OBJECTIVE: To investigate the effectiveness and preliminary mechanisms of icariin (ICA) in enhancing the reparative effects of adipose-derived stem cells (ADSCs) on skin radiation damagies in rats. METHODS: Twelve SPF-grade Sprague Dawley rats [body weight (220±10) g] were subjected to a single dose of 10 Gy X-ray irradiation on a 1.5 cm×1.5 cm area of their dorsal skin, with a dose rate of 200 cGy/min to make skin radiation damage model. After successful modelling, the rats were randomly divided into 4 groups ( n=3), and on day 2, the corresponding cells were injected subcutaneously into the irradiated wounds: group A received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL), group B received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL)+1 μmol/L ICA (0.1 mL), group C received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a hypoxia-inducible factor 2α (HIF-2α) inhibitor+1 μmol/L ICA (0.1 mL), and group D received 0.1 mL of rat ADSCs (1×10 ⁷cells/mL) pretreated with a Notch1 inhibitor+1 μmol/L ICA (0.1 mL). All treatments were administered as single doses. The skin injury in the irradiated areas of the rats was observed continuously from day 1 to day 7 after modelling. On day 28, the rats were sacrificed, and skin tissues from the irradiated areas were harvested for histological examination (HE staining and Masson staining) to assess the repair status and for quantitative collagen content detection. Immunohistochemical staining was performed to detect CD31 expression, while Western blot and real-time fluorescence quantitative PCR (qRT-PCR) were used to measure the protein and mRNA relative expression levels of vascular endothelial growth factor (VEGF), platelet-derived growth factor BB (PDGF-BB), fibroblast growth factor 2 (FGF-2), interleukin 10 (IL-10), transforming growth factor β (TGF-β), HIF-2α, and Notch1, 2, and 3. RESULTS: All groups exhibited skin ulcers and redness after irradiation. On day 3, exudation of tissue fluid was observed in all groups. On day 7, group B showed significantly smaller skin injury areas compared to the other 3 groups. On day 28, histological examination revealed that the epidermis was thickened and the dermal fibers were slightly disordered with occasional inflammatory cell aggregation in group A. In group B, the epidermis appeared more normal, the dermal fibers were more orderly, and there was an increase in new blood vessels without significant inflammatory cell aggregation. In contrast, groups C and D showed significantly increased epidermal thickness, disordered and disrupted dermal fibers. Group B had higher collagen fiber content than the other 3 groups, and group D had lower content than group A, with significant differences ( P<0.05). Immunohistochemical staining showed that group B had significantly higher CD31 expression than the other 3 groups, while groups C and D had lower expression than group A, with significant differences ( P<0.05). Western blot and qRT-PCR results indicated that group B had significantly higher relative expression levels of VEGF, PDGF-BB, FGF-2, IL-10, TGF-β, HIF-2α, and Notch1, 2, and 3 proteins and mRNAs compared to the other 3 groups ( P<0.05). CONCLUSION ICA may enhance the reparative effects of ADSCs on rat skin radiation damage by promoting angiogenesis and reducing inflammatory responses through the HIF-2α-VEGF-Notch signaling pathway.
Animals ; Rats, Sprague-Dawley ; Skin/pathology* ; Rats ; Vascular Endothelial Growth Factor A/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Signal Transduction ; Flavonoids/pharmacology* ; Adipose Tissue/cytology* ; Stem Cells/cytology* ; Receptors, Notch/metabolism* ; Radiation Injuries, Experimental/metabolism* ; Wound Healing/drug effects* ; Male

OBJECTIVETo study the effects of Zhuhong ointment on accumulation in the body of mercury and the pathological morphology changes of kidney, via the measurement of related indicators of the skin-impaired model rat.

METHODEighty-eight SD rats were randomly divided into the impairment control group, and high-, middle-, low-dose Zhuhong ointment groups. Each group was treated by corresponding methods for 4 weeks, and recovering for 4 weeks. Urinary potein (PRO), pH, Beta N-acetyl aminoglycosidase enzymes (NAG) and beta2-microglobulin (beta2-MG) contents in urine were taken as monitoring indexes, blood urea nitrogen (BUN) and serum creatinine (SCr) in blood and the levels of mercury in urine, blood and kidney were tested, and the pathological morphology changes of kidney were observed.

RESULTAfter treatment for 4 weeks, compared with impairment control group, the levels of mercury in urine, blood and kidney in every dose group increased significantly (P < 0.01). And the relation exists between toxicity and dose on Zhuhong ointment. After recovery for 4 weeks, the levels of mercury in urine and blood in every dose group restore normal, while the level of mercury in kidney in high- dose group still increased (P < 0.01). The level of NAG increased only in high-dose group. There was no significant difference in NAG contents between Zhuhong ointment groups and the impairment control group (P < 0.05).

