雙環(huán)醇保肝抗炎作用研究進(jìn)展

1、百賽諾（雙環(huán)醇片）保肝抗炎作用研究進(jìn)展,肝細(xì)胞損傷和肝臟炎癥壞死,肝細(xì)胞損傷是各型肝病共同的病理基礎(chǔ)及共同表現(xiàn); 導(dǎo)致肝細(xì)胞變性、凋亡、壞死最終導(dǎo)致肝衰竭。肝臟炎癥壞死及其所致的肝纖維化是疾病進(jìn)展的主要病理學(xué)基礎(chǔ)。在對(duì)病因治療的基礎(chǔ)上有效控制肝組織炎癥，有可能減少肝細(xì)胞破壞和延緩肝纖維化的發(fā)展。,轉(zhuǎn)氨酶的功能及臨床意義,血清氨基轉(zhuǎn)移酶以肝臟含量最為豐富,臨床中用于血清學(xué)診斷主要為： 谷丙轉(zhuǎn)氨

2、酶 (ALT):肝>腎>心>肌肉 谷草轉(zhuǎn)氨酶 (AST):心>肝>肌肉>腎在肝內(nèi)，ALT全部存在于肝細(xì)胞漿中；AST 80%存在于線粒體內(nèi)，20%在胞漿內(nèi)。 當(dāng)肝細(xì)胞發(fā)生炎癥、壞死、中毒等造成肝細(xì)胞受損時(shí)，轉(zhuǎn)氨酶會(huì)釋放入血液，血清轉(zhuǎn)氨酶升高。ALT水平可以比較敏感地監(jiān)測(cè)到肝臟是否受到損害。當(dāng)肝細(xì)胞嚴(yán)重?fù)p傷波及線粒體時(shí)AST也會(huì)進(jìn)入血中。,肝損傷的基本治療策略,病因治療：

3、 消除各種致肝損害的原因?qū)ΠY治療： 降酶、退黃、消除其他癥狀保護(hù)肝功能：保護(hù)肝細(xì)胞、消除炎癥損害替代肝功能：促進(jìn)肝細(xì)胞生長(zhǎng)、協(xié)助解毒功能 的藥物、人工肝替代療法綜合治療： 上述療法＋營(yíng)養(yǎng)支持肝臟移植： 原位肝移植、活體肝移植,,,病毒性肝炎,酒精性肝病,藥物性肝病,肝纖維化,肝硬化,炎癥反應(yīng),肝癌肝功能衰竭,纖維組織增生，星狀細(xì)胞活化,

4、對(duì)因治療,,肝細(xì)胞膜損傷,非酒精性 脂肪肝,脂質(zhì)代謝紊亂,能量代謝紊亂,,,肝細(xì)胞壞死,自由基損傷,肝病發(fā)展不同階段的治療重點(diǎn),,,保肝藥物作用環(huán)節(jié),,對(duì)癥治療,抗纖維化抗硬化,抗癌治療肝移植,抗病毒,,其他原因,,,臨床常用的抗炎保肝藥物,雙環(huán)醇甘草酸制劑水飛薊素類還原型谷胱苷肽多烯磷脂酰膽堿硫普羅寧等,-不是肝臟和血清ALT和AST活性的抑制劑-不是肝臟ALT酶蛋白合成的抑制劑,7,百賽諾對(duì)轉(zhuǎn)氨酶的作用,百賽

5、諾不是肝臟和血清ALT和AST活性的抑制劑 大鼠肝臟、血清直接溫孵法，發(fā)現(xiàn)ALT活性不降低,百賽諾不抑制小鼠肝臟和血清ALT、AST活性,Geng-Tao Liu, Yan Li, et al. Mechanism of protective action of bicyclol against CCl4-induced liver injury in mice. Liver International, 200

