益肝康药物血清对大鼠肝星状细胞胞浆游离钙效应的抑制作用.pdf

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论著
益肝康药物血清对大鼠肝星状细胞胞浆游离钙效应的扣制作用
王天轶,王军民,王如美,柳继兴,姚希贤
【摘要】目的研究益肝康药物血清是否具有抑制肝星状细胞(HSCs)胞浆游离钙([Ca2+]i)增高的作用。
方法将32只健康雄性SD大鼠接随机数字表分为4组:肝纤维化模型益肝康组(8只)皮下注射用精制橄榄油配制
的40%CCL4溶液9周,然后灌服益肝康6d:肝纤维化模型对照组(8只)皮下注射用精制橄榄油配制的40%CCL与溶
液9周,然后灌服0.9%氯化钠溶液6d;正常大鼠益所康组(8只)皮下注射等量精制橄榄油9周,然后灌服益肝康
6d;正常对照銀(8只)皮下注射等量精制橄榄油9周,然后灌服0.9%氯化钠溶液6d。结東后经下腔静脉取血并分
离血清。用言法,采用上述10%血清培养HSCs24h并负載好uo-3/AM后,使用激光扫描共聚焦显微镜(LSCM)
检测HSCs[Ca2+]i。结果(1)经正常大鼠益肝康组及肝纤維化模型益肝康組大鼠血清预处理的HSCs,其
Ca2+]i荧光强度相对值均明显低于肝纤维化模型对照组(P<0.05);且两者与正常对照组间差别均有显著性意义
(P<0.05)。结论益肝康能抑制肝星状细胞[Ca2+]i的升高,这可能是其发挥抗肝纤維化作用的重要途径之一。
【关键词】肝纤维化;肝星状细胞;益肝康;激光扫描共聚焦显微铣;胞浆游高钙
【中图分类号】R575.2R285.5【文献标识码】A【文章编号】1007-9572(2006)14-1172-03
Effects of Yigankang on Ca* in the Course of HSCS Activiting WANG Tian
WANG Jun-min, WANG Ru-mei
et al. Department of Emergency, Bai qiuen International Peace Haspital, Shijiazhuang 050000, China
(Abstract) Objective To investigate whether pharmacological serum from the rats taking Yigankang can inhibit the in-
crease of intracellular free calcium level in HSCS. Methods 3 health male rats were divided into4 groups, hepatic fibrosis
group was injected by 40 Ls subcutaneously for 9 weeks. Then Yigankang was filled into the stomachs of one medicated
group but the same volume of O.9% Nacl into another medicated groups twice a day for 6 consecutive days. The same thing hap-
pened in another two groups which were not modeled into hepatic fibrosis by 40% CCL4. The pharmacological serum
rawn
from the inferior vena cava in two hours after the last time filling of drugs or Nacl on the morning of the 7th day. The serum drew
rom rats was ispensed n the 8 RPMI-1640 cul ue meim o make the 10% ug em Rpmi1640cul ure medium mix
ture, which was used to cultivate HSCS, Fluo-3-loaded HSCS and examined effects of Yigankang on [ Ca*] i with laser
scanning confocal microscopy (CM). Results In activated HSCS, both Yigankang pharmacological serums decreased
Ca2*] i in HSCS significantly compared with controls (P <0. 05). Conclusion Yigankang decreased [Ca*] i in activa-
ted HSCS, which may be one of important ways to block liver fibrosis
Key word) Liver fibrosis(LF); Hepatic stellate cells(HSCS); Yigankang; Laser scanning confocal microscopy
( LSCM): Intracellular free calcium([ Ca2+]i)
肝纤维化是慢性肝病进展成肝硬化的共同病理基础与必经焦显微镜( laser scanning confocal microscopy,LSCM),采用第
阶段,而肝星状细胞( hepatic stellate cells,HSCs)的活化与三代钙荧光探针uo-3/AM技术,观察益肝康对HSCs胞浆
增殖则是肝纤维化发生的中心环节,因此抑制HSCs活化与增游离钙([Ca2+]i)荧光强度变化的影响,探讨其抑制HSCs
殖则成为阻止并逆转肝纤维化的主要途径之一。而细胞内活化与增殖的机制。
Ca2'水平的升高在HSCs的活化与増殖中发挥重要作用(.1。1材料与方法
目前大量研究结果已证实包括益肝康( Hirude)在内的许多中1.1实验动物雄性清洁级SD大鼠32只,体重180~250g,
药对脏器纤维化有一定的治疗作用,。然而尚乏益肝康对由河北医科大学实验动物中心提供。
HSCs细胞内Ca2+信号转导机制的研究,本研究借助激光共聚
2细胞株肝星状细胞株CFSC由美国 Greenwel教授建株
基金项目:河北省科技厅基金资助项目(02276103D-8)
并惠難,其表型为活化的HSCs,从四氯化碳(CCL4)诱发的
肝硬化大鼠中分离并通过培养使细胞自发获得永生性,冷冻保
作者单位:050000河北省石家庄市,白求恩国际和平医院急诊科
(王天铁);河北省人民医院消化科(王军民);河北工程大学医学院
存于液氮中。
(王如美,柳继兴);河北医科大学附属第二医院消化科(姚希贤)
L.3主要仪器及试剂ISCM购自德国 Leica公司。AngI购
自美国 Sigma公司,益肝康购自石家庄市乐仁堂,RPMI