摘要
目的:观察中药别甲、大贝等复方肝纤克对实验性肝损伤及肝纤维化的保护作用。方法:①实验于2004-03/2005-01在三峡大学天然药物重点实验室完成。选用昆明系小鼠90只,Wistar大鼠90只,雌雄不拘。肝纤克主要成分为别甲、大贝、玳瑁、蚤休、田七、白花蛇等,由三峡大学医学院药理学教研室自制。②观察肝纤克对硫代乙酰胺致小鼠肝纤维化的影响:取昆明系小鼠30只,随机分为3组:对照组:以生理盐水0.2mL/只,灌胃;肝纤克低和高剂量治疗组:以100g/L肝纤克0.2和0.5mL/只,灌胃,1次/d,共3d。于末次给药后24h,各鼠均腹腔注射硫代乙酰胺50mg/kg,并如上法重复给药1次。再过24h后取小鼠眼眶血,以赖氏法测定血清谷丙转氨酶水平。③观察肝纤克对氨基半乳糖致小鼠肝纤维化的影响:取昆明系小鼠30只,分组及组名同上。然后腹腔内注射D-氨基半乳糖500mg/kg。72h后取小鼠眼眶血,以赖氏法测定谷丙转氨酶水平。④观察肝纤克对四氯化碳致小鼠肝纤维化的影响:取昆明系小鼠30只,分组及组名同上。末次给药后24h,各鼠腹腔注射1g/L四氯化碳石腊油10mL/kg,并如上法再给药1次。24h后取小鼠眼眶血,以赖氏法测定谷丙转氨酶水平。⑤观察肝纤克对四氯化碳致大鼠慢性肝纤维化的影响:取Wistar大鼠90只,随机分为6组:空白对照组(给予正常饮食,不给任何药物);盐水对照组:造成肝纤维化模型后[以1g/L四氯化碳石蜡油(0.2mL/只)作背部皮下注射,每隔3d1次,共45d,12次进行造模],以生理盐水进行干预;预防性肝纤克低和高剂量治疗组:造成肝纤维化模型的同时,分别用0.5和1.0g/kg肝纤克,灌胃进行预防性治疗,1次/d;肝纤克低和高剂量治疗组:造模后,分别用0.5和1.0g/kg肝纤克进行治疗。治疗各组均用肝纤克干预60d。于末次注射1g/L四氯化碳石蜡油24h后,从盐水对照组中随机取2只大鼠处死,做肝脏组织切片,以检查成型情况。肝纤克干预60d后,自大鼠尾部取血,检查血清谷丙转氨酶水平,并以对硝基磷酸酚动态法血清碱性磷酸酶水平。统计肝纤维化的发生率。⑥根据组织形态学变化,将肝纤维化分为轻度(汇管区结缔组织增生,并轻度向小叶内伸展,汇管区因而增宽,伴少量慢性炎性细胞浸润),中度(汇管区有明显的纤维结缔组织增生,并较明显地向小叶内伸展,伴较多慢性炎性细胞浸润,肝小叶结构被重新划分,但尚无典型假小叶形成),重度(除上述表现加重外,有典型假小叶形成)。⑦计量和计数资料差异比较分别采用t检验和χ2检验。结果:小鼠90只和大鼠90只均进入结果分析。①肝纤克低剂量和高剂量治疗组硫代乙酰胺、氨基半乳糖、四氯化碳致肝损伤小鼠血清谷丙转氨酶水平明显低于对照组(P<0.05,0.01)。②肝纤克治疗各组及空白对照组四氯化碳致慢性肝纤维化大鼠血清谷丙转氨酶和碱性磷酸酶水平均明显低于盐水对照组(P<0.05～0.01),肝纤克治疗各组接近空白对照组。预防性肝纤克低和高剂量治疗组肝纤维化发生率均明显低于盐水对照组(P<0.01),肝纤维化程度低,多为轻度。肝纤克低和高剂量治疗组与盐水对照组接近,多呈肝纤维化重度表现。结论:①肝纤克对硫代乙酰胺致小鼠急性肝损伤有一定的保护作用。②当预防性给药时,肝纤克既可保护肝脏的功能又可抑制肝纤维化的形成,而当肝纤维化形成后治疗性给药时,肝纤克虽能保护肝脏的功能,但却不能消除或缓解已经形成的纤维化变化。
AIM： To evaluate the protective effect of Gan Qian Ke （GQK） on liver injury and hepatic fibrosis in experimental animals.