6、5, 25(4):872-879 .,9,小鼠肝臟ALT酶蛋白純化,免疫家兔,兔抗小鼠肝臟ALT抗體制備,抗體+小鼠給藥后肝勻漿進(jìn)行免疫火箭電泳測(cè)定肝臟ALT蛋白水平,1-4: 正常組 82.3±6.85-9: 雙環(huán)醇組 82.0±7.4 Protein: 170mg/each,正常和給藥小鼠肝勻漿ALT免疫火箭電泳測(cè)定,百賽諾不影響肝臟ALT酶蛋白含量,Geng-Tao Liu, Y

7、an Li, et al. Mechanism of protective action of bicyclol against CCl4-induced liver injury in mice. Liver International, 2005, 25(4):872-879 .,肝細(xì)胞保護(hù)劑對(duì)轉(zhuǎn)氨酶的作用（體內(nèi)）,臨床耐受性試驗(yàn),志愿者：6.25-150mg, tidx4周對(duì)志愿者血清ALT，AST活性無(wú)影響。,（結(jié)果已發(fā)表在

8、國(guó)內(nèi)外核心刊物）,清除自由基、抗氧化,Liu GT, Li Y, Wei HL, et al. Mechanism of protective action of bicyclol against CCl4 induced liver injury in mice. Liver International. 2005, 25(4):872-879 .,Effect of bicyclol on the levels of CCl3 r

9、adical as detected by ESR in liver microsomes,Bicyclol (200, 300 mg/kg) was given orally to mice three times before alcohol treatment. Mice were sacrificed at 12 h and 6 h after alcohol administration for GSH content d

10、etermination respectively. Data were expressed as means±SD (n=8).*, P<0.05, ***, P<0.001 vs. control group; #, P<0.05 vs. alcohol group.,Zhao J, Chen H, Li Y. Eur J Pharmacol. 2008 ;586(1-3):322-331.,Fig.

11、10. Time-course changes in plasma endotoxin level in acute alcohol intoxicated mice. Alcohol (6 g/kg) was administered to mice by gavage. The animals were sacrificed at 1.5, 3, 6, and 12 h after alcohol administration. D

12、ata were expressed as means±SD (n=6). ??, P<0.01 vs. control group.,Fig.11. Effect of bicyclol on plasma endotoxin level in acute alcohol-intoxicated mice. Bicyclol (200, 300 mg/kg) was given orally to mice three

13、 times before alcohol treatment. Mice were sacrificed at 1.5 h after alcohol administration.Data were expressed as means±SD (n=6). ??, P<0.01 vs. control group;##, P<0.01 vs. alcohol group.,Effect of bicyclol

14、on plasma endotoxin level in acute alcohol-intoxicated mice,Zhao J, Chen H, Li Y. Eur J Pharmacol. 2008 ;586(1-3):322-331.,抗肝損傷-對(duì)肝細(xì)胞/線粒體膜形態(tài)的保護(hù)作用,藥物對(duì)肝線粒體膜的保護(hù)作用（體內(nèi)）A，B-正常對(duì)照，C，D-肝損傷，E，F(xiàn)-給藥后,藥物對(duì)肝細(xì)胞膜的保護(hù)作用（體外）,Hui-Ping Wang,

15、Yan Li. Protective effect of bicyclol on acute hepatic failure induced by lipopolysaccharide and D-galactosamine in mice. European Journal of Pharmacology. 2006, 534(1-3):194-201.,[1]趙冬梅，劉耕陶.雙環(huán)醇對(duì)對(duì)乙酰氨基酚致小鼠肝線粒體損傷的保護(hù)作用.中國(guó)新藥

16、雜志，2002，7（11）：536-540[2]李燁，李燕，劉耕陶.雙環(huán)醇對(duì)實(shí)驗(yàn)性肝纖維化的防護(hù)作用及分子機(jī)制.中華醫(yī)學(xué)雜志,2004,84(24):2096-2101[3]李 燁，戴國(guó)煒，李 燕，劉耕陶.雙環(huán)醇對(duì)撲熱息痛弓l起小鼠肝臟能量代謝和線粒體功能障礙的影響.藥學(xué)學(xué)報(bào).2001，36（10）：723-726,抗肝損傷-對(duì)線粒體功能的保護(hù)作用,線粒體ATP酶活性,線粒體腫脹度,線粒體膜流動(dòng)性,抗肝損傷-對(duì)肝臟病理形態(tài)的保護(hù)作用

