Effects of angiotensin II type 2 receptor blocker on wound healing in a mouse skin full-thickness injury model

AIM：To observe the effect of angiotensin II (Ang II) and its type 2 receptor (AT2R) on re-epithelialization, granulation tissue formation and growth factor production during wound healing, and to explore the possible mechanism by which Ang II and AT2R influence wound healing. METHODS：Two full-thickness skin wounds were created on the dorsum of C57BL/6J mice. The animals were treated with or without AT2R blocker PD123319 at a dose of 10 mg/kg daily after wounding. Specimens were taken from the wound of each mouse on days 3, 5, 7, 9, 11, 13 and 15 after wounding. Re-epithelialization and granulation tissue formation in the wounded skin tissues were evaluated by HE staining. The production of growth factors,epidermal growth factor (EGF),vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in wounded tissues during the healing process was detected by ELISA. RESULTS：Treatment with PD123319 significantly increased the rate of re-epithelialization and the area of granulation tissue compared with control group at 5 d and 7 d after wounding. Moreover, peritoneal application of PD123319 increased the production of EGF, VEGF and bFGF in the wounded tissues at the indicated time points after wounding. CONCLUSION：AT2R blocker PD123319 accelerates wound healing via promoting re-epithelialization,granulation tissue formation and growth factor production.     

Effects of bFGF on neuronal apoptosis and fractalkine expression in cerebral ischemia/reperfusion rats

AIM: To study the effects of basic fibroblast growth factor (bFGF) on neuronal apoptosis and fractalkine expression in ischemic penumbra after cerebral ischemia/reperfusion in rats.METHODS: Thirty-six rats were randomly divided into 3 groups: sham operation group, ischemia/reperfusion group and bFGF group. The model of middle cerebral artery occlusion was established by the method of intraluminal filament blockage. The middle cerebral arteries were blocked for 1 h and then reperfused for 24 h. Neurological performances of all rats were scored with Bederson's standard. The brain tissues of the rats were stained and the average infarct volume was calculated. TUNEL method was used to determine the number of apoptotic neurons, and the expression of fractalkine was detected by the method of immunohistochemistry.RESULTS: The score of neurological performances in bFGF group was 2.23±0.59, lower than that in ischemia/reperfusion group (3.18±0.65). The number of apoptotic neurons in bFGF group (13.22±1.35) was lower than that in ischemia/reperfusion group (17.28±1.01, P<0.05), which was the lowest in sham operation group (0.91±0.65). Compared with sham operation group, the expression of fractalkine in ischemia/reperfusion group was decreased. The expression of fractalkine in bFGF group was mainly higher than that in ischemia/reperfusion group (P<0.05).CONCLUSION: Up-regulation of fractalkine may be one of the molecular mechanisms of bFGF to protect neurons against ischemia/reperfusion injury.     

Coriaria sinica Maxim extract promotes burn wound healing in rats

AIM：To investigate the effects of Coriaria sinica Maxim extract (CSME) on burn wound healing in rats. METHODS：Forty male SD rats were randomly divided into normal saline (NS) group, white petrolatum jelly (WPJ) group, silver sulfadiazine (SSD) group and CSME group. After the animals were anesthetized, the skin of their backs was burnt to induce deep Ⅱ degree burn wounds. These wounds were treated respectively for 21 d by covering dressings with NS, WPJ, SSD and CSME, respectively. On the 1st, 3rd, 7th, 14th and 21st days, after the clinical symptoms of the animals and conditions of the wounds such as the epithelization rate, crusting and hair growth were observed, the wound tissues were also taken for histological examination. The content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD), and the expression of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and collagen were detected. RESULTS：On the 21st day after burn, the new epithelial tissues in CSME group extraordinarily　developed and a great deal of hair also grew along the margin of the wounds. The epithelization rate of the wound tissues in CSME group was higher than that in other groups. There was rare new hair in the center of the wound area in SSD group, but intensive new hair was observed in CSME group. On the 14th day and 21st day after burn, multilayer of epithelial cells was entirely covered with the wound area in CSME group. Some of the healing signs, such as sufficient differentiation and a line up in order of collagen fibers, clear tissue structure, exceedingly active hyperplasia of sebaceous glands and hair follicles, and so on, were also observed in the wound area in CSME group. From the 1st day to 21st day after burn, the increase in protein expression of EGF and bFGF in the wound tissues was significantly higher than that in other groups in the early stage, which was quickly decreased and was obviously lower than other groups in the latter stage. The mRNA expression ratio of type I and III collagens in SSD group was significantly higher than that in other groups, while that in CSME group was significantly lower than that in other groups. CONCLUSION：CSME promotes burn wound healing without scarring. The effects of CSME are likely to associate with the expression reinforcement of EGF and bFGF at mRNA and protein levels in the early stage, and associate with the expression inhibition of them later. Moreover, CSME may also inhibit the mRNA expression of type I collagen and promote the synthesis of type III collagen in the wound tissues of burn.     

