CREG knockdown promotes NIH3T3 Fibroblasts Apoptosis via activating Cathepsin

Background The CREG is an important lysosomal protein involved in a variety of cellular functions including promoting cell differentiation,sustaining mature homeostasis, and antagonizing apoptosis.Deficiency of CREG in cell and tissue results in a pathologic apoptosis.The present study aimed to elucidate the mechanism of CREG regulation apoptosis.Methods We firstly generated stable NIH3T3 fibroblasts by transfection of pDS_shCREGs vectors.Furthermore, PI-Annexin V and TUNEL staining were used to identify that CREG knockdown promoted the cell apoptosis in NIH3T3 fibroblasts.Western blotting and immunofluorescence staining was used to identify the expression and localization of M6P/ IGF2R and cathepsin L in cytoplasm.Results pDS_shCREGs vector transfection produced an approximately 80%decrease in CREG levels both in the lysate and in the media.The expression and localization of M6P/IGF2R and cathepsin L in cytoplasma changed obviously associated with down-regulated of CREG.In addition,the retention and secretion of cathepsin L enhanced significantly.Using the specific inhibitor or siRNA to block cathepsin L activation attenuated the apoptosis mediated by CREG downregulation.Conclusions Our findings indicated that inhibition of CREG expression in NIH3T3 fibroblasts leads to impaired cathepsin L sorting function mediated by M6P/IGF2R and subsequently promotes pathological cell apoptosis.

Influence of phytochemicals in piper betle linn leaf extract on wound healing

Background:Wound healing has being extensively investigated over the world.Healing impairment is caused by many reasons including increasing of free-radicals-mediated damage,delaying in granulation tissue formation,reducing in angiogenesis and decreasing in collagen reorganization.These facts consequently lead to chronic wound healing.Piper betle Linn(Betle)leaves have been folklore used as an ingredient of drugs for cutaneous wound treatment.However,the effect of betle leaf on wound healing is not yet well elucidated.In this study,we aimed to investigate the healing efficacy of methanol leaf extract of Piper betle Linn on proliferation of fibroblast NIH3T3 cells as well as full-thickness burn and excision wounds in swiss mice.Methods:Scratch wound healing assays were conducted to examine the effects of betle leaf extract on healing activity of fibroblast cells.Burn and excision wounds on swiss mouse skins were created for investigating the wound healing progress caused by the betle leaf extract.Malondialdehyde(MDA)was also evaluated to examine the products of lipid hydroperoxide(LPO)under conditions of with or without betle leaf extract treatment.Results:The results of this study showed that Piper betle Linn leaf extract in methanol increased proliferation of NIH3T3 cells and promoted wound healing in vitro and in vivo with both burn wound and excision wound models.In addition,this extract significant decreased level of malondialdehyde(MDA)in liver of treated-mice compared with that in non-treated mice.Conclusions:Our results suggest that Piper betle Linn can be used as an ingredient in developing natural origin drugs for treatment of cutaneous wounds.