17、,,[1]Liu GT, Li Y, Wei HL, et al. Mechanism of protective action of bicyclol against CCl4 induced liver injury in mice. Liver International. 2005, 25(4):872-879 .[2]Geng Tao Liu. Bicyclol: A Novel Drug For Treating Chro

18、nic Viral Hepatitis B and C.Medicinal Chemistry,2009,5,29-43.[3]莫成林, 李燁, 李燕. 雙環(huán)醇對(duì)小鼠慢性酒精性肝損傷的保護(hù)作用[ J ]. 中華醫(yī)學(xué)雜志, 2005, 85 (48) : 3409-3413.,Fig. 12. Localization of liver TNF-α and CD14 expression in acute alcohol-intoxic

19、ated mice. Bicyclol (200, 300 mg/kg) was given orally to mice (n=5) three times before alcohol treatment. Mice were sacrificed at 12 h after alcohol administration. 1: expression of TNF-α; 2: expression of CD14. a: Cont

20、rol; b: Alcohol;c: Pretreatment with Bicyclol. Arrows: Positive cells. Original magnification×100.,抗肝損傷分子機(jī)制-抑制炎癥因子表達(dá),Fig. 9. Effects of bicyclol on hepatic TNF-α and IL-1βmRNA expression in acute alcohol-intoxicat

21、ed mice. Bicyclol (200, 300 mg/kg) was given orally to mice three times before alcohol treatment. Mice were sacrificed at 12 h after alcohol administration. (A): lane 1–2, Control; lane 3–4, Alcohol; lane 5–6, By 200 mg/

22、kg; lane 7–8, By 300 mg/kg. (B): Ratio of PCR products relative to GAPDH. Data were expressed as means±SD (n=4). *, Pb0.05 vs. control group; #, Pb0.05, ###, Pb0.001 vs. alcohol group.,[1]Zhao J, Chen H, Li Y. Prot

23、ective effect of bicyclol on acute alcohol- induced liver injury in mice [ J ]. Eur J Pharmacol, 2008, 586 (123) :322-331.[2]李燁，李燕，劉耕陶.雙環(huán)醇對(duì)實(shí)驗(yàn)性肝纖維化的防護(hù)作用及分子機(jī)制.中華醫(yī)學(xué)雜志,2004,84(24):2096-2101,抗肝損傷分子機(jī)制-抑制炎癥導(dǎo)致的肝細(xì)胞凋亡,趙冬梅、劉耕陶.

24、雙環(huán)醇對(duì)刀豆蛋白A所致小鼠肝細(xì)胞核DNA損傷的保護(hù)作用. 中華醫(yī)學(xué)雜志，2001, 81(14):844-848 .,A: 100bp.DNA條帶標(biāo)準(zhǔn)品B C: 正常對(duì)照組D E F: ConA模型對(duì)照組G H I: 百賽諾150mg/kg,Wang H, Li Y. Eur J Pharmacol. 2006 ;534(1-3):194-201.,Effect of bicyclol on liver injury in

25、duced by lipopolysaccharide/D-galactosamine in mice,Liver specimens were obtained at 6 h after LPS/Dgalactosamine injection. (A) Normal control; (B) (C) carboxymethyl cellulose vehicle administration 1 h before LPS/GalN

26、injection; (D) bicyclol 300 mg/kg administration for 3 times 1 h before LPS/D-galactosamine injection; (E) bicyclol 300 mg/kg administration once 1 h before LPS/D-galactosamine injection. Original magnification X100.,小結(jié)

