Synergistic properties of garlic and Citrullus colocynthis on reproductive injury caused by diabetes in male rats: Structural and molecular evidence.

This study evaluates the synergistic effect of garlic and Citrullus colocynthis on diabetic reproductive damage by suppressing the AGEs/RAGE/Nox-4 signaling pathway. Thirty-five male Wistar rats were divided into five groups (n = 7/group): Control, Diabetic, Diabetic+G (Garlic, 1 mL/100 g b.w), Diabetic+C (C. colocynthis, 10 mg/kg b.w) and Diabetic+GC (Garlic, 1 mL/100 g b.w and C. colocynthis, 10 mg/kg b.w) groups. At the end of the experimental period (30 days), in diabetic rats, glucose increased, and body & testis weight, luteinizing hormone (LH) and testosterone levels, and sperm count decreased significantly and histopathological injuries were observed. In addition, they have increased testicular apoptosis and oxidative stress. Also, the mechanism based on advanced glycation end products (AGEs)/receptors for advanced glycation end products (RAGE)/NADPH oxidase-4 (Nox-4) was activated in diabetic rats. Separate consumption of garlic and C. colocynthis in Diabetic+G and Diabetic+C groups alleviated the negative adverse effect of diabetes to some extent, but when they were used in the combination form (Diabetic+GC) improvement was profound. Testis histopathology, increased body and testis weight, and enhanced capacity in protecting diabetic reproductive injury was seen. Decreases in testosterone and LH concentration and sperm count in diabetic rats were also reversed by combined administration of garlic and C. colocynthis. It regulated oxidative stress markers, meanwhile reducing caspase-3 immunoexpression. In addition, overexpression of RAGE, Nox-4 and nuclear transcription factor-κB (NF-κB) was inhibited by the combination of garlic and C. colocynthis. PRACTICAL APPLICATIONS: Diabetes mellitus is wide spread all around the world with variety of complications in body including reproductive system in which patients suffer from physical and psychological aspects. Despite many efforts in providing agents for controlling diabetes and its complications, economic conditions of some countries make it difficult for people to provide costly medicine and as a result, they have to bear the complications until they pass away. However, traditional medicine is still finding its way, especially in poor countries with emphasis on medicinal plants. There have been many studies on plants to alleviate diabetes or its side effects. But, using one plant for long term, may be not so effective. Here, we attempted to find whether two plants from two different species can show more efficacy than each one alone. We noticed garlic and Citrullus colocynthis despite having beneficial effects when used alone, they could show synergistic effects in combination.

Isolation of Cysteine-Rich Peptides from Citrullus colocynthis.

The plant Citrullus colocynthis, a member of the squash (Cucurbitaceae) family, has a long history in traditional medicine. Based on the ancient knowledge about the healing properties of herbal preparations, plant-derived small molecules, e.g., salicylic acid, or quinine, have been integral to modern drug discovery. Additionally, many plant families, such as Cucurbitaceae, are known as a rich source for cysteine-rich peptides, which are gaining importance as valuable pharmaceuticals. In this study, we characterized the C. colocynthis peptidome using chemical modification of cysteine residues, and mass shift analysis via matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. We identified the presence of at least 23 cysteine-rich peptides in this plant, and eight novel peptides, named citcol-1 to -8, with a molecular weight between ~3650 and 4160 Da, were purified using reversed-phase high performance liquid chromatography (HPLC), and their amino acid sequences were determined by de novo assignment of b- and y-ion series of proteolytic peptide fragments. In silico analysis of citcol peptides revealed a high sequence similarity to trypsin inhibitor peptides from Cucumis sativus, Momordica cochinchinensis, Momordica macrophylla and Momordica sphaeroidea. Using genome/transcriptome mining it was possible to identify precursor sequences of this peptide family in related Cucurbitaceae species that cluster into trypsin inhibitor and antimicrobial peptides. Based on our analysis, the presence or absence of a crucial Arg/Lys residue at the putative P1 position may be used to classify these common cysteine-rich peptides by functional properties. Despite sequence homology and the common classification into the inhibitor cysteine knot family, these peptides appear to have diverse and additional bioactivities yet to be revealed.