Peptide bifonctionnel
A bifunctional peptide and sequence technology, applied in the direction of peptides, specific peptides, fusion polypeptides, etc., can solve the problem of not reducing the expression of enzymes
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Embodiment 1
[0116] Example 1: Effects of Bifunctional Peptides on Collagen Synthesis-Protein Analysis
[0117] Dermal fibroblasts were isolated from abdominal skin biopsies obtained during surgical procedures in healthy patients between the ages of 35 and 75.
[0118] After removing the epidermis, fibroblasts were recovered by enzymatic digestion, followed by culture at 37°C in DMEM (Dulbecco's Modified Eagle's Medium) supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin .
[0119] Next, the fibroblasts were treated with a peptide having the S1 sequence or fragments thereof at a concentration of 100 μg / ml. The various fragments tested are as follows:
[0120] - Peptide 1 (pept1): peptide with S1 sequence
[0121] - Peptide 2 (pept2): Sequence N-(Val-Gly-Val-Ala-Pro-Gly) 3 Arg-Val-Arg-Leu-OH
[0122] - Peptide 3 (pept3): N-(Val-Gly-Val-Ala-Pro-Gly) 3 -OH
[0123] - Peptide 4 (pept4): N-Arg-Val-Arg-Leu-Gly-Ile-Leu-OH
[0124] - Peptide 5 (pept5): N-Gly-Ile-Leu-O...
Embodiment 2
[0130] Example 2 : Effect of Bifunctional Peptides on Collagen Synthesis - Gene Expression Analysis
[0131]Gene expression of collagen type I and type III in fibroblast cultures was tested in the presence of different peptides (pept1 to pept5) at a concentration of 100 μg / ml.
[0132] After 24 hours of treatment with different peptides, using Kit (QIAGEN) RNAs were extracted from the culture, followed by the use from SABiosciences TM The RT2 First Strand Kit (RT2First Strand Kit) retranscribes RNAs into complementary DNAs (cDNAs).
[0133] Next, real-time PCR analysis was performed in a 96-well plate using a Mx3000p thermal cycler from Stratagene. Each sample was analyzed in triplicate. cDNAs containing RT2 Green / ROX TM qPCR Master Mix reagents (SABiosciences TM ) and sense and antisense primers specific for the gene to be amplified are used as substrates in the reaction mixture. Fluorescence was read at the end of each cycle and the readings were analyzed by MxPr...
Embodiment 3
[0137] Example 3 : Effect of Bifunctional Peptides on Collagen Synthesis - Protein and Gene Expression Tests According to Patient Age
[0138] Dermal fibroblasts from younger or older patients (35, 42 and 53 years old) were cultured and the peptides of the invention having the S1 sequence were tested for protein and gene expression of type I and type III collagen. Figure 5 The results obtained for the protein expression of the two types of collagen can be seen. It can be seen that the effect of the peptides of the present invention on the synthesis of collagen is greater when the fibroblasts are from older patients.
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