要更多的研究,以找到抗結核藥造成肝損害的機制。盡量減少抗結核藥在使用過程中導致的肝損害,或者尋找新的抗結核藥。這也是臨床上一直想解決的問題。利福平(rifampicin,RIF)是屬于利福霉素類的半合成抗生素,它能夠抑制細菌RNA的合成,可用于肺結核、腸結核、結核性腹膜炎、關節(jié)結核等各種結核桿菌感染,以及腸球菌屬等感染的治療。是常用的抗結核藥物之一,在臨床上廣泛運用。因而,它也成了我國藥物引起的急性肝損傷的常見的主要原因。利福平有許多副反應,肝臟毒性是它最為常見的不良反應,長期使用利福平可引起藥物性肝損傷,因而嚴重影響了它在臨床上的使用[8-11]。目前對利福平的肝損傷機制是不清楚的。在抗結核過程中沒有證據(jù)證明利福平在肝臟代謝當中會出現(xiàn)毒性產物[12]。本文通過給予小鼠不同劑量的利福平,觀察利福平是否致小鼠肝損傷,以及給予不同劑量的利福平后小鼠肝損傷的特點,為進一步探討利福平致肝損傷的機制奠定基礎。并構建ABCC2基因過表達細胞株,為今后研究ABCC2基因,MRP2蛋白的表達和定位與利福平導致的膽汁淤積的關系奠定基礎。方法1.利福平致小鼠肝損傷特點的研究方法:將24只健康雌性ICR小鼠用隨機數(shù)字法分成4組,每組各6只,分別為對照組、低、中、高劑量組。每天早上8點半定時灌胃1次,連續(xù)2周。低劑量組每只小鼠按每天100mg/kg給予利福平;中劑量組每只小鼠按每天200mg/kg給予利福平;高劑量組每只小鼠按每天400 mg/kg給予利福平;對照組:予以等容積的羧甲基纖維素鈉的水溶液,以作對照。末次給藥后禁食6 h取材,收集小鼠血液和肝組織樣品進行血清學檢測并觀察小鼠肝臟病理改變。2.ABCC2基因過表達細胞株的構建:(1)ABCC2-eGFP定點整合載體構建,(2)電轉HEK-293細胞,篩選陽性克隆。結果1.各組小鼠間所有肝生化指標無明顯統(tǒng)計學差異。2.各實驗組小鼠肝臟均發(fā)生明顯病理改變。3.肝HE染色示:各實驗組小鼠肝細胞排列紊亂,部分肝細胞腫脹、見空泡狀的細胞,考慮為不同程度的脂肪變性細胞,并且在匯管區(qū)周圍出現(xiàn)大量炎性細胞浸潤,可見散在的膽汁淤積。其中高劑量組表現(xiàn)最為明顯,出現(xiàn)廣泛的炎性細胞浸潤及肝細胞脂肪變性。4.肝電鏡照片示:各實驗組肝臟組織電鏡照片可見肝細胞包漿中充滿大量的脂肪滴,可見部分細胞核萎縮,部分毛細膽管輕微擴張、其內可見膽汁淤積,部分線粒體、內質網(wǎng)腫脹斷裂。也是在高劑量組肝臟病理改變最明顯。5.構建了ABCC2基因過表達細胞株。結論連續(xù)2周給予不同劑量的利福平均可引起小鼠肝損傷,主要表現(xiàn)為肝細胞脂肪變性、炎性細胞浸潤、部分細胞核核萎縮,部分線粒體腫脹,內質網(wǎng)腫脹以及散在的膽汁淤積,利福平對小鼠肝臟的損傷呈濃度依賴性的,但其機制不明確有待進一步研究。成功構建ABCC2基因過表達細胞株。目的這個meta分析的目的是要闡明SLCO1B1 T521C多態(tài)性對他汀類藥物血液濃度的影響。方法通過計算機檢索pubmed、Cochrane Library、EMBASE、中國生物醫(yī)學文獻數(shù)據(jù)庫、相關期刊論文、維普、萬方等數(shù)據(jù)庫,共檢索到的25篇相關研究并提取數(shù)據(jù)。提取AUC0-6h、AUC0-12h、AUC0-24h、AUC0-inf、Cmax和CL/F等藥代動力學參數(shù)。用標準均數(shù)差(SMD)和95%的可信區(qū)間(95%CI)比較野生型(TT)基因型和突變基因型(TC、CC、TC+CC)。亞組分析根據(jù)種族人群、他汀類藥物的類型進行。用敏感性分析對各項研究進行異質性檢驗。從納入研究提取資料然后用rev Man5.3進行Meta分析。結果共納入25篇文獻,包含903例病例,Meta分析結果顯示:全部研究的AUC0-inf標準均數(shù)差TT-TC為0.51(95%CI:0.29,0.72),TT-CC為1.80(95%CI:1.31,2.29),TT-(TC+CC)為0.78(95%CI:0.57,0.99);總體而言,突變基因型的AUC0-inf的標準均數(shù)差較高。在白種人和亞洲人的亞組分析得出同樣的結果。全部研究的Cmax標準均數(shù)差TT-TC為0.48(95%CI:0.18,0.77),TT-CC為1.24(95%CI:0.77,1.70),TT-(TC+CC)為0.65(95%CI:0.43,0.87);突變基因型的Cmax的標準均數(shù)差也較高。在白種人和亞洲人的亞組分析也得出同樣的結果。全部研究的CL/F標準均數(shù)差TT-TC為-1.01(95%CI:-1.91,-0.11),TT-CC為-1.51(95%CI:-2.08,-0.94),TT-(TC+CC)為-1.07(95%CI:-1.55,-0.59),突變基因型的CL/F的標準均數(shù)差較低。結論SLCO1B1 T521C等位基因可能對羥甲基戊二酰輔酶還原酶抑制劑藥代動力學起著重要影響。
[Abstract]:In recent years, the incidence of tuberculosis has been significantly higher than in previous years, and the death rate caused by tuberculosis has also increased significantly. In the world, China is one of the most tuberculosis countries, [1-2]., in the treatment of anti nodule disease, the liver damage caused by anti tuberculosis drugs. The incidence of the disease is about 2%-28%[3-5]. In the adverse reactions caused by anti tuberculous drugs, liver damage is the first, and the incidence of severe liver injury is about 1%-2%[6-7], but the mechanism of hepatotoxicity is not clear. Therefore, we need more research to find the mechanism of