摘要
目的研究绞股蓝总皂苷对非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)大鼠肝脏保护作用及免疫调节作用机制。方法 48只SD大鼠随机分为正常对照组,模型对照组,多烯磷脂酰胆碱胶囊组(阳性对照组,150 mg·kg^(-1))及绞股蓝总皂苷高、中、低(240,120,60 mg·kg^(-1))剂量组,除正常对照组外,其余各组连续饲喂高脂饲料10周,制备NAFLD模型;模型成功后,各组连续给药8周。实验期间,分别于给药0,4,8周眼眶取血检测血清AST、ALT;末次给药后,采用流式细胞技术检测外周血Th17和Treg细胞含量;酶联免疫法检测血清IL^(-1)7、IL^(-1)0及TNF-α含量。HE染色检查肝组织病理变化和免疫组化法检查肝组织IL^(-1)7和Foxp3表达。结果给药4周,绞股蓝总皂苷高剂量显著降低大鼠血清ALT、AST(P<0.01);给药8周,绞股蓝总皂苷各剂量均能显著降低血清ALT、AST(P<0.05,0.01)。绞股蓝总皂苷高、中剂量能改善肝组织病变,降低TNF-α水平;高剂量显著降低IL^(-1)7、升高IL^(-1)0水平(P<0.05,0.01),降低淋巴细胞IL^(-1)7含量,升高CD4+CD25+Treg含量(P<0.05),明显减少炎症因子IL^(-1)7的表达和增加Foxp3表达。结论绞股蓝总皂苷能改善NAFLD大鼠肝脏病变,其作用机制可能与调节肝脏Treg/Th17细胞平衡,减少促炎症因子和增加抗炎因子产生相关。
OBJECTIVE To study the effect of stevenleaf on improving non-alcoholic fatty liver disease in rat model and clarify mechanism from the point of immune-regulation. METHODS Forty-eight of rats were divided into six groups： normal group, model group, Essentiale （150 mg·kg^-1）, high-dose （240 mg·kg^-1）, moderate-dose （120 mg·kg-1） and low-dose of stevenleaf （60 mg·kg-1） groups. Rats were given high fat food for continuous 10 weeks, then given correspond drugs for 8 weeks. The levels of serum AST and ALT were measured at 0, 4 and 8 weeks experimental session. The rats were anesthetized after administration 8 weeks. The peripheral blood lymphocytes and serum were separated to measure the contents of Thl7 and Treg cells in peripheral blood by flow cytometry and the levels of IL-17, IL-10 and TNF-ct in serum by enzyme-linked immunosorbent assay （ELISA）. HE staining was used to observe the pathological changes of liver tissue and immunohistochemistry to observe the positive expression of IL-17 and Foxp3 in liver tissue. RESULTS The 240 mg·kg-1 of stevenleaf could reduce levels of serum ALT and AST after administration for 4 weeks （P〈0.01）. stevenleaf 240 mg·kg-1, 120 mg·kg-1 and 60 mg·kg-1 could reduce levels of serum ALT and AST after administration for 8 weeks （P〈0.05, 0.01）. Stevenleaf 240 mg·kg-1 and 120 mg.kg-1 could improve liver damage and reduce the level of TNF-α （P〈0.05）. Stevenleaf 240 mg＇kg-1 could reduce the content of IL-17 and increase IL-10 in serum （P〈0.05, 0.01）, reduce content of Thl7 cell （CD4＋IL-17＋） and increase CD4＋CD25＋ Treg in peripheral blood lymphocytes （P〈0.05）, also significantly reduced the expression of inflammatory factor IL-17 and increased Foxp3 expression. CONCLUSION Stevenleaf could protect liver tissue damages, its mechanism may be related to regulating Treg/Th 17 cell balance, reducing nro-inflammatorv factors and increasing anti-inflammatory factors.
出处
《中国现代应用药学》
CAS
CSCD
2017年第12期1683-1688,共6页
Chinese Journal of Modern Applied Pharmacy
基金
浙江省中西医结合学会科研项目(2014LYZD001)