CONCLUSIONExcess using Zhuhong ointment repeatedly may lead to accumulation of mercury and pathological morphology changes of kidney. So the levels of mercury in the body and related indicators of renal functions should be tested in clinical when long-term using Zhuhong ointment.

Acetylglucosaminidase ; drug effects ; urine ; Animals ; Blood Urea Nitrogen ; Creatinine ; blood ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; toxicity ; Female ; Hydrogen-Ion Concentration ; drug effects ; Kidney ; drug effects ; enzymology ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; urine ; Ointments ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Retinol-Binding Proteins ; drug effects ; urine ; Skin ; drug effects ; injuries ; Time Factors ; beta 2-Microglobulin ; urine

OBJECTIVETo observe the effect of single administration of mercury- containing preparation Jiuyi Dan (calcined gypsum-Shengdan 9: 1) and Shengdan on acute toxicity of rabbits, in order to assess the safety of tested drugs.

METHODThe rabbits were randomly divided into 4 groups: the calcined gypsum group (excipient control), the Jiuyi Dan group, the 90 mg Shengdan group and the 180 mg Shengdan group. After 270 mg of calcined gypsum, 300 mg of Jiuyi Dan, 90 mg of Shengdan, and 180 mg of Shengdan were used on the surface of wounds (5 cm x 5 cm) on two sides of rabbit back for 5 h, the surfaces of wound were washed by water. The bloods were taken from the rabbit hearts before and after the drug administration for 24 h, 72 h, 7 d and 14 d for determining Hg level in blood and liver & kidney function indicators (ALT, AST, CREAT, and BUN). The rabbits were dissected after the drugs treatment for 14 d, and pathological tests were made for their livers and kidneys.

RESULTCompared with the calcined gypsum group, the 90 mg Shengdan group and the 180 mg Shengdan group showed significant increase (P < 0.01 or P < 0.05), as evidenced by increase in CREAT for 24 h and 72 h and increase in BUN for 24 h and on 7 d. AST is significantly increased as well (P < 0.01) for 24 h and 72 h compared to that of the group before drug treatment. The Hg level in blood was significantly enhanced (P < 0.01) after the rabbits were administrated with drugs for 24 h to 72 h. The pathological changes in livers and kidneys of rabbits were observed in the two doses of Shengdan treatment groups.

CONCLUSIONThe Hg blood levels were increased significantly in an obvious dose-effect relationship in all drugs treatment groups. Liver & kidney function indicators were influenced by Shengdan treatment to some extent. Meanwhile, pathological changes in rabbit livers and kidneys were also caused by Shengdan, while Jiuyi Dan has no significantly effect on livers and kidneys.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Blood Urea Nitrogen ; Body Weight ; drug effects ; Creatinine ; blood ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Female ; Kidney ; drug effects ; metabolism ; pathology ; Liver ; drug effects ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; urine ; Rabbits ; Random Allocation ; Skin ; drug effects ; injuries ; Time Factors ; Toxicity Tests, Acute

OBJECTIVETo study the effect of Badu Shengji San (BDSJS) and its decomposed recipes on morphological changes of injured skin in rats.

METHODSD rats with injured skin were treated with BDSJS and its different decomposed recipes for consecutively 14 days. Morphological changes in the injured skin were observed by H&E staining.

RESULTMercury and lead-containing ingredients significantly decreased epidermal thickness and caused vascular hemorrhage, hyperemia and inflammatory cell infiltration in reticular layer of dermis. The compatible herbs alleviated epidermal thickness and reduced dermal lesions.

CONCLUSIONBDSJS' mercury and lead-containing ingredients can accelerate the healing of skin wound and its compatible herbs can relieve the dermis injury induced by mercury and lead.

Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; toxicity ; Epidermis ; drug effects ; injuries ; pathology ; Hemorrhage ; chemically induced ; Hyperemia ; chemically induced ; Lead ; toxicity ; Male ; Mercury ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; injuries ; pathology ; Skin Diseases ; drug therapy ; pathology ; Wound Healing ; drug effects

OBJECTIVETo compare the effects of Badu Shengji San (BDSJS) on rats with different injured skins.

METHODThe injured and ulcerous skin rat model was established to observe the renal injury induced by BDSJS, a mercury-containing external preparation of Chinese medicine, with urinary N-acetyl-beta-D-glucosaminidase (NAG) and retinol binding protein (RBP) as indicators of renal toxicity.