anti tuberculosis drug causing liver damage. Rifampicin (rifampicin, RIF) is a semi synthetic antibiotic of rifamycin, which inhibits the synthesis of bacterial RNA and can be used in tuberculosis, intestinal tuberculosis, tuberculosis peritonitis, tuberculosis of joint, and Enterococcus, and Enterococcus. It is one of the commonly used antituberculous drugs and is widely used in clinical practice. Therefore, it has also become a common cause of acute liver injury caused by drugs in our country. Rifampin has many side effects. Liver toxicity is the most common adverse reaction. The long-term use of rifampin can cause drug induced liver damage and therefore serious shadow. The mechanism of the clinical use of [8-11]. is not clear. There is no evidence that rifampin may have a toxic product in the liver metabolism during the anti tuberculosis process, [12]., by giving rifampicin at different doses in mice, to observe whether rifampin causes liver damage in mice and to give different doses. The characteristics of liver injury in Li Fuping's post Li Fuping mice lay the foundation for further study of the mechanism of Li Fuping induced liver injury, and constructed the overexpressed cell line of the ABCC2 gene to lay the foundation for the future study of the relationship between the expression and location of the ABCC2 gene, the expression of MRP2 protein and the cholestasis caused by Li Fuping. Methods: 24 healthy female ICR mice were divided into 4 groups by random number method, 6 rats in each group, respectively, the control group, low, middle, high dose group. Every day at 8:30 every morning, the stomach was given 1 times for 2 weeks. Each mouse in the low dose group was given rifampin per day at 100mg/kg; each mouse in the middle dose group was given rifampin at 200mg/kg per day; the high dose group per mouse was given every day. The mice were given rifampicin at 400 mg/kg per day; the control group was treated with the same volume of sodium carboxymethyl cellulose water as the control. After the last dose of 6 h, the samples of blood and liver tissues of mice were collected for serological examination and the construction of.2.ABCC2 gene overexpressed cell lines in the liver pathological changes of mice was observed: (1) ABCC2-eGFP determination. Point integration carrier construction, (2) electric transfer of HEK-293 cells and screening positive clones. Results there was no significant difference in all liver biochemical indexes between each group of 1. groups. The liver of all mice in each group had obvious pathological changes in.3. liver HE staining: the liver cells in each experimental group were in disorder, some of the hepatocytes were swollen, and the vacuolated cells were considered. A large number of inflammatory cells were infiltrated around the manifold and