METHODS： （1）This experiment was conducted at Key Laboratory of Natural Drugs, Throe Gorges University between March 2004 and January 2005. Ninety Kunming mice and 90 Wistar rats at different sex were selected. The GQK （main components： Biejia, Dabei, eretmoohelys, Zaoxiu, Tianqi, Agkistrodon acutus, and so on） was made by Staff Room of Pharmacology, Medical College, Three Gorges University. （2）To evaluate the effect of the GQK on mouse hepatic fibrosis induced by thioacetamide： Thirty Kunming mice were selected and assigned into three groups randomly： The mice in the control group were given saline with gastric perfusion （0.2 ml each mouse）, 100 g/L GQK low dose group（0.2 ml each mouse）, 100 g/L GQK high dose group （0.5 ml each mouse）, with gastric perfusion, once a day for 3 days. After the last administration 24 hours, thioacetamide 50 mg/kg was injected into every mouse through intraperitoneal injection and repeat dosage as above. Fossa orbitalis blood was got, and level of blood serum glutamic-pyruvic transaminase （GPT） was detected with Reitman method after 24 hours. （3）To evaluate the effect of GQK on mouse hepatic fibrosis induced by galactosamine： Thirty Kunming micewere selected and assigned into groups with the same name as above. 500 mg/kg D-galactosamine were injected with intraperitoneal injection. The fossa orbitalis blood was got, and the level of blood serum GPT was detected with Reitman method after 72 hours. （4）To evaluate the effect of GQK on mouse hepatic fibrosis induced by Tetrachloromathane： Thirty Kunming mice were selected and assigned into groups with the same name as above. After last administration 24 hours,1 g/L tetraehloromathane paraffin 10 ml/kg was injected into every mouse through intraperitoneal injection and repeat dosage as above. The fossa orbitalis bloodwas got, and the level of blood serum Girl, was detected with Reitman method after 24 hours. （5）To evaluate the effect of GQK on rat chronic hepatic fibrosis induced by tetrachloromathane： Ninety Wistar rats were selected and randomly divided into 6 groups： blank control group （normal diet and never to give any drugs）;, saline control group, after establishing hepatic fibrosis models [1 g/L tetrachloromathane paraffin （0.2 ml each mouse） with subcutaneous injection at back, once every 4 days for 45 days, 12 times], intervening with saline; prophylactic GQK low and high dose groups, 0.5 g/L and 1.0 g/L GQK was given to rats, respectively at equal pace of the model to succeed, once a day; GQK low and high dose groups, 0.5 g/L and 1.0 g/L GQK was given to rats, respectively at equal pace of the model to succeed for 60 days. After 24 hours last hypodermic injection with 1 g/L tetrachloromathane paraffin, two rats from saline control group gained randomly were sacrificed. Liver histological section was commited for checking out the condition of models. After intervening with GQK for 60 days, tail blood of rats was gotten, and to detect the level of serum GPT, and the level of serum alkaline phosphatase （AKP） was detected with obnitrophosphophenol dynamic method. Incidence rate of liver hepatic fibrosis was added up. （6）According to the histological and morphological changes, liver hepatic fibrosis was separated into light （desmoplasia in portal area and verge to hepatic lobules lightly, portal area was widen and to accompany with small amounts of inflammatory cell infiltration）, middle （obvious desmoplasia in portal area and verge to hepatic lobules markedly, portal area was widen and to accompany with large amounts of inflammatory cell infiltration, the framework of hepatic lobules had been divla renewedly, but representative abnormal hepatic lobules to shape not yet）, severity （apart from above-mentioned, it also had representative abnormal hepatic lobules to shape）. （7）Measurement data and numeration data were compared with t-test and A3 test, respectively.
RESULTS： Ninety mice and 90 rats were all involved in the result analysis. （1）The serum GPT levels of mice hepatic fibrosis induced by thioacetamide , D-galactosamine and tetrachloromathane were lower markedly than those in the control group （P 〈 0.05,0.01 ）. （2）The serum GPT and AKP levels of rats chronic hepatic fibrosis induced by tetrachloromathane were remarkably lower than those in the saline control groups （P 〈 0.05-0.01 ）, and the levels in the GQK treatment group were closed to those in the blank control group. Hepatic fibrosis incidence rates of prophylactic GQK low and high dose groups were remarkably lower than those in the saline control groups（P 〈 0.01 ）. The levels of hepatic fibrosis were low, and most of it was light. It was close between the GQK low and high dose groups and the saline control group, and most were severe hepatic fibrosis.
CONCLUSION： （1）GQK have protective effect on mouse hepatic fibrosis induced by thioacetamide. （2）When performing prophylactic administration, GQK can not only protect liver function, but also inhibit the formation of hepatic fibrosis. However, after the formation of hepatic fibrosis, when giving remedial drugs the GQK Can protect the hepatic function, but cannot preclude or relieve the changes of fibrosis.
出处
《中国临床康复》
CSCD
北大核心
2006年第3期96-98,共3页
Chinese Journal of Clinical Rehabilitation