RESULTCompared to injured skin rats with the same dose, both of high and low-dose ulcerous skin groups showed obvious increase in urinary RBP and kidney coefficients, significant pathomorphological changes in renal tubules and notable epithelial cytopathic effects. In terms of NAG, the high-dose ulcerous skin group saw no significant increase, but the low-dose group recorded sharp rise.

CONCLUSIONThe renal toxicity induced by BDSJS in ulcerous skin rats was more toxic than that in injured skin ones.

Acetylglucosaminidase ; urine ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Mercury ; toxicity ; urine ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Retinol-Binding Proteins ; urine ; Skin ; drug effects ; injuries ; Skin Ulcer ; drug therapy ; microbiology ; Staphylococcal Skin Infections ; drug therapy

OBJECTIVETo compare the sensitivity of early renal injury induced by mercury-containing medicine in rats, including urinary N-acetyl-beta-D-glucosdminidase (NAG), beta2-microglobulin (beta2-MG), retinol binding protein (RBP) and clusterin (CLU).

METHODBadu Shengji San(BDSJS), a mercury-containing preparation of traditional Chinese medicine, was adopted as the mercury contact drug. The lowest effective toxic dose was used to observe its effect on serum creatinine (SCr), blood urea nitrogen (BUN), and such early renal injury indicators as NAG, RBP, beta2-MG and CLU and compare the sensitivity of tested indicators.

RESULTCompared to the broken skin group, groups with administration of 60 and 120 mg x kg(-1) doses of BDSJS showed no obvious difference in SCr and BUN when kidney indicators is remarkably increased and obvious pathological changes were found in kidney tubules but with significant increase in the urinary level of CLU and the levels of NAG and RBP. H&E staining of renal tubule showed that exposure of 30 mg x kg(-1) BDSJS had no significant morphological changes, but at the same concentrations, the level of RBP was markedly increased. Urinary beta2-MG levels were markedly decreased in BDSJS 30, 60 mg x kg(-1) group rats, whereas 120 mg x kg(-1) dose group showed no obvious change in urinary beta2-MG levels.

CONCLUSIONUrinary RBP, NAG and CLU were more sensitive than SCr and BUN as indicators for early renal injury in the order of RBP > NAG > CLU, and urinary RBP, NAG would increase earlier than beta2-MG.

Acetylglucosaminidase ; urine ; Animals ; Blood Urea Nitrogen ; Clusterin ; urine ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; toxicity ; urine ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Retinol-Binding Proteins ; urine ; Skin ; drug effects ; injuries ; Time Factors ; beta 2-Microglobulin ; urine

OBJECTIVETo study the mercury accumulation in injured skin rats induced by Badu Shengji San (BDSJS), a traditional Chinese medicine preparation for external use.

METHODInjured skin rats were treated with BDSJS for consecutively 4 weeks. During the 4 weeks and the following 4 weeks after the drug withdrawal, samples were collected for determining mercury contents in blood, urine and kidney, with urinary N-acetyl-beta-D-glucosaminidase(NAG) and beta2-microglobulin (beta2-MG) as indicators of renal toxicity and serum biochemical indicators of hepatic and renal functions. Additionally, activated partial thromboplastin time (APTT), prothrombin time (PT) and kidney and renal pathological changes were also observed.

RESULTCompared to injured skin rats, mercury contents of blood, urine and kidney were increased significantly in low, middle and high-dose BDSJS groups administered for consecutive 4 weeks. The levels of mercury showed decreases in urine (89%, 78%, 93%) and kidney (55%, 51%, 57%), and blood mercury concentration recovered to the normal range in low, middle and high-dose BDSJS groups after the drug withdrawal for 4 weeks. Kidney coefficient and beta2-MG were remarkably increased and renal tubular epithelial cell swelling could be found in the high-dose group, and kidney coefficient, beta2-MG and renal morphology basically recovered to the normal levels after the drug withdrawal for 4 weeks.

CONCLUSIONThe administration of BDSJS for consecutively 4 weeks can cause mercury accumulation in blood and mainly in kidney. Once the accumulated mercury concentration of kidney reaches a certain level, renal tubular epithelial cells would be injured. 1.1 mg x cm(-2) of BDSJS is proved to be safe and 2.2 mg x cm(-2) can cause mild but reversible injury in the function of kidney which can be recovered after drug withdrawal for 4 weeks.

Acetylglucosaminidase ; urine ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Kidney Tubules ; drug effects ; metabolism ; pathology ; Male ; Mercury ; blood ; metabolism ; toxicity ; urine ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; injuries ; Time Factors ; beta 2-Microglobulin ; urine

OBJECTIVETo evaluate the effects of topical DMSO and intralesional hyaluronidase administration, used alone or in combination, on skin injury due to vinorelbine extravasation in rats.

