The Study On Pharmacodynamics And Preparation Of Callicarpa Nudiflora Scald Ointment

Objective:Callicarpa nudiflora Hook.et Arn.is a commonly used medicinal herb in Li nationality of Hainan.There are a lot of pharmacological activities could be found in C.nudiflora,which has been widely using in the clinic.Most of the preparations of C.nudiflora are oral preparations basically used for hemostasis.However,the external use to treat burns and scalds of C.nudiflora has been ignored.In this study,a C.nudiflora external use ointment for burns and scalds be expected to be developed to broaden the range of application the herb.The study was carried out in three stages:the study of the water extraction process of C.nudiflora,the pharmacodynamics and preliminary pharmacological studies of burns and scalds treatment,the preparation of C.nudiflora external use ointment for burns and scalds.Methods:The optimum extraction process was optimized by orthogonal experiment,with the yield of extraction and the extraction rate of acteoside as evaluation indicator.Then the extract of C.nudiflora by this means was used in the second study in which the second-degree scald wounds were induced on dorsal surface of Sprague Dawley adult male rats,then divide them into the following 5 groups:1)control group without treatment,2)positive control group with 1%Silver sulfadiazine(SSD),3)Vaseline group,and groups 4)and 5)that used Vaseline containing 10%and 20%CNE,respectively.These ointments were applied topically once daily in scald wounds for 21 days.To determine the healing effects of CNE,we conducted the gross examination,measurement of wound size,and histopathological examination.To understand the action mechanisms of CNE,the serum levels of tumor necrosis factor alpha(TNF-a),transforming growth factor beta 1(TGF-p1),interleukin-10(IL-10),and vascular endothelial growth factor(VEGF)were measured by enzyme-linked immunosorbent assay.The expression levels of Smad3 in skins were measured by Western blot.Finally,the preparation process was optimized by orthogonal experiment with comprehensive evaluation.Results:The optimized water extract craft of C.nudiflora is:the soak in water for 2 h,the extraction time was 3,with 12,10,10 times water in sequence,and each extraction time was 1-h.This optimized water extract craft is simple and feasible with good repetition.The macroscopic analysis showed CNE accelerated the wound-healing process and the sizes of scald wound were significantly lower on days 15,18,and 21 in the CNE-treated groups compared to the control groups.In particular,the 20%CNE group achieved the highest degree of wound healing.Histopathological evaluation showed the mildly hypertrophic epidermis and the intact dermis in the CNE-treated group,which were apparently distinguished from the control group on day 21.The CNE had no noticeable effect on TNF-a and IL-10 levels on days 2 and 14.The VEGF serum levels of 20%CNE-treated group were significantly(P<0.01)increased on day seven compared to control group.On the 21st day after the scald injury,the TGF-?1 serum levels of 20%CNE-treated group were(P<0.05)lower than the control group.The expression levels of Samd3 were strongly inhibited by 20%CNE,and there was a significant(P<0.05)difference between 20%CNE-treated group and control group in expression levels of Smad3.The optimized preparation process is:First,the aqueous phase(1.2 g tween-80,4.7 g glyceryl monostearate and 0.9 g glycerin)were prepared and fused in the temperature of 70 ?,then slowly added the oleic phase to the oleic phase(10.8 g liquid paraffin,1.4 g span,1.4 g hard paraff-in and 0.9 g vaseline)which had been fused in the temperature of 70 °C by continuous stirring.When the mixture was nearly emulsified adequately,5 g fluid extract of C.nudiflora(including 3 g extract)added slowly into it,emulsifying time is 25 min.After the mix solution cooled down,the C.nudiflora external use ointment for burns and scalds was obtained.This ointment's color was equality,refinement,uniformity,spreadability and stability were good.Conclusions:In conclusion,through the study,obtained the optimized water extract craft of C.nudiflora;,CNE can enhance the wound healing in deep second-degree scald rat model and thus supported its traditional use.These effects may be due to the up-regulation of VEGF in the early stage of wound healing.Also,the CNE can reduce scar formation,which was possibly related to the TGF?1/Smad3 signaling.The obtained C.nudiflora ointment has good appearance and stability,with the potential to become medicines or commodities.