参龟保肝升白丸对大鼠实验性肝纤维化的影响及其机制研究
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摘要
吉林省知名老中医经过多年临床实践,总结出由人参、龟板胶、赤芍、首乌、生地黄等十六味中药组成的中药复方制剂——制肝生白颗粒。具有活血化瘀,软坚散结,补养正气的功能。经临床证实,对肝硬化腹水伴白蛋白低下患者具有良好的治疗作用。前期药理研究证明,制肝生白颗粒具有保护肝脏、升高血清白蛋白含量和逆转肝纤维化向肝硬化发展的作用。
由于复方制剂成分的复杂性,有效成分的不确定性以及产品质量的不可控性,导致药品疗效的稳定性减弱,生产成本增加,在某些方面阻碍了中药的广泛应用和现代化发展进程。为提高中药复方制剂质量的可控性,更有效的发挥其治疗功用,减少复杂药味的相互影响,开发中药小复方制剂是中药研究的发展方向。因此,我们将十六味中药进行提炼组成以长白山特产名贵中药人参为君药,辅以龟板胶等四味中药的小复方制剂——参龟保肝升白丸。通过皮下注射CCl4花生油溶液建立大鼠实验性肝纤维化模型,观察参龟保肝升白丸对实验性肝纤维化大鼠血清蛋白、肝功能、肝组织胶原及氧化损伤等指标的影响,并与制肝生白颗粒进行比较,为开发治疗肝纤维化、肝硬化伴白蛋白低下的中药六类新药提供药效依据。
实验结果表明:参龟保肝升白丸4、8g/kg连续给药1个月后,能明显升高实验性肝纤维化大鼠血清白蛋白(ALB)含量,降低血清球蛋白(GLO)含量,显著升高白蛋白/球蛋白(A/G)比值;能明显降低血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)及血清胆红素(BIL)含量,升高胆碱酯酶(CHE)活力;能够显著减少肝组织羟脯氨酸(Hyp)的生成,并明显降低肝组织丙二醛(MDA)含量,升高超氧化物歧化酶(SOD)及谷胱甘肽-S转移酶(GSH-ST)活性。
参龟保肝升白丸虽然减少了药味,其对抗CCl4诱导的大鼠实验性肝纤维化作用与制肝生白颗粒没有显著性差异,但升高白蛋白的效果明显增强,提示新组成的中药小复方制剂具有开发和利用价值。
Liver is one of the most significant organs, with the largest volume and complicated functions. And what is more, it is also be the center which is in charge of participating in metabolism of carbohydrate, lipids and protein. In addition, it has many functions, such as detoxication, hematopoiesis, blood clotting and immunologic barrier. As a digestive gland, it leads excretion of bile, as well as synthesis of many bioactive materials. In Chinese Medicine, liver has the duty of body actions and virus defense, all of these above have been powerfully proved that the healthy of liver is vital to human bodies.
Liver diseases are common in China and are liable to be taken place, especially viral hepatitis. Parts of its patients will change into hepatic cirrhosis, even to hepatic cancer, which bring mass of disasters to them. Pathological changes of liver also can be caused by virus, parasitic infection, diseases poisoning and many other factors, which accrete connective tissue in liver. Thus, it is urgent to do research in pathogenesis and medicine therapies of liver disease all around the world.
Hepatic fibrosis is a character of prophase cirrhosis, and the common pathology reform of chronic hepatitis to cirrhosis. It performs unbalance of extra-cellular matrix (ECM) synthesis and decomposition, which leads to deposit of ECM, and to cause fibrous connective tissue sediment abnormally in liver. Hepatic fibrosis, as an initiative and convertible procedure of hepatic injury, its pathology features are as follows, large amount of collagen fibers I and III forms in headers and lobules of liver, which leads fibers tuberosity and destroy the normal structure, which eventually leads to cirrhosis, degeneration of liver, and sometimes to be cancer. If the disease has existed without control for a long time, it will be aggravated and hepatic lobule and vessels will be rebuilt. Distances, form to pseudolobule, will appear during the sections of veins and headers, and then become to irreversible cirrhosis. In one word, hampering and converting hepatic fibrosis is one of important strategies to cure each kind of chronic hepatitis.
