目的 探究波棱瓜子壳总甾醇（HCSTS）对小鼠急性肝损伤保护作用及机制，并对两个主要甾醇进行含量测定。方法 60只雄性ICR小鼠随机分成空白组（CON）、肝损伤组（MODEL）、水飞蓟素组（SIL）、HCSTS低（LD）、中（MD）、高剂量组（HD），通过腹腔注射0.3% CCl₄-橄榄油诱导急性肝损伤。造模24 h后，采血用于丙氨酸氨基转移酶（Alanine aminotransferase， ALT）和天门冬氨酸氨基转移酶（Aspartate aminotransferase， AST）活力检测，取肝组织用于组织病理观察，qRT-PCR测定肝组织炎症相关指标IL-1β、IL-6、IL-10和环氧合酶-2（Cyclooxygenase-2， COX-2）转录水平。RP-HPLC法用于HCSTS中两个主要甾醇的含量测定。结果 与MODEL相比，MD、HD组能显著减少肝组织炎性细胞浸润，降低血清ALT、AST活力。HD组能显著降低IL-1β、IL-6和COX-2的转录水平（P<0.05），并能显著升高IL-10的转录水平（P<0.05）；反相高效液相色谱法测得两个主要甾醇7，22，25-豆甾三烯醇和α-菠甾醇含量分别为20.4%和54.2%。结论 HCSTS对CCl₄诱导的急性肝损伤有保护作用，其作用机制可能与减少IL-1β、IL-6的转录，增加IL-10的转录，并降低COX-2的表达有关。含量测定结果表明，7，22，25-豆甾三烯醇和α-菠甾醇可能是HCSTS抗肝损伤的两个主要活性成分。

Objective To study the hepatoprotective effects and potential mechanism of total sterols extracted from the shell of Herpetospermum caudigerum Wall. (HCSTS) on mice with acute liver injury induced by CCl₄, and develop a RP-HPLC method for simultaneous quantification of two major ingredients.Methods 60 mice were randomly divided into control group (CON), model group (MODEL), silymarin group (SIL), HCSTS-low (LD), medium (MD) and high dose group (HD), and the mice were injected intraperitoneally with 0.3% CCl₄ - olive oil solution to induce the acute liver injury except the control group. 24h later, the eyeballs of mice were removed to take blood, and all mice were sacrificed to collect the liver tissues. Biochemical methods were used to measure the serum contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Quantitative-real time-reverse transcription (qRT-PCR) was used to analyze the gene expression of inflammatory-related indicators, such as interleukin-1β (IL-1β), interleukin-10 (IL-10) and cyclooxygenase-2 (COX-2). HE staining was applied to observe the histopathological changes of liver. A RP-HPLC method for the quantification of two major ingredients in HCSTS was developed.Results Compared with the MODEL group, the results showed that MD and HD groups of HCSTS could significantly improve the liver tissue lesion, decrease the serum contents of AST and ALT, enhance the gene expression of IL-10 (P<0.05), and inhibit the gene expression of IL-1β, IL-6 and COX-2 (P<0.05). The contents of the two major ingredients 7,22,25-Stigmatrienol and α-Spinasterol were 20.4% and 54.2%.Conclusion HCSTS has a protective effect on mice with acute liver injury induced by CCl₄, and its mechanism may be related to inhibiting the gene expression of IL-1β, IL-6 and COX-2, enhancing the gene expression of IL-10. As the result of quantification, it could be inferred that 7,22,25-Stigmastatrienol and α- Spinasterol were major active ingredients which protected against CCl₄-induced liver injury.

R285.5

中国藏学研究中心项目（1171021448）：藏药波棱瓜生长机理探索及多元利用合作研究