METHODSSkin injury due to vinorelbine extravasation was induced in the lower extremities of 30 SD rats, which were treated subsequently with topical DMSO, intralesional hyaluronidase, their combination, topical saline, and intralesional saline, with the rats without any treatment as the control. The wound area on 1, 4, 8, 12, 18, 24, 30 days and the time of healing were observed and compared.

RESULTSThe wound area on 1, 4, 8, 12, 18, and 24 days were significantly smaller in topical DMSO group than in topical saline and control groups (P<0.05), and so in intralesional hyaluronidase group than in intralesional saline and control groups (P<0.05), but there was no significant difference between single agent (hyaluronidase and DMSO) treatment group and the combined treatment group. The healing time was significantly shorter in topical DMSO and intralesional hyaluronidase groups than in topical and intralesional saline groups and control group ( 24.9-/+3.2 and 21.9-/+3.0 days vs 29.8-/+2.6, 28.6-/+4.1 and 30.6-/+3.0 days, P<0.01), but comparable between the two single agent groups and combined treatment group (23.3-/+3.8 days).

CONCLUSIONIntralesional hyaluronidase and topical DMSO application are effective therapies for skin damage due to vinorelbine extravasation, and their combination does not improve the therapeutic effect.

Administration, Topical ; Animals ; Dimethyl Sulfoxide ; administration & dosage ; pharmacology ; Drug Therapy, Combination ; Female ; Hyaluronoglucosaminidase ; administration & dosage ; pharmacology ; Injections, Intralesional ; Male ; Rats ; Skin ; drug effects ; injuries ; pathology ; Time Factors ; Vinblastine ; analogs & derivatives ; pharmacology ; Wound Healing ; drug effects

OBJECTIVETo explore the molecular biological mechanism of arnebia root oil in promoting wound surface healing by observing histological change and basic fibroblast growth factor(bFGF) mRNA expression in the wound surface tissues of 2 groups, as well as the wound surface healing rate.

METHODExperimental model of incised-wound was produced on the back of 18 New Zealand albino rabbits. The wound surfaces were randomly divided into two groups, namely, experimental group and control group. The wound surfaces in the experimental group were treated by arnebia root oil and those in control group were treated by petrolatum gauze. Then raw surfaces were evaluated by the techniques of histology, histochemistry and electron microscope and the healing rates of the raw surfaces were compared between the two groups. Content of bFGF and it's mRNA expression in wound surface tissue was also measured by means of Western-blot and RT-PCR.

RESULTThe wound surface healing rate in experimental group was higher than that in control group( P < 0.05). The fibroblast, collagen and blood capillaries were comparatively richer in experimental group as compared with those in control group, and similarly, the expression of bFGF mRNA was also significantly enhanced in the experimental group as compared with control group during the various periods of treatment. In addition, the changes in the expressions of bFGF and it's mRNA paralleled the changes of healing rates in the two groups.

CONCLUSIONthe present results showed that amebia root oil significantly can promote the healing of raw surfaces, which may be mediated by up-regulation of bFGF expression.

Animals ; Boraginaceae ; chemistry ; Female ; Fibroblast Growth Factor 2 ; biosynthesis ; genetics ; Male ; Plant Oils ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Random Allocation ; Skin ; injuries ; metabolism ; pathology ; Wound Healing ; drug effects ; Wounds and Injuries ; metabolism ; pathology

Dermal wound healing is a complex process that involved inflammation leading to re-epithelialization, granulation tissue, and tissue remodeling. Previous studies from our laboratory have shown that polysaccharides isolated from fungus, Phellinus gilvus (PG) have various anti-inflammatory activities. In present study, we have assessed the effect of polysaccharides from PG on the dermal wound healing of polysaccharides from PG in streptozotocin-induced diabetic rat model. Six of 6-mm circular wounds were created with biopsy punch on the 4th day after induction of diabetes. After 24 hours, each test substance was applied to the wound twice a day for next 5 days. Circular wounds treated with PG showed significantly reduced wound contraction and complete reepithelialization, as compared to wounds of non-treated (p < 0.05). These results show that polysaccharides isolated from PG enhanced wound repair in diabetic impaired healing, and could be developed as a wound healing agent in such clinical settings.
Administration, Cutaneous ; Animals ; Anti-Inflammatory Agents/pharmacology ; Basidiomycota/*metabolism ; Diabetes Mellitus, Experimental/*pathology ; Histocytochemistry ; Male ; Polysaccharides/isolation&purification/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Skin/*injuries ; Streptozocin ; Wound Healing/*drug effects ; Wounds, Penetrating/*drug therapy