At present, there is lack of efficient anti-fibrosis medicines without obvious side effects, and further more, majority medicines always affect on single joint or with one target, this phenomenon impede the entire process of therapy. On the other hand, Chinese Medicine Recipes have many virtues as little cost, high security and different target points. As a result, lots of patients turn to Chinese Medicine Recipes.
The Chinese Medicine Recipe ZGG using in our research is devised by a famous Chinese physician with high reputation in Jilin Province, which is composed of Panax Ginseng C.A.Meyer, Turtles Gelatin, Radix Paeoniae Rubra Polygonummultiflorum Thunb, and Rehmannia glutinosa as basis. And what is more, it has the function of activating blood circulation, soften lump, loosen knot and tonifying energy. ZGG has already manifested good clinical effects on liver protection in cirrhosis with lower albumin. The former pharmacological research also proved that ZGG has effects of protecting liver, promoting albumin and reversing the cirrhosis forming.
With the characters of Chinese Medicine Recipes, such as unavoidable complication and uncertain active parts of components as well as many difficulties to control its quality, are resulted in the cut of medical effects and high cost of manufacture, all of these hamper the widely use and modern developments of Chinese Medicine Recipes. In order to control the qualities and quantities of Chinese Medicine Recipes, it is necessary to minimize the mutual effects of components in recipes, without therapeutic influences. It is the tendency of research on less components recipes.
Thus, we extract the main basis of ZGG–Panax Ginseng adds Turtles Gelatin to SGP (less components recipe).Through the experimental hepatic fibrosis rats established by hypodermic injection CCl4 and arachis oil, we intend to observe the index of protein level, hepatic function, oxidative damage and tissue collagen, compared with ZGG, in order to provide effective and feasible pharmacodynamic evidences in curing hepatic fibrosis and low albumin diseases with Chinese Medicine Recipes.
The results are as follows. After introgastric-infused for one month, SGP 4、8g/kg can enhance body constitution , reduce liver coefficient, and notable increase albumin (ALB) , A/G and lower globulin (GLO). Furthermore, it markedly decrease the level of aspartate aminotransferase (AST), alanine aminotransferase (ALT), bilirubin (BIL), and increase cholinesterase (CHE) of serum; Moreover, it remarkably downsize the content of malonaldehyde (MDA)and upsize the activity of superoxide dismutase (SOD) and glutathione-S transferase (GSH-ST) in liver .At the same time, it also leads to the drop of hydroxyproline (Hyp) in liver. As the results above, we can demonstrate that SGP has the same effects in enhancing and improving liver situations to ZGG. Thus, SGP may probably affect procedures of synthesis of protein in liver, especially synthesis of albumin, adjustment of olig-albuminemia and inversion of A/G, as well as prevention of collagen abnormally sediment. On the other hand, it maybe also through the mechanism of reducing lipid oxidation, cleaning free radicals and reducing necrosis to resist hepatic fibrosis.
In the end, we can draw a conclusion, despite of less components of SGP, there are identifiable effects to ZGG in conquering hepatic fibrosis. In addition, SGP can striking raise ALB of serum, which means SGP has the promising potential to apply and develop.
