Abstract
Backround: Erectile dysfunction (ED) is the inability of male reproductive organs within sexual intercourse caused by neurogenic and hormonal disorders. There are some causes of ED such as hypertension, stress, neurological disorders, stroke, diabetes, atherosclerosis, lifestyle, alcohol, smoking, and age-related hormonal decline can cause infertility. The natural treatment of sexual dysfunction through aphrodisiac activity of the plant is to increase sexual hormones, spermatogenesis activity and through PDE5 inhibitors such as sildenafil, verdenafil, and tadalafil which can inhibit the hydrolysis of second messenger cGMP of penis smooth muscle cells. Primer Design for amplification of several PDE5 gene nucleotide sequences obtained from NCBI gen banks and tested directly through explosions at NCBI and also using MEGA 6, primer 3 plus, and fast PCR software. The natural treatment of sexual dysfunction through aphrodisiac activity from plants to increase sexual hormone, spermatogenesis activity and inhibitor PDE5 such as sildenafil, vardenafil, and tadalafil can inhibit hydrolysis second messenger of cells cGMP smooth penis muscles. Method: Primer design stages for several sequences are data supply, multiple sequence alignment, sequence trimming, primer design (fastPCR input), in silico PCR analysis, and primer evaluation (Primer Test, OligoCalc and BLAST). Results: Primer of PDE5 that is choosen is with reverse sequence 5-TGCATTGACCATGTCTCTCGTT-3, forward 5-CGCCGATCTGGGCTGAACTA-3 able to amplify template DNA at temperature 67.2°C, 65.8°C, 63.7°C, however, the DNA band fragment looks not very clear, while it is more clearly seen at Tm temperature 61.2°C, 59.1°C, 57.8°C and 57°C. Conclusion: PDE5 primers can be amplified well at temperature 61.2°C, 59.1°C, 57.8°C and 57°C. PDE5 primer succeeded in amplifying DNA with a product length of 402 bp.
Highlights
Phospodiesterase is a group of enzymes that break down substrates such as phosphodiaterases which bind pyrophosphates from nucleotides and nucleotide sugars [1]
The natural treatment of sexual dysfunction through aphrodisiac activity of the plant is to increase sexual hormones, spermatogenesis activity and through PDE5 inhibitors such as sildenafil, verdenafil, and tadalafil which can inhibit the hydrolysis of second messenger cyclic GMP (cGMP) of penis smooth muscle cells
PDE5 is mediated by cyclic AMP (cAMP) and cGMP so it is used as a therapy to treat erectile dysfunction, asthma, heart failure, inflammation and depression [5]
Summary
Phospodiesterase is a group of enzymes that break down substrates such as phosphodiaterases which bind pyrophosphates from nucleotides and nucleotide sugars [1]. PDE5 degrades cyclic AMP (cAMP) and cyclic GMP (cGMP) second messenger molecules [2]. PDE5 has important role in transferring signal transduction because PDE5 regulates nucleotide signals [3]. PDE5 often becomes the target of drugs, because it has the ability to regulate small levels of nucleotides, tissue distribution, substrate specificity and parmocology [4]. PDE5 is mediated by cAMP and cGMP so it is used as a therapy to treat erectile dysfunction, asthma, heart failure, inflammation and depression [5]. PDE5 plays a role in cAMP and cGMP degradation. The role of PDE5 is related to adenylate silage (AC) and guanylate cyclase (GC) in regulation and the duration of the intracellular signaling mechanism [6]. CGMP hydrolyzes PDE5 which is expressed in vascular smooth muscle The role of PDE5 is related to adenylate silage (AC) and guanylate cyclase (GC) in regulation and the duration of the intracellular signaling mechanism [6]. cGMP hydrolyzes PDE5 which is expressed in vascular smooth muscle
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