Effect of curcumin analogue H8 on visceral fat metabolism in db/db mice
-
摘要:
目的 探讨姜黄素类似物H8对db/db小鼠(糖尿病模型)内脏脂肪代谢的影响及机制。 方法 将db/db小鼠随机分为模型组、阳性对照组(罗格列酮5 mg/kg),H8低、高剂量组(5、10 mg/kg);连续干预8周,称量小鼠体重和内脏脂肪重量,检测血液生化指标,取肾周脂肪组织石蜡包埋,HE染色观察内脏脂肪细胞,免疫荧光法检测内脏脂肪细胞F4/80表达水平;Real-time PCR法检测脂代谢合成酶(FAS)、乙酰辅酶A羧化酶1(ACC1)、固醇调节元件绑定蛋白(SREBP)mRNA表达水平,蛋白印记法检测FAS、ACC1、SREBP的蛋白表达。 结果 与模型组比较,低、高剂量H8组小鼠内脏脂肪系数[分别为(3.20 ± 0.23)%、(2.93 ± 0.75)%]明显降低、内脏脂肪细胞体积[分别为(304.7 ± 24.8)、(331.3 ± 18.0)明显缩小(均P < 0.01),内脏脂肪细胞F4/80表达水平[分别为(0.39 ± 0.06)、(0.43 ± 0.04)]显著降低,脂肪合成相关基因FAS[(217.0 ± 46.5)、(249.1 ± 31.4)]、ACC1[(54.9 ± 5.4)、(50.4 ± 5.3)]、SREBP[(14.2 ± 1.7)、(13.4 ± 1.5)] mRNA表达水平均明显下降(均P < 0.01),脂肪合成相关蛋白FAS[(0.69 ± 0.03)、(0.41 ± 0.06)]、ACC1[(0.11 ± 0.02)、(0.07 ± 0.01)]、SREBP[(0.36 ± 0.05)、(0.41 ± 0.06)]表达也显著降低(均P < 0.01)。 结论 姜黄素类似物H8能够改善db/db小鼠内脏脂肪代谢。 Abstract:Objective To investigate the effect and mechanism of curcumin analogue H8 on visceral fat metabolism in db/db mice (diabetes model). Methods Totally 32 specific pathogen free 8-week db/db mice were randomly divided into a model group (with sodium carboxymethyl cellulose), a positive control group (with 5 mg/kg rosiglitone), and low and high dose (5, 10 mg/kg) curcumin H8 groups. The administrations were performed with gastric gavage once a day continuously for 8 weeks. At the end of the experiment, the body and visceral fat weight of the mice were weighed; the blood biochemical indexes were detected; the perirenal fat tissue was embedded in paraffin; the visceral fat cells were observed with hematoxylin eosin staining; and the expression level of F4/80 in visceral fat cells was detected with immunofluorescence. Real-time PCR was used to detect the expressions of lipid metabolism synthase (FAS), acetyl-CoA carboxylase 1 (ACC1), and sterol regulatory element binding protein (SREBP) mRNA; the protein expression of FAS, ACC1 and SREBP were detected by Western blot. Results Compared with those in the model group, all indicators of visceral fat metabolism decreased significantly in the low and high dose curcumin H8 group (P < 0.01 for all); the indicators included visceral fat coefficient (3.20 ± 0.23% and 2.93 ± 0.75%), volume of visceral fat cells (304.7 ± 24.8 and 331.3 ± 18.0), F4/80 in visceral fat cells (0.39 ± 0.06 and 0.43 ± 0.04), FAS mRNA (217.0 ± 46.5 and 249.1 ± 31.4), ACC1mRNA (54.9 ± 5.4 and 50.4 ± 5.3), SREBP mRNA (14.2 ± 1.7 and 13.4 ± 1.5), FAS protein (0.69 ± 0.03 and 0.41 ± 0.06), ACC1 protein (0.11 ± 0.02 and 0.07 ± 0.01), and SREBP protein (0.36 ± 0.05 and 0.41 ± 0.06). Conclusion Curcumin analog H8 can improve visceral fat metabolism in db/db mice. -