由于复方制剂成分的复杂性,有效成分的不确定性以及产品质量的不可控性,导致药品疗效的稳定性减弱,生产成本增加,在某些方面阻碍了中药的广泛应用和现代化发展进程。为提高中药复方制剂质量的可控性,更有效的发挥其治疗功用,减少复杂药味的相互影响,开发中药小复方制剂是中药研究的发展方向。因此,我们将十六味中药进行提炼组成以长白山特产名贵中药人参为君药,辅以龟板胶等四味中药的小复方制剂——参龟保肝升白丸。通过皮下注射CCl4花生油溶液建立大鼠实验性肝纤维化模型,观察参龟保肝升白丸对实验性肝纤维化大鼠血清蛋白、肝功能、肝组织胶原及氧化损伤等指标的影响,并与制肝生白颗粒进行比较,为开发治疗肝纤维化、肝硬化伴白蛋白低下的中药六类新药提供药效依据。
实验结果表明:参龟保肝升白丸4、8g/kg连续给药1个月后,能明显升高实验性肝纤维化大鼠血清白蛋白(ALB)含量,降低血清球蛋白(GLO)含量,显著升高白蛋白/球蛋白(A/G)比值;能明显降低血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)及血清胆红素(BIL)含量,升高胆碱酯酶(CHE)活力;能够显著减少肝组织羟脯氨酸(Hyp)的生成,并明显降低肝组织丙二醛(MDA)含量,升高超氧化物歧化酶(SOD)及谷胱甘肽-S转移酶(GSH-ST)活性。
参龟保肝升白丸虽然减少了药味,其对抗CCl4诱导的大鼠实验性肝纤维化作用与制肝生白颗粒没有显著性差异,但升高白蛋白的效果明显增强,提示新组成的中药小复方制剂具有开发和利用价值。
Liver is one of the most significant organs, with the largest volume and complicated functions. And what is more, it is also be the center which is in charge of participating in metabolism of carbohydrate, lipids and protein. In addition, it has many functions, such as detoxication, hematopoiesis, blood clotting and immunologic barrier. As a digestive gland, it leads excretion of bile, as well as synthesis of many bioactive materials. In Chinese Medicine, liver has the duty of body actions and virus defense, all of these above have been powerfully proved that the healthy of liver is vital to human bodies.
Liver diseases are common in China and are liable to be taken place, especially viral hepatitis. Parts of its patients will change into hepatic cirrhosis, even to hepatic cancer, which bring mass of disasters to them. Pathological changes of liver also can be caused by virus, parasitic infection, diseases poisoning and many other factors, which accrete connective tissue in liver. Thus, it is urgent to do research in pathogenesis and medicine therapies of liver disease all around the world.
Hepatic fibrosis is a character of prophase cirrhosis, and the common pathology reform of chronic hepatitis to cirrhosis. It performs unbalance of extra-cellular matrix (ECM) synthesis and decomposition, which leads to deposit of ECM, and to cause fibrous connective tissue sediment abnormally in liver. Hepatic fibrosis, as an initiative and convertible procedure of hepatic injury, its pathology features are as follows, large amount of collagen fibers I and III forms in headers and lobules of liver, which leads fibers tuberosity and destroy the normal structure, which eventually leads to cirrhosis, degeneration of liver, and sometimes to be cancer. If the disease has existed without control for a long time, it will be aggravated and hepatic lobule and vessels will be rebuilt. Distances, form to pseudolobule, will appear during the sections of veins and headers, and then become to irreversible cirrhosis. In one word, hampering and converting hepatic fibrosis is one of important strategies to cure each kind of chronic hepatitis.
At present, there is lack of efficient anti-fibrosis medicines without obvious side effects, and further more, majority medicines always affect on single joint or with one target, this phenomenon impede the entire process of therapy. On the other hand, Chinese Medicine Recipes have many virtues as little cost, high security and different target points. As a result, lots of patients turn to Chinese Medicine Recipes.
The Chinese Medicine Recipe ZGG using in our research is devised by a famous Chinese physician with high reputation in Jilin Province, which is composed of Panax Ginseng C.A.Meyer, Turtles Gelatin, Radix Paeoniae Rubra Polygonummultiflorum Thunb, and Rehmannia glutinosa as basis. And what is more, it has the function of activating blood circulation, soften lump, loosen knot and tonifying energy. ZGG has already manifested good clinical effects on liver protection in cirrhosis with lower albumin. The former pharmacological research also proved that ZGG has effects of protecting liver, promoting albumin and reversing the cirrhosis forming.
With the characters of Chinese Medicine Recipes, such as unavoidable complication and uncertain active parts of components as well as many difficulties to control its quality, are resulted in the cut of medical effects and high cost of manufacture, all of these hamper the widely use and modern developments of Chinese Medicine Recipes. In order to control the qualities and quantities of Chinese Medicine Recipes, it is necessary to minimize the mutual effects of components in recipes, without therapeutic influences. It is the tendency of research on less components recipes.
Thus, we extract the main basis of ZGG–Panax Ginseng adds Turtles Gelatin to SGP (less components recipe).Through the experimental hepatic fibrosis rats established by hypodermic injection CCl4 and arachis oil, we intend to observe the index of protein level, hepatic function, oxidative damage and tissue collagen, compared with ZGG, in order to provide effective and feasible pharmacodynamic evidences in curing hepatic fibrosis and low albumin diseases with Chinese Medicine Recipes.