Effects of hydrogen sulfide on impaired wound healing in ob/ob mice

AIM: To investigate the role of hydrogen sulfide(H₂S) on impaired wound healing in ob/ob mice and the underlying mechanism.METHODS: The ob/ob mice were randomly divided into 3 groups, including vehicle, insulin and NaHS for treatment. C57BL/6 mice were treated with vehicle as control. Full-thickness punch biopsy wounds were created on the mice. Firstly, H₂S concentrations in the skins and granulation tissues were measured. The mRNA expression of cystathionine γ-lyase(CSE) was detected by RT-qPCR. The protein expression of CSE and MMP-9 were determined by Western blot. The neutrophil and monocyte/macrophage infiltration was analyzed by immunohistochemistry me-thod. The levels of tumor necrosis factor(TNF)-α and interleukin(IL)-6 were measured by ELISA.Collagen formation was measured by Masson staining.RESULTS: The H₂S levels in the skin and granulation were significantly decreased in ob/ob mice and increased in the NaHS-treated mice(P<0.05). CSE expression at mRNA and protein levels was significantly decreased in ob/ob mice compared with the control mice(P<0.05). The wound healing period was significantly shorter in NaHS group than that in vehicle-treated ob/ob mice group(P<0.05), in which the insulin group had no difference with vehicle ob/ob mice group. The neutrophil and monocyte/macrophage infiltration, and TNF-α and IL-6 levels were significantly increased in ob/ob groups, but were decreased in NaHS group(P<0.01 or P<0.05). Meanwhile, NaHS increased collagen formation in the granulation tissues of ob/ob mice.CONCLUSION: H₂S/CSE down-regulation contributes to impaired wound healing in diabetes, which is alleviated by exogenous H₂S possibly through anti-inflammation.     

Expression of VEGF and bFGF in rat model of pressure ulcer

AIM:To study the effects of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) on the molecular pathogenesis of pressure ulcer.METHODS:SD rats were randomly divided into control group and experiment group. The pressure ulcer model was established by magnetic disk circulating compression method. HE staining was used to observe the pathological changes of the skin in the rats. The expression of VEGF and bFGF in the tissues was detected by immunohistochemical method. RESULTS:The expression of VEGF and bFGF in the tissues of rat Ⅲ-degree pressure ulcer was lower than that in the surrounding tissues and normal skin(P<0.01). The changes of VEGF and bFGF were consistent(κ=0.58). CONCLUSION:The expression levels of VEGF and bFGF are decreased in the tissues of rat pressure ulcer, suggesting that they may be the potential key factors in the difficult healing of pressure ulcer.     

