熟地黄水提液通过外泌体miR-29a-3p调节OVX大鼠的脂代谢

Water extraction of Rehmanniae Radix Praeparata regulates lipid metabo⁃lism via exosomal miR-29a-3p in OVX rats

目的:探讨熟地黄(Rehmanniae Radix Praeparata,RR)水提液对双侧卵巢切除(ovariectomy,OVX)大鼠脂代谢紊乱的影响及作用机制。方法:将40只3月龄SD雌性大鼠随机分为5组,分别为假手术组、OVX组、RR水提液低剂量组、RR水提液中剂量组和RR水提液高剂量组,对大鼠分别行假手术或OVX,药物灌胃12周后取材,测定大鼠的体重、腹围、血清雌二醇(estradiol,E2)、子宫重量、胰岛素(insulin,INS)和血脂四项;行肝脏HE染色和油红O染色观察脂质积累;行腹部脂肪HE染色观察脂肪体积;Western blot和RT-qPCR检测腹部脂肪过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和白细胞介素1β(interleukin,IL-1β)的表达;RT-qPCR检测肥胖相关因子糖蛋白非转移性黑色素瘤蛋白B(glycoprotein non-metastatic melanoma protein B,GPNMB)、血清外泌体miR-29a-3p、腹部脂肪miR-29a-3p及其靶基因核因子I A(nuclear factor I A,NFIA)的表达;Western blot检测腹部脂肪Wnt信号通路相关蛋白的表达。结果:RR水提液显著降低OVX大鼠的腹围、INS、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、肝脏脂质积累、腹部脂肪细胞扩张、PPARγ、TNF-α、IL-1β、GPNMB和NFIA的表达,显著升高OVX大鼠的血清E2、血清外泌体miR-29a-3p和腹部脂肪miR-29a-3p水平,激活腹部脂肪Wnt信号通路。结论:RR水提液可降低OVX大鼠的血清E2、INS、LDL-C和肝脏脂质积累,抑制腹部脂肪细胞扩张和炎症反应,其作用机制可能是与血清外泌体中miR-29a-3p和腹部脂肪中miR-29a-3p的含量增加后致NFIA表达量下降,从而激活腹部脂肪组织中Wnt信号通路有关。

AIM:To investigate the effect of water extract of Rehmanniae Radix Praeparata(RR)on lipid metabolism in OVX rats.METHODS:Forty 3-month-old SD female rats were randomly divided into sham group,OVX group,RR-L group,RR-M group and RR-H group.The body mass,abdominal circumference,serum E2,uterine weight,insulin(INS),and the level of blood lipid of the rats were detected 12 weeks later.HE staining and Oil red O staining for livers were used to observe lipid accumulation.HE staining was used to evaluate the lipocyte size of abdominal adipose.The expression of peroxisome proliferator-activated receptorγ(PPARγ),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),glycoprotein non-metastatic melanoma protein B(GPNMB)in abdominal adipose,miR-29a-3p in serum exosome and abdominal adipose,together with its target gene nuclear factor I A(NFIA)were measured by RT-qPCR and/or Western blot.Wnt signaling pathway in abdominal adipose was detected by Western blot.RESULTS:Water extraction of RR significantly reduced abdominal circumference,INS,low-density lipoprotein-cholesterol(LDL-C),liver lipid accumulation,abdominal adipocyte expansion,PPARγ,TNF-α,IL-1β,GPNMB and NFIA expression in OVX rats,and significantly increased serum E2,miR-29a-3p in the serum exosome and abdominal adipose.Furthermore,Wnt signaling pathway in abdominal adipose was activated by water extraction of RR.CONCLUSION:Water extraction of RR reduced hepatic lipid accumulation through exosomal miR-29a-3p and Wnt signaling pathway in OVX rats.