Tat-modified leptin is more accessible to hypothalamus through brain-blood barrier with a significant inhibition of body-weight gain in high-fat-diet fed mice.
Obesity in human was found mainly due to the poor transportation of leptin through brain-blood barrier (BBB), called as leptin resistance. To produce a leptin capable of penetrating BBB, we have added Tat-PTD(9) to the C terminal of leptin to construct a fusion protein. The fusion Tat-leptin and native leptin genes were synthesized by single-step insertion of a polymerase chain reaction and expressed in Escherichia coli BL21 (Rosseta). The expressing products were purified and renatured by Ni-NTA affinity chromatography, and identified by the molecular size in SDS-PAGE gel and by its immunoreactivity to specific antibody with Western-blotting assay. To bio-functionally evaluate the fusion protein, Balb/c mice fed with high-fat diet (HFD) were given Tat-leptin, leptin or saline for 19 days. The immunohistochemical staining showed the increases in positive stains for the leptin in the region of hypothalamus of the HFD mice with either Tat-leptin or leptin as compared to saline group, but the staining intensity and frequency in the group with Tat-leptin were stronger and higher than those in the group with leptin. Furthermore, the most efficiency in preventing the body-weight gain caused by HFD was found in Tat-leptin group among these three groups. These results suggest that Tat-modified leptin may become a great potential candidate for the prevention or therapy of obese patients.
Zhang
C
C
Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agricultural University, Changchun, China.
Su
Z
Z
Zhao
B
B
Qu
Q
Q
Tan
Y
Y
Cai
L
L
Li
X
X
D000818:Animals; D019440:Anti-Obesity Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000493:pharmacokinetics* / Q000494:pharmacology*; D001812:Blood-Brain Barrier; D004041:Dietary Fats / Q000008:administration & dosage; D016503:Drug Delivery Systems / Q000379:methods; D015756:Genes, tat; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D007150:Immunohistochemistry; D020738:Leptin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000494:pharmacology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009765:Obesity / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000235:genetics / Q000493:pharmacokinetics* / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000494:pharmacology; D013997:Time Factors; D014018:Tissue Distribution; D015430:Weight Gain / Q000187:drug effects; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000493:pharmacokinetics* / Q000494:pharmacology*
D019440:Anti-Obesity Agents; D004041:Dietary Fats; D020738:Leptin; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C549021:Tat-leptin; D054322:tat Gene Products, Human Immunodeficiency Virus; C415751:tat peptide (1-9), Human immunodeficiency virus 1