Effects of hydrogen sulfide on wound healing of skin ulcer in diabetic rats

AIM: To study the effect of hydrogen sulfide on wound healing of skin ulcer in diabetic rats.METHODS: Male SD rats were randomly divided into 3 groups, including non-diabetic control(NDC) group, untreated diabetic control(UDC) group, and treated diabetic administration(TDA) group. Diabetic rats were induced by intraperitoneal injection of streptozotocin(STZ). After 1 week, wound healing model was prepared by making a round incision(2.0 cm in diameter) on the dorsal skin in full thickness. The rats from TDA group received 2% sodium bisulfide ointment on the skin ulcer wound, and the animals from UDC and NDC groups received control cream. After 21 d of treatment with sodium bisulfide, blood samples were collected for biochemical analysis, including prothrombin time(PT), thrombin time(TT), and fibrinogen(FIB) in plasma, as well as the activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) in the serum. White blood cells(WBC) and lymphocytes were also counted. Granulation tissues from the wound were processed for histological examination and Western blot analysis was used to detect heme oxygenase-1(HO-1) and tumor necrosis factor α(TNF-α) expression.RESULTS: Compared with UDC group, sodium bisulfide treatment accelerated wound healing of skin ulcer(P<0.01), and increased the activity of SOD in serum(P<0.01) in the diabetic rats. The declined number of WBC and lymphocytes, prolonged PT and TT, and decreased FIB levels in rats treated with sodium bisulfied were also confirmed. Pathological section showed that there were inflammatory cell infiltration, and irregular and loose fibril alignment in the granulation tissue of rats from the UDC group, but there were regular fibril alignment and increased angiogenesis in the granulation tissue of rats from the TDA group(P<0.05). Furthermore, sodium bisulfide treatment raised HO-1 protein expression, and decreased TNF-α protein expression in the diabetic rats.CONCLUSION: Hydrogen sulfide accelerates the wound healing of skin ulcer in the rats with diabetes. The beneficial effect of H₂S may be associated with formation of granulation, anti-inflammation, and antioxidation.     

Effects of basic fibroblast growth factor on proliferation and collagen production in human umbilical cord mesenchymal stem cells

AIM：To observe the growth pattern and surface markers of human umbilical cord mesenchymal stem cells(hUCMSCs) and to explore the influence of basic fibroblast growth factor (bFGF) on the proliferation and collagen production in hUCMSCs cultured in vitro. METHODS：hUCMSCs were isolated by enzyme digestion method and adherent culture. The surface markers CD45, CD34, CD105, CD29 and HLA-DR of the cells were analyzed by flow cytometry. The osteogenic ability and adipogenic differentiation were confirmed with oil red O and alizarin red staining. The optimal concentration of bFGF to promote the proliferation of hUCMSCs was 20 μg/L. The cells in control group were cultured in the growth medium consisting of DMEM/F12 and 10% volume fraction of fetal bovine serum. The cells in experiment group were cultured under the same condition of control group but plus 20 μg/L bFGF. The proliferation of hUCMSCs was analyzed by MTT assay. The expression of type I and III collagens at mRNA and protein levels was determined by RT-PCR and Western blotting. RESULTS：The growth curves indicated that bFGF promoted the proliferation of hUCMSCs. The hUCMSCs expressed CD29, but did not express CD34, CD45 or HLA-DR in the presence or absence of bFGF unanimously. The cells were alizarin red staining-positive and oil red O staining-positive. Compared with control group, the expression of type I and III collagens significantly decreased at mRNA and protein levels in experiment group. CONCLUSION：bFGF promotes the proliferation of hUCMSCs and does not change the expression of the surface markers. bFGF inhibits the expression of type I and III collagens at mRNA and protein levels, indicating that bFGF enhances the healing of wound without inducing scar hyperplasia.     

Expression changes of Wnt/β-catenin signaling pathway in diabetic ulcer

AIM: To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer. METHODS: Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin following high-fat diet feeding. A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding. The expression of β-catenin, GSK-3β and Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA. RESULTS: In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group. The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3β was exactly the opposite (P<0.05). CONCLUSION: The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.     

