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. 2015 Apr 30;13(5):2732-56.
doi: 10.3390/md13052732.

Anti-obese effect of glucosamine and chitosan oligosaccharide in high-fat diet-induced obese rats

Lanlan Huang  1 , Jian Chen  2 , Peiqiu Cao  3 , Haitao Pan  4 , Chen Ding  5 , Tiancun Xiao  6   7 , Pengfei Zhang  7 , Jiao Guo  8 , Zhengquan Su  9
Affiliations

Anti-obese effect of glucosamine and chitosan oligosaccharide in high-fat diet-induced obese rats

Lanlan Huang et al. Mar Drugs. .

Abstract

Objective: This study is to evaluate the anti-obese effects of glucosamine (GLC) and chitosan oligosaccharide (COS) on high-fat diet-induced obese rats.

Methods: The rats were randomly divided into twelve groups: a normal diet group (NF), a high-fat diet group (HF), Orlistat group, GLC high-, middle-, and low-dose groups (GLC-H, GLC-M, GLC-L), COS1 (COS, number-average molecular weight ≤1000) high-, middle-, and low-dose groups (COS1-H, COS1-M, COS1-L), and COS2 (COS, number-average molecular weight ≤3000) high-, middle-, and low-dose groups (COS2-H, COS2-M, COS2-L). All groups received oral treatment by gavage once daily for a period of six weeks.

Results: Rats fed with COS1 gained the least weight among all the groups (P < 0.01), and these rats lost more weight than those treated with Orlistat. In addition to the COS2-H and Orlistat groups, the serum total cholesterol (CHO) and low-density lipoprotein cholesterol (LDL-C) levels were significantly reduced in all treatment groups compared to the HF group (P < 0.01). The various doses of GLC, COS1 and COS2 reduced the expression levels of PPARγ and LXRα mRNA in the white adipose tissue.

Conclusions: The results above demonstrated that GLC, COS1, and COS2 improved dyslipidemia and prevented body weight gains by inhibiting the adipocyte differentiation in obese rats induced by a high-fat diet. Thus, these agents may potentially be used to treat obesity.

