Metabolic Consequences of Neuronal HIF1α-Deficiency in Mediobasal Hypothalamus in Mice.

Objective: This study aims to investigate whether hypoxia-inducible factor 1α (HIF1α) in the neurons of the mediobasal hypothalamus is involved in the regulation of body weight, glucose, and lipid metabolism in mice and to explore the underlying molecular mechanisms. Methods: HIF1α flox/flox mice were used. The adeno-associated virus that contained either cre, GFP and syn, or GFP and syn (controls) was injected into the mediobasal hypothalamus to selectively knock out HIF1α in the neurons of the mediobasal hypothalamus. The body weight and food intake were weighed daily. The levels of blood glucose, insulin, total cholesterol (TC), triglyceride (TG), free fatty acid (FFA), high-density lipoprotein (HDL), and low-density lipoprotein (LDL)were tested. Intraperitoneal glucose tolerance test (IPGTT) was performed. The insulin-stimulated Akt phosphorylation in the liver, epididymal fat, and skeletal muscle were examined. Also, the mRNA expression levels of HIF1α, proopiomelanocortin (POMC), neuropeptide Y (NPY), and glucose transporter protein 4 (Glut4) in the hypothalamus were checked. Results: After selectively knocking out HIF1α in the neurons of the mediobasal hypothalamus (HIF1αKOMBH), the body weights and food intake of mice increased significantly compared with the control mice (p < 0.001 at 4 weeks). Compared with that of the control group, the insulin level of HIF1αKOMBH mice was 3.5 times higher (p < 0.01). The results of the IPGTT showed that the blood glucose level of the HIF1αKOMBH group at 20-120 min was significantly higher than that of the control group (p < 0.05). The serum TC, FFA, HDL, and LDL content of the HIF1αKOMBH group was significantly higher than those of the control group (p < 0.05). Western blot results showed that compared with those in the control group, insulin-induced AKT phosphorylation levels in liver, epididymal fat, and skeletal muscle in the HIF1αKOMBH group were not as significantly elevated as in the control group. Reverse transcription-polymerase chain reaction (RT-PCR) results in the whole hypothalamus showed a significant decrease in Glut4 mRNA expression. And the mRNA expression levels of HIF1α, POMC, and NPY of the HIF1αKOMBH group decreased significantly in ventral hypothalamus. Conclusions: The hypothalamic neuronal HIF1α plays an important role in the regulation of body weight balance in mice under normoxic condition. In the absence of hypothalamic neuronal HIF1α, the mice gained weight with increased appetite, accompanied with abnormal glucose and lipid metabolism. POMC and Glut4 may be responsible for this effect of HIF1α.

Intermittent hypoxia induces tumor immune escape in murine S180 solid tumors via the upregulation of TGF-ß1 in mice.

PURPOSE: Studies have shown that intermittent hypoxia (IH) alters host immune functions and promotes tumor growth. However, the relevant mechanisms of these effects have not been completely elucidated. We hypothesized that IH promotes the growth of tumors by changing cytokine levels in the tumor microenvironment and inducing immune escape. METHODS: Sarcoma-180 (S180) solid tumor cells were injected into the right flank of Kunming mice. The mice were then randomly divided into the IH and room air (RA) groups. The mice were euthanized 2 weeks after IH exposure, and the weight of tumor tissues was measured. Next, IL-6, IL-17, IL-10, and TNF-α levels in tumor tissues were measured via enzyme linked immunosorbent assay (ELISA), and hypoxia inducible factor-1α (HIF-1α) and transforming growth factor ß1 (TGF-ß1) expressions were examined through Western blot analysis. RESULTS: Two weeks of IH exposure significantly accelerated the growth of S180 solid tumors. Western blot analysis results showed that the expression levels of HIF-1α and TGF-ß1 in S180 tumors in the IH group were significantly upregulated compared with those in the RA group. ELISA results showed that compared with the RA group, the IH group had significantly increased TNF-α and IL-10 (P < 0.05) and significantly decreased IL-17 (P < 0.05). CONCLUSION: IH might promote the growth of S180 solid tumors by inhibiting the antitumor immune response and inducing tumor immune escape via the upregulation of TGF-ß1.

Testicular AQP1 expression in a rat model of testicular Ischemia-Reperfusion injury.

Aquaporin 1 (AQP1) is the archetype of all aquaporins and involved in rapid cellular water fluxes and cell volume regulation. AN OBJECTIVE: This study was conducted for the investigation of AQP1 expression in normal testicular tissues and those with I/R injury in a rat model. STUDY DESIGN: A TT rat model was established using male Wister rats (4 weeks old, 180-220 g), and AQP1 distribution in the testicular tissues was detected by immunohistochemistry. The wet/dry (W/D) weight ratios of the testes were determined at 12 h, 24 h, 36 h, 48 h, or 5 days after the establishment of the TT model. At each time point, pathological sections were prepared and the mRNA and protein expression levels of AQP1 were determined by RT-qPCR and Western blotting, respectively. RESULTS: Immunohistochemical staining indicated that AQP1 distributes in testicular vascular endothelial cells and interstitial connective tissues. The testicular edema was observed 12 and 24 h after TT, as indicated by the increase in wet/dry weight ratio and pathological changes, such as enlarged testicular interstitium, atrophy of spermatogenic tubules, and epineurium tubule exfoliation. Increase in the expression levels of Aqp1 mRNA and AQP1 protein levels peaked at 24 h. Edema was alleviated at 36 and 48 h, as manifested by the gradual thinning of the spermatogenic tubules epithelium with narrowed interstitium and weakened inflammatory cell infiltration. Meanwhile, the mRNA and protein levels of AQP1 dramatically decreased. At 5 days after TT, edema was nearly absent, and the mRNA and protein levels of AQP1 were restored to basal levels. DISCUSSION: Testicular torsion increases AQP1 expression and W/D ratios in testis tissues. The upregulation of AQP1 expression and decline in AQP1 level are consistent to the development and alleviation of edema in testis tissues that underwent testicular torsion. CONCLUSION: Changes in AQP1 expression were consistent with edema severity in the testes, indicating a close relationship between the expression of AQP1 and the extent of edema in testicular I/R.