Effect of acupuncture on expression of EGF and bFGF in brain tissues of rats with traumatic brain injury

AIM: To observe the effect of acupuncture on the expression of epidermal growth factor (EGF) and basic fibrolblast growth factor (bFGF) in the brain tissues of rats with traumatic brain injury. METHODS: Thirty SD rats were randomized into sham-operated group, model group and acupuncture group. The model of traumatic brain injury was established by free drop impact. Acupuncture was performed to the rats in acupuncture group once every day and 7 days altogether. Brain histotomy was conducted after the treatment. Immunohistochemical method was adopted to test the protein expression of EGF and bFGF. RESULTS: Compared to sham-operated group, the expression of EGF in the brain tissues of model group decreased (P<0.01), and the expression of bFGF increased (P<0.01). Compared to model group, the expression of EGF and bFGF in acupuncture group increased obviously (P<0.01). CONCLUSION: Acupuncture significantly increases the expression of EGF and bFGF, and improves the repair of injured brain tissues. This might be one of the mechanisms by which acupuncture can treat traumatic brain injury and improve the nervous function.     

Effect of bFGF on expression of endothelial cell integrin β1 subunits

AIM: To study the effect of basic fibroblast growth factor(bFGF) to the expression of integrin β₁ subunit of endothelial cells. METHOD: The expression of integrin β₁ subunit of endothelial cells was determined before and after bFGF treatment with Western Blot and immage analysis method.RESULTS: The mean gray value of immunostain by immage analysis method is 166.11±9.86 in the experimental group and 175.32±5.12 in the control group, suggested that bFGF may upregulate expression of integrin β₁ subunits of endothelial cell. CONCLUSION: bFGF may play an important role in angiogenesis by way of inducing the overexpression of endothelial cell integrin β₁ subunits.     

Effects of alginate dressing and mouse epidermal growth factor combination therapy on epidermal stem cells in patients with refractory wound

AIM: To evaluate the effect of combination therapy with alginate dressing and mouse epidermal growth factor (mEGF) on proliferation and differentiation of epidermal stem cells (ESCs) in patients with refractory wound.METHODS: Eighteen cases,12 males and 6 females,aged from 18 to 61 years (mean 36.4 years),of skin defects therapy by dressing changing for one month were selected in this study.The wounds were 11 in the foot,3 in the calf,2 in the thigh and 2 in the forearm.All the patients were randomly divided into 3 groups: alginate dressing and mEGF group (n=6),mEGF group (n=6) and control group (n=6).The wound closure indexes were measured at 7th,14th,21st and 28th days.The samples were harvested at 7th and 14th days after treatment for pathologic examination.The positive staining cells of cytokeration 10 (CK10) and cytokeration 15 (CK15) were evaluated by SP immunohistochemical staining technique.RESULTS: The wound healing was promoted in group A and group B.However,the wound closure index was increased in group A (P<0.05).Pathological examination showed the epidermis was thicker in group A than that in group B and C.Treatment by the alginate dressing combined with the mEGF,angiogenesis was active and granulation was enhanced.As compared with group B and group C,SP immuohistochemical staining technique showed that the ESCs in group A were bigger in size and larger in number.There was significant difference between group A and group B (or C) in the proliferation and differentiation of ESCs.CONCLUSION: Combined therapy with alginate dressing and mEGF shows a better result than that with mEGF only in proliferation and differentiation of ESCs of patients with refractory wound.     

Effect of basic fibroblast growth factor on radiation-induced splenocytic apoptosis of mice

AIM: To investigate the effect of basic fibroblast growth factor (bFGF) on radiation-induced splenocytic apoptosis of mice. METHODS: At 14 h after whole body irradiation with 0.5 Gy and 1.0 Gy,splenocytes were cultured with and without bFGF,and splenocytic apoptosis was quantitatively analysed by flow cytometry. Cell proliferation was determined by the method of [³H]-TdR incorporation. RESULTS: bFGF(1 μg/mL and 2 μg/mL) could reduce the rate of cell apoptosis,and promote the proliferation of splenocytes. CONCLUSION: bFGF could inhibit radiation-induced splenocytic apoptosis and promote the proliferation of splenocytes and then enhance body immunity.     