scattered cholestasis was seen around the pipe area. The high dose group showed the most obvious, extensive inflammatory cell infiltration and hepatocyte fatty degeneration.4. liver electron microscopy. A large number of fat drops, visible part of the nucleus atrophy, partial capillary bile duct dilated slightly, it can be seen in the cholestasis, some mitochondria, the endoplasmic reticulum swelling and fracture. Also in the high dose group, the most obvious liver pathological changes.5. constructed the ABCC2 gene overexpressed cell line. Conclusion the 2 weeks of continuous dose of rifampin can cause the liver of mice. The main manifestations were hepatocyte fatty degeneration, inflammatory cell infiltration, partial nucleus atrophy, partial mitochondria swelling, endoplasmic reticulum swelling and scattered cholestasis. Rifampin was dependent on the damage of liver in mice, but the mechanism was not clear. The ABCC2 gene overexpressed cell line was successfully constructed. The purpose of this meta analysis is to clarify the effect of SLCO1B1 T521C polymorphism on the blood concentration of statins. Methods 25 related studies and extraction data were retrieved by computer retrieval of PubMed, Cochrane Library, EMBASE, Chinese biomedical literature database, Chinese journal full text database, VIP and Wanfang Data base. Pharmacokinetic parameters, such as AUC0-6h, AUC0-12h, AUC0-24h, AUC0-inf, Cmax and CL/F, compared wild type (TT) genotypes and mutant genotypes (TC, CC, TC+CC) with standard mean number difference (SMD) and 95% confidence interval (95%CI). Subgroups analyzed the types of statins based on racial populations. Sensitivity analysis was used to test the heterogeneity of various studies by sensitivity analysis. A total of 25 articles were included in 25 articles and 903 cases were included. The results of Meta analysis showed that the average number difference TT-TC of the AUC0-inf standard was 0.51 (95%CI:0.29,0.72), TT-CC was 1.80 (95%CI:1.31,2.29), TT- (TC+CC) was 0.78 (95%CI:0.57,0.99); in general, the mutant genotype The standard mean difference of 0-inf was high. The same results were obtained in the subgroup of white people and Asians. The average number difference TT-TC of the Cmax standard was 0.48 (95%CI:0.18,0.77), TT-CC was 1.24 (95%CI:0.77,1.70), TT- (TC+CC) was 0.65 (95%CI:0.43,0.87); the standard mean mean difference of the mutant genotype was also higher. The subgroup analysis also obtained the same results. The CL/F standard mean difference TT-TC of all studies is -1.01 (95%CI:-1.91, -0.11), TT-CC is -1.51 (95%CI:-2.08, -0.94), TT- (TC+CC) is -1.07. Pharmacokinetics of preparation plays an important role.

【學位授予單位】：第三軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R52;R575

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本文編號：1885371