Key words:
- diabetes mellitus /
- lipid metabolism disorder /
- fatty acid synthetase
-
表 1 脂肪合成相关基因mRNA引物序列
基因 上游引物(5′ – 3′) 下游引物(5′ – 3′) 片段长度(bp) FAS NM_007988.3 GTTGGCCCAGAACTCCTGTA GTCGTCTGCCTCCAGAGC 110 ACC1 NM_133360.2 GGAGATGTACGCTGACCGAGAA ACCCGACGCATGGTTTTCA 105 SREBP NM_011480.4 TGGTTGTTGATGAGCTGGAG GGCTCTGGAACAGACACTGG 90 RPS16 NM_013647.2 CGTGCTTGTGCTCGGAGCTA GCTCCTTGCCCAGAAGCAAA 203 表 2 H8对db/db小鼠内脏脂肪重量影响(
$\bar x$ ± s,n = 8)组别 体重(g) 脂肪重量(g) 脂肪系数 模型组 59.75 ± 1.567 3.23 ± 0.22 5.7 ± 0.20 罗格列酮组 49.01 ± 2.858a 2.63 ± 0.15a 3.96 ± 0.24a H8 低剂量组 48.89 ± 3.766a 1.55 ± 0.2b 3.20 ± 0.23b H8 高剂量组 46.87 ± 2.024a 1.38 ± 0.37b 2.93 ± 0.75b 注:与模型组比较,a P < 0.05,b P < 0.01。 表 3 H8对db/db小鼠血液生化指标影响(mg/dL,
$\bar x$ ± s,n = 8)组别 脂蛋白a 胆固醇 甘油三酯 HDL LDL 血糖 模型组 0.671 ± 0.06 169.2 ± 10.5 108.6 ± 7.71 113.2 ± 6.5 19.7 ± 1.2 443.3 ± 31.2 罗格列酮组 0.621 ± 0.03 99.4 ± 1.3 b 74.5 ± 2.21 b 88.8 ± 2.7 10.7 ± 0.2 b 169.4 ± 13.2 b H8低剂量组 0.430 ± 0.02 a 132.5 ± 6.2 81.3 ± 4.1 a 123.6 ± 3.4 c 14.2 ± 0.7 b 230.1 ± 18.6 b H8高剂量组 0.300 ± 0.04 bc 130.1 ± 9.6 77.4 ± 3.5 b 134.1 ± 7.4 ac 13.4 ± 1.2 b 194.3 ± 21.4 b 注:与模型组比较,a P < 0.05,b P < 0.01;与罗格列酮组比较,c P < 0.01。 表 4 H8对db/db小鼠脂肪组织中脂肪合成相关基因表达影响(
$\bar x$ ± s,n = 8)组别 FAS ACC1 SREBP 模型组 511.0 ± 32.5 78.1 ± 19.0 25.6 ± 3.9 罗格列酮组 204.9 ± 35.3 b 57.6 ± 25.7 a 15.1 ± 3.2 a H8低剂量组 217.0 ± 46.5 b 54.9 ± 5.4 b 14.2 ± 1.7 b H8高剂量组 249.1 ± 31.4 b 50.4 ± 5.3 b 13.4 ± 1.5 b 注:与模型组比较,a P < 0.05,b P < 0.01。 表 5 H8对db/db小鼠脂肪组织中脂肪合成相关蛋白表达影响(
$\bar x$ ± s,n = 8)组别 FAS ACC1 SREBP 模型组 1.08 ± 0.08 0.34 ± 0.05 1.24 ± 0.13 罗格列酮组 0.71 ± 0.01 a 0.18 ± 0.03 a 0.82. ± 0.08 a H8低剂量组 0.69 ± 0.03 b 0.11 ± 0.02 b 0.36 ± 0.05 b H8高剂量组 0.41 ± 0.06 b 0.07 ± 0.01 b 0.41 ± 0.06 b 注:与模型组比较,a P < 0.05,b P < 0.01。 -
[1] Chen QY, Zheng Y, Jiao DM, et al. Curcumin inhibits lung cancer cell migration and invasion through Rac1-dependent signaling pathway[J]. J Nutr Biochem, 2014, 25(2): 177 – 185. doi: 10.1016/j.jnutbio.2013.10.004 [2] He Y, Yue Y, Zheng X, et al. Curcumin, inflammation, and chronic diseases: how are they linked?