Apoptosis in diabetic foot ulcers and human fibroblast cells treated with AGEs

AIM：To investigate cell apoptosis in diabetic foot ulcers and the effect of advanced glycosylation end products (AGEs) on apoptosis in human fibroblast cells. METHODS：Diabetic foot patients (n=18) and 18 age-matched non-diabetic controls were recruited. The clinical and biochemical features were compared by statistics methods. Skin biopsies were obtained from foot. Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex (SABC) staining. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) technique was used to detect apoptosis of the skin tissues. Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose (changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h) or AGEs (150 mg/L, containing 5.6 mmo/L glucose). After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3. Other cells were trypsinized, washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis. RESULTS：Diabetic patients had higher levels of fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin A1c (HbA1c), and longer wound duration. The protein level of cleaved caspase-3 was significantly higher in diabetic group, suggesting that apoptosis was increased in diabetic skin tissues. TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group (8.4%±1.5% vs 3.8%±08%), which further confirmed that cell apoptosis was increased in diabetic foot tissues. In human fibroblasts, the levels of cleaved caspase-3 in normal group, sustained high glucose group, fluctuant high glucose group and AGEs group were 080±0.13, 1.22±0.18, 1.46±0.32 and 1.83±0.25, respectively. The apoptotic rates detected by flow cytometry were 2.43%±0.19%, 2.89%±0.51%, 3.99%±0.24% and 6.83%±0.36%, respectively. Both the level of cleaved caspase-3 and the apoptotic rate in AGEs group were higher than those in normal glucose group and sustained high glucose group. CONCLUSION：Increased apoptosis in diabetic foot ulcers is one of the most important reasons for impaired wound healing. As compared to sustained high glucose and glucose fluctuations, AGEs induce greater apoptosis in human fibroblast cells.     

Expression of bFGF mRNA in pulmonary artery of rat with chronic hypoxia and its effect on tension of rat pulmonary artery in vitro

AIM: To explore the role of basic-fibroblast growth factor (bFGF) in the development of pulmonary hypertension induced by hypoxia. METHODS: 1) The pulmonary arteries of SD rats with hypoxia for one and two weeks were isolated, from which the total RNA were extracted by acid guanidinium thiocyanate-phenol-chlorform .Then the levels of mRNA were measured by RT-PCR. 2) About 3mm-long arterial rings cut from SD rat pulmonary arterial stem were suspended between stainless steelhooks in chamber with warmed (37℃) Kreb's solution. Different concentrations of bFGF were added in a cumulative fashion into the chamber where the rings were suspended. The cumulative concentration response curve was obtained. RESULTS: 1)The levels of bFGF mRNA in pulmonary artery of rats with hypoxia were increased significantly compared with those that without hypoxia (2578±384 counts·min^-1 (control) vs 5303±756 (hypoxia) for 1 week and 4054±547 (hypoxia) for 2 weeks, P all ＜0.05). 2) bFGF at concentrations ranged from 5.56×10^-10~2.78×10^-7mol/L caused dose-dependent contraction of vessel rings of rat pulmonary artery (r=0.695,P＜0.05), with EC₅₀ being 2.62×10^-7mol/L. CONCLUSION: bFGF may play an important role in the hypoxic pulmonary hypertension.     

Recent advances in anti-basic fibroblast growth factor antibody

Basic fibroblast growth factor (bFGF) has been used in wound healing, bone healing, vascular grafting, lens regeneration and limp regeneration. Anti-bFGF antibody is thought to be an major important reagent for bFGF research. This review summarizes the development of anti-bFGF antibody in recent years including preparation, screening, identification and application in order to provide reference to the studies of this field in our country.     

