Roux-en-Y Gastric Bypass Improves Insulin Sensitivity in Obese Rats with Type 2 Diabetes Mellitus by Regulating the Grin3a/AMPK Signal Axis in Hypothalamic Arcuate Nucleus

Figures

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  Figure 1. RYGB ameliorates insulin resistance and lessens fat accumulation in rats with T2DM. (A) Body weights of rats in each group; (B) The FBG of rats was detected by blood glucose meter; (C) The FSI of rats was tested by kits; (D) HOMA-IR of rats; (E) The changes of plasma TG level of rats were measured by TG kit; (F) The morphological changes of WATs were observed by H&E staining, red arrows represent adipocyte. **P < 0.01.

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  Figure 2. RYGB regulates endoplasmic reticulum stress and neuropeptide levels in arcuate nucleus of T2DM rats. (A–H) qRT-PCR to assess the effects of RYGB on mRNA expression levels of ATF4 (A), IRE-1α (B), PERK (C), CHOP (D), Agrp (E), Npy (F), Pomc (G) and Cart (H) in the arcuate nucleus of hypothalamus in T2DM rats. **P < 0.01.

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  Figure 3. RYGB up-regulates Grin3a expression in white adipose tissue and arcuate nucleus of T2DM rats. (A and B) Western blot to measure the expression levels of Grin3a protein in white adipose tissue (A) and arcuate nucleus tissue (B) of rats in each group; (C) qRT-PCR to test the mRNA expression level of Grin3a in arcuate nucleus of rats. **P < 0.01.

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  Figure 4. RYGB activates AMPK signals in the arcuate nucleus of T2DM rats. The protein expression levels of p-AMPK and AMPK in the arcuate nucleus of rats in each group were detected by Western blot. **P < 0.01.

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  Figure 5. Knocking down Grin3a promotes lipid accumulation and production in 3T3-L1 cells. (A) The expression level of Grin3a in each group was detected by qRT-PCR; (B) TG level in cells of each group was checked by TG kit; (C) the formation of lipid droplets in 3T3-L1 cells was observed by oil red O staining; (D–G) The mRNA expression levels of PPARγ (D), C/ebpα (E), FASN (F) and FABP4 (G) were determined by qRT-PCR. *P < 0.05, **P < 0.01.

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  Figure 6. Knocking down Grin3a reverses the role of RYGB in reducing insulin resistance and fat accumulation in T2DM rats. (A) The expression level of Grin3a in arcuate nucleus of rats was detected by qRT-PCR; (B) Body weight of rats; (C) The FBG levels of rats were measured by blood glucose meter; (D) The FSI levels of rats in each group were checked by kits; (E) HOMA-IR of rats; (F) TG kit to test the level of TG in plasma of rats; (G) The WAT of rats in each group was observed by H&E staining, red arrows represent adipocyte. **P < 0.01.

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  Figure 7. Knocking down Grin3a reverses the impact of RYGB on endoplasmic reticulum stress and neuropeptide levels in arcuate nucleus of T2DM rats. The mRNA expression levels of ATF4 (A), IRE-1α (B), PERK (C), CHOP (D), Agrp (E), Npy (F), Pomc (G) and Cart (H) in arcuate nucleus tissue of rats in each group were measured by qRT-PCR. **P < 0.01.

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  Figure 8. RYGB improves insulin resistance in T2DM rats by regulating Grin3a/AMPK signal axis. Western blot was employed to measure the protein expression changes of Grin3a, AMPK and p-AMPK in rats of each group. **P < 0.01.