[J]. Molecules, 2015, 20(5): 9183 – 9213. doi: 10.3390/molecules20059183 [3] Naksuriya O, Okonogi S, Schiffelers RM, et al. Curcumin nanoformulations: a review of pharmaceutical properties and preclinical studies and clinical data related to cancer treatment[J]. Biomaterials, 2014, 35(10): 3365 – 3383. doi: 10.1016/j.biomaterials.2013.12.090 [4] 秦培洁, 张栋伟, 莫芳芳, 等. 姜黄素对胰岛素抵抗3T3-L1前脂肪细胞糖脂代谢及细胞分化的影响[J]. 北京中医药大学学报, 2014, 37(4): 236 – 239. doi: 10.3969/j.issn.1006-2157.2014.04.005 [5] Aggarwal BB, Gupta SC, Sung B, et al. Curcumin: an orally bioavailable blocker of TNF and other pro-inflammatory biomarkers[J]. Br J Pharmacol, 2013, 169(8): 1672 – 1692. doi: 10.1111/bph.12131 [6] Yuan X, Li H, Bai H, et al. The 11β-hydroxysteroid dehydrogenase type 1 inhibitor protects against the insulin resistance and hepatic steatosis in db/db mice[J]. Eur J Pharmacol, 2016, 788: 140 – 151. doi: 10.1016/j.ejphar.2016.05.034 [7] Yuan XH, Li H, Bai H, et al. Synthesis of novel curcumin analogues for Inhibition of 11β-hydroxysteroid dehydrogenase type 1 with anti-diabetic properties[J]. Eur J Med Chem, 2014, 77: 223 – 230. doi: 10.1016/j.ejmech.2014.03.012 [8] 于冬青, 邓华聪. 姜黄素对糖尿病大鼠糖、脂代谢及氧化应激的影响[J]. 重庆医学, 2015, 34(1): 37 – 39. [9] Bettaieb A, Cremonini E, Kang H, et al. Anti-inflammatory actions of (-)-epicatechin in the adipose tissue of obese mice[J]. Int J Biochem Cell Biol, 2016, 81(Pt B): 383 – 392. [10] Masi LN, Martins AR, Crisma AR, et al. Combination of a high-fat diet with sweetened condensed milk exacerbates inflammation and insulin resistance induced by each separately in mice[J]. Sci Rep, 2017, 7(1): 3937 – 3947. doi: 10.1038/s41598-017-04308-1 [11] Alsanea S, Liu D. BITC and S-carvone restrain high-fat diet-induced obesity and ameliorate hepatic steatosis and insulin resistance[J]. Pharm Res, 2017, 34(11): 2241 – 2249. doi: 10.1007/s11095-017-2230-3 [12] 占敏霞, 巫冠中. 脂肪酸合成酶与疾病[J]. 亚太传统医药, 2012, 8(9): 210 – 212. doi: 10.3969/j.issn.1673-2197.2012.09.117 [13] Palou M, Priego T, Sánchez J, et al. Gene expression patterns in visceral and subcutaneous adipose depots in rats are linked to their morphologic features[J]. Cell Physiol Biochem, 2009, 24(5-6): 547 – 556. doi: 10.1159/000257511 [14] Oberkofler H, Fukushima N, Esterbauer H, et al. Sterol regulatory element binding proteins: relationship of adipose tissue gene expression with obesity in humans[J]. Biochim Biophys Acta, 2002, 1575(1-3): 75 – 81. doi: 10.1016/S0167-4781(02)00279-8