Effects of bFGF on brain edema,nerve function injury and autophagy related protein in rats with traumatic brain injury

AIM:To study the effects of basic fibroblast growth factor (bFGF) on brain edema, nerve function damage and autophagy related proteins in rats with head injury. METHODS:The rat model of craniocerebral injury (CI) was constructed. The rats were divided into control group, CI group, and low-, middle-and high-dose bFGF groups (n=10). The CI model was established in CI group, while the rats in control group were not given epidural impact. The rats in low-dose, middle-dose and high-dose bFGF groups were given bFGF at 2, 4 and 6 μg, respectively, by intraperitoneal injection after 30 min. The neurological function in the rats was evaluated by improved neurological function scoring. The rat brain tissues were taken, and the water content was detected. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in the brain tissue were measured by ELISA. The malondialdehyde (MDA) content was analyzed by thiobarbituric acid method. The activity of superoxide dismutase (SOD) was examined by WST-8 assay. The glutathine peroxidase (GSH-Px) activity was detected by colorimetric method. The protein levels of autophagy related proteins LC3-Ⅱ and beclin-1 in the brain tissues were determined by Western blot. RESULTS:The neurological function score was increased significantly of the rats in CI group. The rat model of craniocerebral injury was successfully constructed. Neurological function scores in the rats in low-dose, middle-dose and high-dose bFGF groups were reduced, the water content of the brain tissue was also reduced (P<0.05). The levels of TNF-α, IL-6 and IL-1 β were decreased in the brain tissues (P<0.05), the content of MDA was declined (P<0.05), the activities of SOD and GSH-Px were increased (P<0.05), the protein levels of LC3-Ⅱ and beclin-1 were decreased, compared with the untreated rats in CI group (P<0.05). CONCLUSION:bFGF improves the nerve function of the rats with craniocerebral injury, reduces the water content of the brain tissue, reduces the expression of autophagic protein LC3-Ⅱ and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.     

Decreased fibroblast proliferation and collagen content in the wound of diabetic rats caused by streptozotocin

AIM: To study fibroblast proliferation and collagen synthesis during wound healing in diabetic rats induced by streptozotocin. METHODS: 30 Wistar male rats were randomly divided into control group and model group. 55 mg/kg STZ were given intraperitoneally to model rats. After 3 weeks, a round skin of 2.04 cm2 was excised on all dorsal back of rats. The healing time and healing rate were observed according to re-epithelization. The numbers of fibroblasts and the expression of proliferating cell nuclear antigen (PCNA) were observed by Hematoxylin-Eosin (HE) staining and immuno-histochemistry assay. Collagen Ⅰ and Ⅲ stained by Picric acid-Sirius red were calculated by image analysis. RESULTS: The healing time in model group was (27.13±1.81) days，significantly longer than that in control group ［(15.25±1.67） days， P<0.01］. The healing rates in model group were significantly less than that in control group at day 3, day 7 and day 15 (P<0.01). The amount of fibroblasts and the expression of PCNA in model group were significantly less than those in control group on day 3, day 5, day 7 and day 9, respectively (P<0.05, P<0.01). Even the content of collagen I in the wound of both groups increased with time, the values were much higher than that in model group at different times (P<0.05), respectively. For model group, the ratio of collagen Ⅰ/Ⅲ was less than that in control group 3, 7 and 11 days after wound (P<0.01). CONCLUSION: STZ impaires wound healing in rats, which is possible caused by the disturbance of fibroblast proliferation and collagen synthesis in the wound.     

Adenosine promotes bFGF protein and bFGF mRNA expression in human umbilical vein endothelial cells in vitro

AIM: To investigate the influence of adenosine on human umbilical vein endothelial cells (HUVEC) bFGF protein production and bFGF mRNA expression. METHODS: Immunohistochemistry staining was performed to detect bFGF protein. RT-PCR was performed to detect bFGF mRNA expression. RESULTS: Immunohistochemistry study demonstrated that there was only a small amount of bFGF positive cells and the color was weak in control group (without adenosine). In groups treated with 10-4 mol/L and 10-6 mol/L adenosine, bFGF protein was significantly higher than that in control group (P<0.05). In 10-8 mol/L and 10-10 mol/L adenosine groups, there were no significant differences compared with control group (P>0.05). RT-PCR showed that in 10-4 mol/L and 10-6 mol/L adenosine groups, bFGF mRNA expression was higher than that in control group (P<0.05), while the difference between 10-8 mol/L adenosine group and control group was not significant (P>0.05). CONCLUSION: Adenosine may promote HUVEC proliferation and angiogenesis partly through inducing bFGF expression.     