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Figures

Figure 1
Figure 1
The effect of GLC (A), COS1 and COS2 (B) to rats’ weekly diet intake during administration (n = 10, means ± SD). Food intake of rats gradually increased for four weeks and then declined during the following two weeks. In Figure 1B, the food intake of rats was decreased in the COS1-H, COS1-M, COS1-L, COS2-M and COS2-L groups when compared with Orlistat. Abbreviations: GLC, glucosamine; COS, chitosan oligosaccharide; COS1, number-average molecular weight ≤1000; COS2, number-average molecular weight ≤3000; NF, normal diet group; HF, high-fat diet group; GLC-H, GLC-M, GLC-L, GLC high-, middle-, and low-dose groups; COS1-H, COS1-M, COS1-L, COS1 high-, middle-, and low-dose groups; COS2-H, COS2-M, COS2-L, COS2 high-, middle-, and low-dose groups.
Figure 2
Figure 2
The effect of GLC (A), COS1 and COS2 (B) to rats’ weight (n = 10, means ± SD). After modeling, the weight of all rats increased, but NF group had lower gain, which significantly differed from those of the other groups. The treatment groups tended to gain weight slower than the HF group, which indicated that GLC, COS1 and COS2 could inhibit weight gain in rats.
Figure 3
Figure 3
The effect of GLC, COS1 and COS2 on fat pat (A) and body fat (B) ratio in rats (n = 10, means ± SD). Orlistat, GLC-H, and COS2-H (P < 0.05) could reduce the body fat and body fat ratio in obese rats when compared with the HF group, which carried high fat. Besides, it appeared dose dependent in the COS2 group. Note: Compared with HF, * P < 0.05; ** P < 0.01; Compared with Orlistat, # P < 0.05; ## P < 0.01.
Figure 4
Figure 4
The effect of GLC, COS1 and COS2 on serum CHO, TG, HDL-C and LDL-C in rats (n = 10, means ± SD). (A) GLC, COS1 and COS2 could reduce the CHO in the serum, which were better than Orlistat; (B) Compared with the HF group, the TG in the serum was decreased in GLC, COS1 and COS2; (C) COS2-H and COS2-M significantly increased the HDL-C in the serum, and it appeared dose dependent in the COS1 group; (D) GLC, COS1 and COS2 reduced the LDL-C in the serum more effectively than Orlistat. Note: Compared with HF, * P < 0.05; ** P < 0.01; Compared with Orlistat, # P < 0.05; ## P < 0.01.
Figure 5
Figure 5
The effect of GLC, COS1 and COS2 on liver morphologies in the rats. The livers of rats in the NF group were small, bright red, smooth and had sharp edges. In contrast, those in the HF group were light yellow, swelling, thick edge and had plenty of visible fat granules. However, this state of fatty liver was relieved in treatment groups, implying that GLC, COS1 and COS2 could reduce fat deposition in the liver, and protect the liver.
Figure 6
Figure 6
Histopathology of liver in rats. The liver tissues in the HF group developed a high degree of steatosis, hepatocytes with severe fat vacuoles and the infiltration of inflammatory cells. The symptoms of fatty liver were mitigated in varying degrees in Orlistat, GLC, COS1 and COS2 groups.
Figure 7
Figure 7
Histopathology of heart in rats. In the NF group, myocardial cells arranged orderly and cardiac muscle fibers showed clear. The heart sections of HF rats showed disarrayed myocardial fibers, interstitial fibrosis and the infiltration of inflammatory cells. Then, this state of rhabdomyolysis were alleviated in treatment groups.
Figure 8
Figure 8
Histopathology of mesenteric fatty and subcutaneous fatty tissue. The adipocytes of the HF group were larger than those of other groups. Treatment with Orlistat, GLC, COS1 and COS2 clearly inhibited the proliferation of adipocytes. Note: (A) mesenteric fatty tissue; (B) subcutaneous fatty tissue.
Figure 9
Figure 9
The Statistics of differentially expressed genes. Compared with the HF group, five hundred nineteen genes were up-regulated, while 127 were down-regulated in the GLC group. When COS1 group compared with the HF group, there were six hundred forty genes up-regulated and three hundred twenty five genes down-regulated.
Figure 10
Figure 10
PPAR signaling pathway. (A) HF vs. NF PPAR Signaling Pathway; (B) COS1 vs. NF PPAR Signaling Pathway. ↑ Gene was up-regulated; ↓ Gene was down-regulated.
Figure 11
Figure 11
The mRNA expression levels of epididymal fat PPARγ (A) and LXRα (B) in the different groups. The level of mRNA was detected by Q-PCR. ΔCt is the average value of 10 samples in the formulation (average mRNA expression of experiment groups/average mRNA expression of NF) = 2−∆∆Ct = 2−(∆Ct control − ∆Ct FF). If 2−∆∆Ct < 1, the average mRNA expression of the experiment groups is lower than that in NF. If 2−∆∆Ct > 1, the average mRNA expression of the experiment groups is higher than that in NF. Note: Compared with HF, * P < 0.05; ** P < 0.01; Compared with NF, # P < 0.05; ## P< 0.01.
Figure 12
Figure 12
PPARγ and LXRα mRNA in adipose tissue. PPARγ and LXRα function as obligate heterodimers with retinoid X receptors (RXR) and influence gene transcription via multiple mechanisms. In keeping with their functions as lipid sensors, ligand-bound PPAR or LXR activate feed-forward metabolic cascades that regulate lipid homeostasis via the transcription of genes involved in lipid metabolism, storage, and transport. PPARγ activates a battery of genes involved in lipid storage and lipogenesis, allowing adipose tissue safely to store greater quantities of fat and undergo differentiation.
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