27、,雙環(huán)醇體外對(duì)血清ALT、AST的活性無(wú)直接抑制作用，體內(nèi)給藥對(duì)肝臟ALT蛋白水平無(wú)影響。臨床志愿者口服藥物對(duì)轉(zhuǎn)氨酶活性也無(wú)抑制作用。 保肝藥的降酶作用來(lái)自--肝細(xì)胞膜和線粒體膜形態(tài)和功能的改善-抑制炎癥因子及相關(guān)受體的表達(dá)-抑制炎癥導(dǎo)致的肝細(xì)胞凋亡,19,雙環(huán)醇的臨床使用依據(jù),《慢性乙型肝炎防治指南(2010年更新版)》《非酒精性脂肪性肝病診療指南(2010年修訂版)》《酒精性肝病診療指南(2010年修訂版)》《

28、河南省新農(nóng)合按病種付費(fèi)臨床路徑“酒精性肝炎”》 （豫衛(wèi)醫(yī)改（2012）4號(hào)文件）,1.雙環(huán)醇治療酒精性肝病,醫(yī)院：衛(wèi)生部中日友好醫(yī)院負(fù)責(zé)人：馬安林試驗(yàn)組： 54例，給予百賽諾50mg，tid. 對(duì)照組： 49例，給予多烯磷脂酰膽堿456mg，tid. 2組均連續(xù)用藥36周，試驗(yàn)組23例、對(duì)照組21例患者完成治療前后2次肝穿刺活組織檢查,馬安林, 郭新珍, 劉霞, 等.雙環(huán)醇

29、與多烯磷脂酰膽堿治療酒精性肝病的療效比較. 中華肝臟病雜志. 2011, 19(6):471-472,雙環(huán)醇與多烯磷脂酰膽堿治療酒精性肝病的療效比較,馬安林，郭新珍，劉霞.中華肝臟病雜志.2011.l9(6):471-472.,雙環(huán)醇與多烯磷脂酰膽堿治療酒精性肝病的療效比較,馬安林，郭新珍，劉霞.中華肝臟病雜志.2011.l9(6):471-472.,雙環(huán)醇與多烯磷脂酰膽堿治療酒精性肝病的療效比較,馬安林，郭新珍，劉霞.中華肝臟病雜志

30、.2011.l9(6):471-472.,雙環(huán)醇與多烯磷脂酰膽堿治療酒精性肝病的療效比較,馬安林，郭新珍，劉霞.中華肝臟病雜志.2011.l9(6):471-472.,研究結(jié)果及結(jié)論,百賽諾治療組ALT及AST的下降速度和程度優(yōu)于多烯磷脂酰膽堿對(duì)照組。 百賽諾治療及對(duì)照組36周后可使血清GST—PX水平顯著上升，MDA水平顯著下降。 通過(guò)比較治療前后的超聲影像學(xué)表現(xiàn)，我們看到兩組治療前后均有一定程度的改善，而且百賽諾可在

31、一定程度上改善肝內(nèi)脂肪沉積、炎癥死及纖維化，尤其對(duì)于炎癥的改善程度。,2.百賽諾治療非酒精性脂肪肝,注: 1、患者診斷標(biāo)準(zhǔn)符合2006年2月《非酒精性脂肪性肝病診療指南》 2、隨機(jī)分組采用 SPSS15.0統(tǒng)計(jì)軟件 3、治療期間均未使用其他保肝藥物,78例20歲～64歲非酒精性脂肪性肝病患者，隨機(jī)分為兩組采用減輕體重(<1200g/周)為前提的基礎(chǔ)治療聯(lián)合藥物治療，兩組療程均為24周,治療前后檢測(cè)：

32、人體學(xué)指標(biāo)、B超、肝功能檢測(cè)、肝臟組織學(xué)檢查,蘇紅領(lǐng),韓英,樊代明,等.雙環(huán)醇與多烯磷脂酰膽堿治療非酒精性脂肪肝的療效比較.中華肝臟病雜志.2011,19(7):552-553.,10,5,17,11,32,23,20,30,,,25,15,0,35,%,,,研究結(jié)果-百賽諾組部分應(yīng)答率優(yōu)于對(duì)照組,應(yīng)答標(biāo)準(zhǔn)：(1)完全應(yīng)答：ALT復(fù)常；超聲遠(yuǎn)場(chǎng)回聲衰減程度較治療前改善，且GST-PX和MDA至少l項(xiàng)較治療前改善；脂變、炎癥及壞死積分

