基于NLRP_(3)-神经递质通路初步探究知母总皂苷对肥胖小鼠脂肪组织的调控机制

Preliminary exploration of the regulatory mechanism of the saponins from Anemarrhena asphodeloides Bge.on adipose tissue of obese mice based on the NLRP_(3)-neurotransmitter pathway

观察知母总皂苷(saponins from Anemarrhena asphodeloides Bge.,SAa B)对C57BL/6J肥胖小鼠的干预作用,探讨其通过NLRP_(3)-神经递质通路对肥胖小鼠脂肪组织的调控机制。采用高脂饲料喂养7周建立小鼠肥胖模型,将造模成功的小鼠随机分为阳性药氯吉兰组(3 mg/kg)、SAa B低、中、高剂量组(30、60、90 mg/kg)和模型组,另设正常组,腹腔注射给药,每日1次,连续7周,正常组与模型组仅给予等量生理盐水。期间测定小鼠体质量,末次给药后测定小鼠腹部脂肪重量、空腹血糖、血清中总胆固醇(TC),甘油三酯(TG),高、低密度脂蛋白胆固醇(HDL-C、LDL-C)水平,小动物CT活体成像技术观察小鼠下腹部脂肪分布,HE染色观察小鼠脂肪组织病理形态学变化,Real-Time PCR法检测小鼠脂肪组织中生长分化因子3(GDF_(3))、天冬氨酸蛋白水解酶1(Caspase-1)、白细胞介素-1β(IL-1β)mRNA表达,Western blot法检测小鼠脂肪组织中核苷酸结合寡聚化结构域样受体家族热蛋白结构域3(NLRP_(3))、单胺氧化酶A(MAOA)蛋白表达。结果显示SAa B可降低肥胖小鼠体质量,减少腹腔内壁脂肪组织堆积,降低空腹血糖值、血清TC、TG、LDL-C水平和升高HDL-C水平,改善脂肪组织病理状态,抑制小鼠脂肪组织中GDF_(3)、Caspase-1、IL-1βmRNA表达及NLRP_(3)、MAOA蛋白表达。本研究结果表明SAa B能降低高脂饮食诱导的肥胖小鼠体质量,控制血糖血脂代谢紊乱,减轻炎症反应,促进肥胖小鼠脂类分解,减少腹部脂肪蓄积,其机制可能与SAa B调控NLRP_(3)-神经递质通路有关。

To observe the intervention effect of saponins from Anemarrhena asphodeloides Bge.(SAa B)on C57BL/6J obese mice and to study the regulation mechanism of SAa B on adipose tissue of obese mice through NLRP_(3)-neurotransmitter pathway.The obesity model of mice were established by feeding with high-fat diet for 7 weeks,the successfully modeled mice were randomly divided into the clorgiline group(3 mg/kg),low-dose,medium-dose and high-dose SAa B groups(30,60 and 90 mg/kg)and the model group,in addition,set up a control group.Mice were given intraperitoneal injection once a day for 7 weeks,the control group and the model group were given only the same amount of normal saline.During this period,the body weight of mice were measured.After the last administration,the weight of abdominal fat,fasting blood glucose and the levels of total cholesterol(TC),triglyceride(TG),high and low density lipoprotein cholesterolserum lipids(LDL-C,HDL-C)were measured.The distribution of lower abdominal fat was observed by CT in vivo imaging technique.The pathomorphological changes of mice adipose tissue were observed by hematoxylin-eosin(HE)staining and the mRNA expression of growth differentiation factor 3(GDF_(3)),caspartate proteolytic enzyme 1(Caspase-1)and interleukin-1β(IL-1β)in adipose tissue of mice were detected by Real-Time PCR method,the protein expression of nucleotide-binding oligomerization domain-like receptor family thermoprotein domain 3(NLRP_(3))and michigan association for openness in adoption(MAOA)in mice adipose tissue were detected by Western blot.The results show that SAa B can reduce the body mass of obese mice,the accumulation of adipose tissue in abdominal wall,decrease the levels of fasting blood glucose,serum TC,TG,LDL-C and increase the level of HDL-C,improve the pathological state of adipose tissue,and inhibit the expression of GDF_(3),Caspase-1,IL-1βmRNA and NLRP_(3),MAOA protein in adipose tissue of obese mice.The results of this study show that SAa B can reduce the body weight of obese mice induced by high-fat diet,control the disorder of blood glucose and blood lipid metabolism,reduce the inflammatory reaction,promote the decomposition of lipids and reduce the accumulation of abdominal fat in obese mice,and the mechanism may be related to the regulation of NLRP_(3)-neurotransmitter pathway by SAa B.