摘要
目的观察石斛灌胃对非酒精性脂肪肝(NAFLD)小鼠肝细胞内脂肪过度沉积的改善作用,并探讨其机制。方法取10只C57BL/6J雄性小鼠作为正常对照组,普通饲料喂养,剩余30只小鼠用高糖高脂饲料连续饲养12周,建立NAFLD模型后随机分成模型组、石斛低剂量组、石斛高剂量组,每组10只。石斛低剂量组、石斛高剂量组分别给予200 mg/(kg·d)、400 mg/(kg·d)石斛汤剂灌胃,模型组、正常组均灌服等体积生理盐水,连续灌胃14周。灌胃结束后,观察各组小鼠体形、体质量、腹围、皮毛光泽度及活动量等总体状态。灌胃结束后,各组小鼠禁食12 h,眼下静脉丛取血,采用生化测试试剂盒检测血清TG、TC、AST、ALT。灌胃结束后处死小鼠,取肝脏组织,伊红-苏木素染色后镜下观察肝脏组织病理。采用PT-PCR法检测各组小鼠肝脏组织中固醇调节元件结合蛋白(SREBP-1c)mRNA、脂肪酸合成酶(FAS)mRNA。采用Western Blotting法检测各组小鼠肝脏组织中p-mTOR蛋白。结果模型组小鼠体形明显大于正常组,体质量和腹围也较正常组明显增加,动作自如,但皮毛粗糙失去光泽且易脱毛。石斛低剂量组、石斛高剂量组小鼠活动量正常,活动灵敏,精神正常,皮毛光泽。正常组、石斛低剂量组、石斛高剂量组小鼠血清TG、TC、AST、ALT水平均低于模型组(P均<0.05)。模型组小鼠肝细胞肿大,脂滴较多,并有脂肪囊肿形成;石斛低、高剂量组小鼠肝脏中脂肪肝转态减轻。正常组、石斛低剂量组、石斛高剂量组小鼠肝脏组织中SREBP1c mRNA、FAS mRNA相对表达量均低于模型组(P均<0.05),且石斛高剂量组小鼠肝脏组织中SREBP1c mRNA、FAS mRNA相对表达量均低于石斛低剂量组(P均<0.05)。正常组、石斛低剂量组、石斛高剂量组小鼠肝脏组织中p-mTOR蛋白相对表达量均低于模型组(P均<0.05),且石斛高剂量组小鼠肝脏组织中pm TOR蛋白相对表达量低于石斛低剂量组(P均<0.05)。结论石斛灌胃可改善NAFLD小鼠的肝细胞内脂肪过度沉积,其机制可能与参与m TOR信号通路的调节有关。
Objective To observe the effect of dendrobium on the amelioration of excessive fat deposition in the liver cells of non-alcoholic fatty liver disease(NAFLD)mice,and to explore its mechanism.Methods Ten C57 BL/6 J male mice were taken as the normal control group and fed with ordinary diet.The remaining 30 mice were fed with high-sugar and high-fat diet for 12 weeks.After the NAFLD models were established,they were randomly divided into the model group,low-dose and high-dose dendrobium groups,with 10 mice in each group.The mice in the low-dose and high-dose dendrobium groups were given 200 and 400 mg/(kg·d)dendrobium decoction by gavage,respectively.Both the mice in the model group and the normal group were given equal volume of normal saline for 14 weeks.After the treatment,the status of the mice in each group,including body shape,weight,abdominal circumference,fur gloss,and activity level were observed.After the gavage,the mice in each group were fasted for 12 h,and blood was taken from the venous plexus under the eyes,and the serum TG,TC,AST and ALT were detected with a biochemical test kit.After the gavage,the mice were sacrificed,and the liver tissues were taken to observe the liver tissue pathology.PT-PCR was used to detect sterol regulatory element binding protein(SREBP-1 c)mRNA and fatty acid synthase(FAS)mRNA in the liver tissues of mice in each group.Western blotting was used to detect the p-mTOR protein in the liver tissues of mice in each group.Results The body shape of the mice in the model group was significantly larger than that of the normal group,and their weight and abdominal circumference also significantly increased.They moved freely,but their fur was rough and dull and easy to shed hair.The mice in the low-dose and high-dose dendrobium groups had normal activity,agile activity,normal spirit,and shiny fur.The serum TG,TC,AST and ALT levels of mice in the normal group,low-dose and high-dose dendrobium groups were lower than those in the model group(all P<0.05).Mice in the model group had enlarged liver cells,more lipid droplets,and fatty cyst formation;the fatty liver transformation decreased in liver of the mice in the low-dose and high-dose dendrobium groups.The relative expression levels of SREBP1 c mRNA and FAS mRNA in the liver tissues of mice in the normal group,low-dose and high-dose dendrobium groups were lower than those in the model group(all P<0.05),and the relative expression of SREBP1 c mRNA and FAS mRNA of liver tissues in high-dose Dendrobium group was lower than that of the low-dose dendrobium group(all P<0.05).The relative expression of p-mTOR protein in the liver tissue of mice in the normal group,low-dose and high-dose dendrobium group was lower than that in the model group(all P<0.05),and the relative expression level of p-mTOR protein in the liver tissues of the high-dose dendrobium group was lower than that of the low-dose dendrobium group(P<0.05).Conclusion Dendrobium may regulate the m TOR signaling pathway to ameliorate the excessive fat deposition in the liver cells of NAFLD mice.
作者
梁钰华
杨育辉
邓晓冬
林成创
LIANG Yuhua;YANG Yuhui;DENG Xiaodong;LIN Chengchuang(Guangzhou Panyu Central Hospital,Guangzhou 511400,China)
出处
《山东医药》
CAS
2020年第21期48-52,共5页
Shandong Medical Journal
基金
广东省中医药局科研项目(20191264)。
关键词
石斛
非酒精性脂肪肝
肝细胞
m
TOR信号通路
dendrobium
non-alcoholic fatty liver
liver cells
m TOR signaling pathway