Imbalance between matrix metalloproteinases and tissue inhibitor of metalloproteinases during wound healing in diabetic rats

AIM: To investigate the imbalance between the expression of metalloproteinases (MMPs) and that of tissue inhibitors of metalloproteinase (TIMPs) during wound healing in diabetic rats. METHODS: Diabetic rats were induced with streptozotocin. All rats were maintained for 6 weeks. A full-thickness excisional wound was created on the back of each rat. Every group was randomly divided into 3 subgroups of 7 rats: 3 d group, 7 d group, 14 d group and animals were killed at 3rd, 7th and 14th day. Routine pathological examination, Masson′s trichrome staining and immunohistochemistry were made to calculate the score of epidermal and dermal regeneration, granulation tissue thickness, angiogenesis, matrix density, and infiltrated cells at different time points. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin at those time points. RESULTS: Six weeks after streptozotocin treatment, Three days after injury, the wound healing rate of normal rats was faster than that of diabetic rats. From 3rd to 14th day, there were a lot of fibroblast and macrophage in normal skin, while few such cells were observed in diabetic skin. The other histological scores in normal skin were higher than those in diabetic rats at 7th and 14th day. Both MMP-9 and TIMP-1 had minimally detectable levels before wounding but exhibited rapid, significantly large increases within 3 d after wounding. Subsequently, they showed a rapid decline by 14 d. The relative values of expression of MMP-9 mRNA and protein in diabetic group were higher than those in normal group at different time points. However, the values of TIMP-1 mRNA and protein in diabetic group were significantly lower than those in control group. Significant difference was observed between two groups with the ratio of MMP-9/TIMP-1, higher in diabetic group than that in normal group. CONCLUSION: Abnormal reepithelialization, angiogenesis, inflammatory cell infiltration, collagen fibers generation, granulation tissue deposition, seem to be the basic histopathology that delays wound healing. The imbalance between MMPs and TIMPs in diabetic skin tissue before and after injury may be one of the important reasons of these alterations of histopathology.     

Endogenous basic fibroblst growth factor is a protective factor against myocardial ischemia/reperfusion injury of rats

AIM: To observe the role of exogenous and endogenous basic fibroblst growth factor (bFGF) on myocardial ischemia/reperfusion(I/R) injury of rats.METHODS:bFGF and bFGF antiserum were applied to rat isolated I/R heart. Myocardial function, coronary effluent volume,protein and myoglobin content as well as LDH activity in coronary effluent fluid, myocardial calcium, MDA and ATP concentration as well as PKC, MAPK activity were measured. RESULTS:Compared with control, myocardial function in I/R group significantly decreased. Protein, myoglobin content and LDH activity in coronary effluent liquid as well as myocardial MDA and calcium content increased, while myocardial ATP concentration decreased(all P＜0.01). Compared with I/R group, ±LV dp/dt_max in bFGF group increased by 43% and 26%, respectively. LVEDP decreased by 40%. HRr/HRi and B/A augmented by 42% and 20%, respectively. Protein and myoglobin content as well as LDH activity lowered by 29%,30％ (all P＜0.01) and 33％ (P＜0.05) respectively. Myocardial MDA and calcium content decreased by 44% and 35%, respectively, while myocardial ATP level as well as PKC and MAPK activity increased by 34%,41％ and 10% (all P＜0.01), respectively. In bFGF antiserum group, ±LV dp/dt_max were 35% and 38% lower than those in I/R group. LVEDP increased by 93%. HRr/HRi and B/A decreased by 36% and 45%, respectively. Protein and myoglobin content as well as LDH activity augmented by 54%,96％ (all P＜0.01) and 34％ (P＜0.05) respectively. Myocardial MDA and calcium content increased by 24% and 50%, respectively, while myocardial ATP level as well as PKC and MAPK activity lowered by 28%,21％ and 8% (all P＜0.01), respectively. CONCLUSION:Endogenous bFGF is a protective factor against myocardial ischemia/reperfusion injury of rats.