华蟾素致泻机制初步研究  

A preliminary study on the mechanism of catharsis induced by cinobufagin

作　　者：孙丽丽 罗川 沈龙海[1] 潘绵立[1] SUN Li-li;LUO Chuan;SHEN Long-hai;PAN Mian-li(State Key Laboratory of New Drug and Pharmaceutical Process,Shanghai Institute of Pharmaceutical Industry,China State Institute of Pharmaceutical Industry,Shanghai Professional and Technical Service Center for Biological Material Drug Gability Evaluation,Shanghai 200437,China)

机构地区：[1]中国医药工业研究总院上海医药工业研究院创新药物与制药工艺国家重点实验室,上海市生物物质成药性评价专业技术服务中心,上海200437

出　　处：《中国新药杂志》2023年第22期2311-2316,共6页Chinese Journal of New Drugs

摘　　要：目的:研究华蟾素原液及其主要成分生物碱对大鼠体外结肠平滑肌收缩的影响、体内引起腹泻的相关通道蛋白异常表达的影响。方法:取正常大鼠部分结肠组织标本放入浴槽中连接移位张力换能器,待结肠标本张力稳定后给予不同剂量的华蟾素、生物碱溶液,记录结肠标本收缩张力变化。给予大鼠等效临床剂量华蟾素及生物碱溶液,连续灌胃给药7 d,d 8处死大鼠,取结肠组织,采用Western blot检测钙通道蛋白CaV1.2和CaV1.3、钠通道蛋白NHE2和NHE3、水通道蛋白AQP3和AQP8的表达变化。结果:原液<0.10 mg(含药量<0.09 mg),结肠标本张力无明显变化;原液0.10~18.74 mg(含药量0.09~16.21 mg),结肠标本张力变化增加;原液>19.25 mg(含药量>16.64 mg),结肠标本张力降低。生物碱<0.06μg,结肠标本张力无明显变化;生物碱0.06~23.41μg,结肠标本张力增加;生物碱>24.03μg,结肠标本张力降低。与正常组相比,华蟾素组NHE2,NHE3,AQP3,AQP8显著降低(P<0.05),CaV1.2显著增加(P<0.05);生物碱组NHE3和AQP3显著降低(P<0.05)。其余蛋白表达均无显著性差异(P>0.05)。结论:致泻机制研究中,离体结肠张力实验的结果表明,一定剂量的华蟾素可促进肠蠕动,提高腹泻的发生率;高于一定剂量后可抑制肠蠕动。结肠中腹泻相关蛋白水平检测结果表明,给予华蟾素和生物碱溶液均会引起腹泻相关通道蛋白的变化,生物碱溶液对通道蛋白产生显著性影响的种类少于原液,提示致泻成分除生物碱外还有其他成分。Objective:To study the effects of cinobufagin and its main component alkaloids on the contraction of colonic smooth muscle in rats in vitro and their influence on abnormal expression of channel proteins related to diarrhea in vivo.Methods:Normal rat colon tissue samples were put into the bath to connect with the displacement tension transducer.After the tension of the colon sample was stabilized,different doses of cinobufagin and alkaloid solution were given to the colon sample,and the changes of the contraction tension of the colon sample were recorded.The rats were given the clinically equivalent dose of cinobufagin and alkaloid solution by gavage for 7 consecutive days,and the rats were killed on the 8th day.Colon tissues were taken,and the expressions of calcium channel proteins CaV1.2,CaV1.3,sodium channel proteins NHE2,NHE3,aquaporins AQP3,AQP8 were detected by Western blot to observe the changes.Results:The tension of colon specimen did not change significantly when the original solution was<0.10 mg(drug content<0.09 mg);0.10~18.74 mg stock solution(0.09~16.21 mg)increased the tension of colon specimen.If the original solution was>19.25 mg(drug content>16.64 mg),the tension of colon specimen decreased.When alkaloids was<0.06μg,the tension of colon specimen had no obvious change;0.06~23.41μg alkaloid increased the tension of colon specimen.When alkaloids was>24.03μg,the tension of colon specimen decreased.Compared with the normal group,the sodium channel proteins NHE2,NHE3 and aquaporins AQP3,AQP8 in the cinobufotalin group decreased significantly(P<0.05),while the calcium channel protein CaV1.2 increased significantly(P<0.05).Sodium channel protein NHE3 and aquaporin AQP3 were significantly decreased in the alkaloid group(P<0.05).There was no significant difference in the expression of other proteins(P>0.05).Conclusion:In the study of diarrhea mechanism,the results of colon tension test in vitro showed that a certain dose of cinobufagin could promote intestinal peristalsis and increase the incidence of

关 键 词：华蟾素 腹泻 生物碱 

分 类 号：R965[医药卫生—药理学]