Diabetes, Chronic Skin Ulcers Yin Syndrome Model And Huiyang Ointment Diabetic Ulcers Yin Syndrome Impact At Different Phases

ObjectiveTo establish the rat model of diatbtic skin ulcers and observe the influences of HuiyangShengji ointment and its modified formulae on chronic skin ulcer of diabetic rats at different phase of inflammation factor and related effects of promoting healing factor mRNA expression.To explore the mechanism of action of Huiyang Shengji ointment on the healing of chronic skin ulcer.Methods1Male SD rats (n=50) were randomly divided into five groups,10rats in each group:skin defect group (group QS:dermal deficiency),diabetic group (group DM:STZ injection+skin excision),diabetes plus Staphylococcus aurous group(group DMJJ:STZ injection+skin excision+bacterial infection),diabetes plus hydrocortisone group (group DMJS:STZ injection+skin excision+hydrocortisone intervention) and diabetes plus hydrocortisone and implantation of foreign body group (DMYW group:injection STZ+skin excision+hydrocortisone intervention+foreign body embedded).The general condition and the character of wound surface were observed and wound area was recorded and wound healing rate was calculated.Compared the different groups’ wound area and the organic morphology of the granulation tissue stained by hematoxylin and eosin (H&E) were observed.2Male SD160rats were randomly selected6as a blank control group (blank group),without handling.The remaining rats were randomly divided into five groups after modeling diabetes2w:1d,3d,5d,7d,14d group.Then,these group divided into normal control group(normal group:Vaseline ointments),model control group (model group: Vaseline ointments), Huiyang Shengji ointment group (whole formula group:Huiyang Shengji ointment whole formula ointments), Wenyang Yiqi group (Yiqi Group:group of modified Wenyang Yiqi formulaointments)and HuoXueShengJi group (Huoxue group:group of modified Huoxue Shengji formula ointments). Normal group using the method of skin excision, other groups using " STZ injection-hydrocortisone interference-skin excision-foreign body embedded" preparation of composite factors for chronic skin ulcer model. After the appropriate treatment period, executing the rats and testing the content of IL-1α and TXB2in serum by enzyme-linked immunosorbent assay(ELISA) of five time points.3By enzyme-linked immunosorbent assay(ELISA) and using RT-PCR assay for detection of treatment3d,7d,14d, eNOS expression in the serum and granulation tissue.Results 1The " STZ injection-hydrocortisone interference-skin excision-foreign body embedded " composite factor model with other modeling method, can significantly prolong the time of wound healing. The model rats showed the basic features of syndrome of yang deficiency, such as loose bowels, eating less, weight loss, cold tail drowsiness and activity decrease, eg. The HE staining showed that The new skin tissue in rats of group QS structure, clear hierarchy, cuticle distribution of new skin, collagen fiber cell thick irregular distribution, the dermis layer of new size distribution of blood vessels; Group DM with single epidermal cell generation, has a fine collagen fiber and a large number of inflammatory cells within the dermis, no neovascularization; in group DMJJ, multilayer neonatal epidermal cells regular distribution, organization within the tissue fluid, plasma coagulation components and infiltration of inflammatory cells, a small amount of angiogenesis; group DMJS edge of the dermis without epidermal tissue generation, a small amount of inflammatory cell infiltration and tissue fluid, no angiogenesis; the DMYW group produced in the epidermis, no infiltration of inflammatory cells, without healing tendency. The MASSON staining showed that under blue light showed collagen fibers, QS group of collagen fiber thick long, dense and regular arrangement; DM group of collagen fiber is coarse, but less and fine than the QS group, the inflammatory cells arranged in disorde; DMJJ group of collagen fiber short, scattered on the intercellular; DMJS group of collagen fiber fine, and promiscuity filled intercellular space; In group DMYW, collagen fiber formation, fine and messy, scattered.2The contents of IL-1α, treatment3d, Yiqi group was significantly higher than the blank and model group(P<0.05), Yiqi group also higher than whole formula and Huoxue group(P<0.05); treatment5d, whole formula group were significantly higher than the blank group and model group(P<0.05); treatment7d, Each treatment group was significantly higher than the blank group and model group(P<0.05),the whole formula group was higher than the Yiqi and Huoxue group; treatment14d, Model group and Huoxue group less than the blank group(P<0.05). The contents of TXB2, treatment3d, Normal group and the whole formula group higher than the blank group(P<0.05); treatment5d, whole formula group was higher than the blank group and the normal group(P<0.05); treatment7d, Yiqi group was higher than the blank and model group(P<0.05),also higher than the whole formula and Huoxue group(P<0.05);treatment14d, Huoxue was higher than the blank and model group(P<0.05),the blank group and normal group was less than these treatment groups(P<0.05).3At the different phase of granulation tissue gene expression, Dosing3d, each group compared with the model group were statistically significant (P<0.05), there are significant differences (P<0.05) Qi group and the control group, the normal group, the whole side group than, blood group and the control group, the whole side group than there are significant differences (P<0.05); administration7d, model group and the groups were statistically significant compared (P<0.05), a wholly-party group and the control group, and Qi group than in the normal group there were significant differences (P<0.05), promoting blood group and the control group, the normal group, Qi group and model group were significantly different (P<0.05); administration14d, model group and all-party group, Qi group, blood group than there are significant differences (P<0.05), all-party group and the control group, there was significant difference than the normal group (P<0.05).Expression of eNOS gene content, The administration of3D, the group eNOS was higher than that in the model group, the difference has statistical significance (P<0.05);7D medication, dosing groups eNOS content above the model group, a statistically significant difference(P<0.05); The normal group was higher than that of model group and Huoxue group, the difference has statistical significance (P<0.05), YiQi group than those in the normal group, the whole group and Huoxue group, the difference has statistical significance(P<0.05); The administration of14D, the Huoxue group eNOS was higher than that in the model group, the difference has statistical significance(P<0.05).Conclusion1The " STZ injection-hydrocortisone interference-skin excision-foreign body embedded " composite factors molding method can make the symptom of Yang Deficiency Rats, can prolong the time of wound healing. The wound form was similar with chronic skin ulcer that also called as yin-syndrome skin ulcer.2Huiyang Shengji ointment can improve the content of IL-1a and TXB2in serum of rats properly, Effect of Yiqi Wenyang medicine promotes inflammatory reaction in rats with immunosuppression, Huoxue Tongluo group pro-inflammatory role in anaphase.3Huiyang Shengji ointment can increase the granulation tissue eNOS gene expression, Yiqi Wenyang medicine administration early on promoting healing factor promoting expression obviously, Huoxue Tongluo group in the administration of post healing effect.