The results are as follows. After introgastric-infused for one month, SGP 4、8g/kg can enhance body constitution , reduce liver coefficient, and notable increase albumin (ALB) , A/G and lower globulin (GLO). Furthermore, it markedly decrease the level of aspartate aminotransferase (AST), alanine aminotransferase (ALT), bilirubin (BIL), and increase cholinesterase (CHE) of serum; Moreover, it remarkably downsize the content of malonaldehyde (MDA)and upsize the activity of superoxide dismutase (SOD) and glutathione-S transferase (GSH-ST) in liver .At the same time, it also leads to the drop of hydroxyproline (Hyp) in liver. As the results above, we can demonstrate that SGP has the same effects in enhancing and improving liver situations to ZGG. Thus, SGP may probably affect procedures of synthesis of protein in liver, especially synthesis of albumin, adjustment of olig-albuminemia and inversion of A/G, as well as prevention of collagen abnormally sediment. On the other hand, it maybe also through the mechanism of reducing lipid oxidation, cleaning free radicals and reducing necrosis to resist hepatic fibrosis.
In the end, we can draw a conclusion, despite of less components of SGP, there are identifiable effects to ZGG in conquering hepatic fibrosis. In addition, SGP can striking raise ALB of serum, which means SGP has the promising potential to apply and develop.
引文
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[13]闫颖.从“主客交”论治肝纤维化的思考[J].湖南中医杂志,2008,24(5):84.
[14]高峰,程留芳.中医药抗肝纤维化作用机制的研究进展[J].现代中西医结合杂志, 2005,14(3):395-397.
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[17]窦传斌.三七及水牛角粉在肝病低蛋白血症中应用观察[J].内蒙古中医药,2005,24(5):24-26.
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[2]刘家义.从肝的生理特性和病理特点探讨肝病用药的基本原则[J].山东中医药大学学报,2005,29(6):1-2.
[3]聂青和,周天仇.临床肝病研究十年回顾及展望[J].实用肝脏病杂志,2006,9(1):1-4.
[4]胡杨义,刘成海.肝纤维化的中医药治疗和研究值得关注[J].中西医肝病杂志,2008,18(1):1-4.
[5] Rockey Dc. Inhibition of rat hepaticlipocyte activation in culturer [J]. Hepatology, 1992,16:776-784.
[6]王华,魏伟.肝纤维化与细胞因子的关系[J].中国药理学通报,2002,18(2): 132-136.
[7]周永兴.现代肝硬化诊断治疗学[M].北京人民军医出版社出版,12.
[8]陆雄,刘平.肝纤维化过程中一个重要的病理改变—肝窦毛细血管化[J].中华肝脏病杂志,2001,9(l):53-55.
[9]李石.肝纤维化发生机制[J].中华消化杂志,1999,19(1):48-51.
[10]陈晓红.细胞因子对肝纤维化形成的影响[J].国外医学内科学分册,2001,28 (l):21-24.
[11] Albanis E, Friedman SL. Hepatic fibrosis Pathogenesis and principles of therapy[J].Clin Liver Dis,2001,5(2):315-334.
[12] Lamireau T, Desmouliere A, Bioulac-Sage P, et al. Mechanisms of hepatic fibrogenesis [J]. Arch Pediatr,2002,9(4):392-405.
[13]闫颖.从“主客交”论治肝纤维化的思考[J].湖南中医杂志,2008,24(5):84.
[14]高峰,程留芳.中医药抗肝纤维化作用机制的研究进展[J].现代中西医结合杂志, 2005,14(3):395-397.
[15]闫颖.从“主客交”论治肝纤维化的思考[J].湖南中医杂志,2008,24(5):84-86.
[16]朱家谷.浅议现代中药复方制剂[J].中国中药杂志,2007,32(22):2249-2252.
[17]窦传斌.三七及水牛角粉在肝病低蛋白血症中应用观察[J].内蒙古中医药,2005,24(5):24-26.
[18]雷力民,姜婷婷,林寿宁.中医药抗肝纤维化作用机理的研究进展[J].广西中医药,2007,30(6):311-313.
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