33、較治療前減少2分以上。(2)部分應(yīng)答：ALT復(fù)常；超聲檢查指標(biāo)改善不顯著；組織學(xué)改變不明顯。(3)無(wú)應(yīng)答：未達(dá)到上述指標(biāo)者,1.百賽諾顯著改善肝功能指標(biāo),研究結(jié)果—百賽諾顯著改善肝功和血脂指標(biāo),ALP：堿性磷酸酶 GGT：谷氨酰轉(zhuǎn)肽酶,2.百賽諾明顯改善血脂指標(biāo),3.百賽諾明顯改善B超積分,研究結(jié)果—百賽諾顯著改善B超積分和炎癥,近場(chǎng)回聲增高、灶性高回聲或肝光點(diǎn)增粗各計(jì)1分；遠(yuǎn)場(chǎng)回聲衰減、肝腫大、肝內(nèi)管道系統(tǒng)顯示不清或無(wú)

34、法辨認(rèn)各計(jì)2分。,4.百賽諾改善脂肪變性、炎癥、纖維化,*與對(duì)照組相比，P<0.05,百賽諾治療后脂肪變、炎癥、纖維化不同程度減輕,,蘇木精一伊紅染色,脂肪變性和炎癥壞死,治療前,治療后,纖維化表現(xiàn),網(wǎng)狀纖維染色,檢查項(xiàng)目：ALT、AST、TBiL,3.百賽諾防治化療藥物所致肝損害,周建鳳, 陳書(shū)長(zhǎng), 白春梅, 等. 雙環(huán)醇片防治化療藥物性肝損害的研究. 肝臟, 2007, 12(4):286-287 .,結(jié)果-百賽諾對(duì)肝損害的治

35、療作用,結(jié)果-百賽諾對(duì)肝損害的預(yù)防作用,本研究表明口服雙環(huán)醇片可有效治療化療藥物性肝 損害，中位治療12天后肝功能即顯著恢復(fù)。 化療同時(shí)并用雙環(huán)醇片，患者肝功能損害的發(fā)生率 大為下降，程度也明顯減輕，保障了化療按時(shí)足量 進(jìn)行。,研究結(jié)論,,,Xie W, Shi G, Zhang H, et al. Hepatology International. 2012, 6(2):441-448,A randomize

36、d, multi-central, controlled study of patients with hepatitis B e antigen-positive chronic hepatitis B treated by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,A randomized, multi-central, controlled study of

37、patients with hepatitis B e antigen-positive chronic hepatitis B treated by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,Xie W, Shi G, Zhang H, et al. Hepatology International. 2012, 6(2):441-448,A randomized

38、, multi-central, controlled study of patients with hepatitis B e antigen-positive chronic hepatitis B treated by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,Xie W, Shi G, Zhang H, et al. Hepatology Internati

39、onal. 2012, 6(2):441-448,Xie W, Shi G, Zhang H, et al. Hepatology International. 2012, 6(2):441-448,A randomized, multi-central, controlled study of patients with hepatitis B e antigen-positive chronic hepatitis B treat

40、ed by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,,Fig. 6 Necroinflammation and fibrosis scores of patient No. 2were significantly improvedafter 48 weeks therapy (b) compared with baseline(a) using ADV plus

41、 bicyclol,Fig. 5 Necroinflammation and fibrosis scores of patient No. 1were significantly improved after 48 weeks therapy(b) compared with baseline(a) using ADV plus bicyclol,A randomized, multi-central, controlled stud

42、y of patients with hepatitis B e antigen-positive chronic hepatitis B treated by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,Xie W, Shi G, Zhang H, et al. Hepatology International. 2012, 6(2):441-448,A rando

43、mized, multi-central, controlled study of patients with hepatitis B e antigen-positive chronic hepatitis B treated by adefovir dipivoxil or adefovir dipivoxil plus bicyclol,Xie W, Shi G, Zhang H, et al. Hepatology Inte