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醫(yī)學(xué)免費(fèi)論文:快速篩選調(diào)節(jié)肝脂酶活性中藥成分方法的研究
【摘要】 目的 研究快速篩選調(diào)節(jié)肝脂酶活性中藥成分的方法。方法 利用大鼠肝組織勻漿和比色法檢測肝脂酶(HL)活性，觀察不同實(shí)驗(yàn)條件及中藥成分對 HL活性的影響。結(jié)果 正常大鼠肝組織勻漿具有明顯的HL酶促動力學(xué)特性，其HL脂解活性有明顯Ca2+依賴性，受鈣調(diào)蛋白拮抗劑鹽酸異丙嗪抑制，符合純化HL的酶促動力學(xué)特性;篩選得到齊墩果酸和人參皂苷Rb2 2個(gè)增強(qiáng)肝脂酶活性成分。結(jié)論 首次用大鼠肝組織勻漿和脂酶比色測定法建立了一種簡便易行、快捷的調(diào)節(jié)肝脂酶活性中藥有效成分篩選方法，可用于影響肝脂酶活性的中藥調(diào)脂活性成分的篩選，并從有效方藥組分中篩選得到有增強(qiáng)HL活性的組分。
【關(guān)鍵詞】 肝脂酶;快速篩選;大鼠肝勻漿;復(fù)方貞術(shù)調(diào)脂方;肝脂酶激活劑
Rapid screening of active ingredients from Chinese medicine regulating hepatic lipase activitiesBEI Weijian，GUO Jiao，HU Yinming，HUANG Lihua，HU Xuguang，CAO Yang醫(yī).學(xué)全.在.線網(wǎng)站gydjdsj.org.cn
(Key Research Center of Liver Regulating for Hyperlipidemia，SATCM/ Class Ⅲ Laboratory of Lipid Metabolism， SATCM/ Academy of Tradational Chinese Medicine,Guangdong Pharmaceutical College，Guangzhou,Guangdong 510006,China)Abstract:Objective To study the new approach for rapid screening of active ingredients from Chinese medicine regulating hepatic lipase activities.Methods Rat liver homogenate and colorimetric assay of hepatic lipase (HL) activities were used to investigate the effects of different components of Chinese medicine on hepatic lipase activities.Result It was found that liver homogenate of SD rats demonstrated significant enzyme kinetic properties of HL. Its lipolytic activities was significantly Ca2+dependent,and inhibited by calmodulin antagonist promethazine hydrochloride,consistent with the enzymatic kinetic properties of purified HL reported in the literature. Oleanolic acid and ginsenosides Rb2 was screened out as the activators of HL.Conclusion The method for rapid screening of the active ingredient of Chinese medicine regulating HL activities by using rat liver homogenate in combination with colorimetric assay of HL activities was developed,which was simple and reliable,and could be used in .rapid screening of lipidlowering ingredients in traditional Chinese medicine having effects on HL activities,and screening of active ingredients that might increase HL activities from effective recipes of traditional Chinese medicine.
Key words:hepatic lipase (HL) ; rapid screening; rat liver homogenate; Fufang Zhenzhu Tiaozhi Fang(FTZ) recipe;hepatic lipas activator
肝脂酶(hepatic lipase,HL)是重要的脂代謝酶，其合成、分布、活性的變化和高脂血癥、脂肪肝、動脈粥樣硬化等密切相關(guān)。肝組織和血漿中HL酶活性的降低，可引起高脂血癥[1-3]，并在脂肪肝發(fā)病中起重要作用[4，5]。
復(fù)方貞術(shù)調(diào)脂方(簡稱FTZ)由女貞子、丹參、三七、黃連等中藥組成，含有齊墩果酸、三七皂苷、黃連素、丹參素等活性成分，在臨床應(yīng)用和實(shí)驗(yàn)研究中發(fā)現(xiàn)該方不僅具有明顯的降低總膽固醇(TC)、三酰甘油(TG)、低密度脂蛋白膽固醇(LDLC)及升高高密度脂蛋白膽固醇(HDLC)的作用,并能升高血漿HL活性、抑制動脈粥樣硬化(AS) 形成、抑制脂肪肝形成等作用[6，7]，還能使飲食性高脂血癥模型大鼠降低的肝組織和血漿中HL的活性恢復(fù)至正常水平[7]。
為了研究中藥降血脂的物質(zhì)基礎(chǔ)和篩選新藥，本研究以HL為靶點(diǎn)，以FTZ組分為調(diào)節(jié)HL活性成分的篩選對象，研究建立一種采用大鼠肝勻漿比色法測定肝脂酶活性，快速篩選調(diào)節(jié)HL活性中藥有效成分的方法，現(xiàn)報(bào)道如下。
