Network Pharmacology Study on Herb Pair Bletilla striata-Galla chinensis in the Treatment of Chronic Skin Ulcers.

BACKGROUND: Herb pair Bletilla striata-Galla chinensis (BS-GC) is a classic combination of topical traditional Chinese medicine formulae in the treatment of chronic skin ulcers (CSUs). OBJECTIVE: The aim of this study is to explore the effective active ingredients of BS-GC, as well as the core targets and signal transduction pathways of its action on CSUs. METHODS: The ingredients of BS-GC were obtained from TCMSP and HERB databases. The targets of all active ingredients were retrieved from the SwissTargetPrediction database. The targets of CSUs were obtained from OMIM, GeneCards, Drugbank, and DisGeNET databases. A drug-disease target protein-protein interaction (PPI) network was constructed to select the most core targets, and an herb-ingredient-target network was built by utilizing Cytoscape 3.7.2. Furthermore, we performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes database (KEGG) analysis and verified the results of network pharmacology through molecular docking. RESULTS: A total of 40 active ingredients from the herb pair BS-GC were initially screened, and a total of 528 targets were retrieved. Meanwhile, the total number of CSU targets was 1032. Then, the number of common targets between BS-GC and CSUs was 107. The 13 core targets of herb pair BS-GC with CSUs were filtered out according to the PPI network, including AKT1, TNF, EGFR, BCL2, HIF1A, MMP-9, etc. The 5 main core active ingredients were 1-(4-Hydroxybenzyl)-2-methoxy-9,10-dihydrophenanthrene-4,7-diol, 1-(4- Hydroxybenzyl)-4-methoxy-9,10-dihydrophenanthrene-2,7-diol, physcion, dihydromyricetin, and myricetin. The main biological processes were inflammation, oxidative stress, and immune response, involving the AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, NF-κB signaling pathway, and calcium signaling pathway. Molecular docking results showed good binding activity between the 5 main core active ingredients and 13 core targets. CONCLUSION: This study predicted the core targets and signal transduction pathways in the treatment of CSUs to provide a reference for further molecular mechanism research.

Melatonin loaded lecithin-chitosan nanoparticles improved the wound healing in diabetic rats.

Wound healing in diabetic patients remains a worldwide problem that can cause amputations and even lead to death. This work aimed to produce lecithin-chitosan nanoparticles loaded with melatonin (MEL-NP) incorporated in a topical formulation to be evaluated for healing in the in vivo animal model for diabetes. To produce nanoparticles, an ethanolic solution containing soybean lecithin and melatonin was added dropwise to an aqueous solution of chitosan under sonication. The nanoparticles were physicochemical characterized and evaluated in vivo for toxicity using the Galleria mellonella model and its potential for wound healing in diabetic rats. The MEL-NPs presented a particle size of 160 nm and a zeta potential of 25 mV. The melatonin entrapment efficiency was 27%. Our results indicated that treatment with MEL-NP improved wound healing demonstrated by wound closure earlier than the other treatments evaluated. A desired therapeutic effect was achieved by MEL-NP in the induction of fibroblast and angiogenic proliferation. In addition, it was accompanied by an expressive collagen deposition. Considering the observed data, the MEL-NP developed could be used as a proof of concept to develop a promising strategy for the healing of diabetic wound.

NMR-based metabolomic analysis for the effects of Huiyang Shengji extract on rat diabetic skin ulcers.

ETHNOPHARMACOLOGICAL RELEVANCE: Huiyang Shengji formula (HSF) is a compound Chinese herbal medicine prescription, and has long been used for treating chronic non-healing wounds. AIM OF THE STUDY: The purpose of this study was to provide new insight into molecular mechanisms of healing effects of the HSF treatments. MATERIALS AND METHODS: We established a rat diabetic skin ulcer (DSU) model, and assessed healing effects of four HSF treatments on DSUs by calculating wound healing rates and immunohistochemical detection of the expressions of angiogenesis-related factors in the model rats (Mod) relative to normal rats (Nor), including Huiyang extract (HE), Shengji extract (SE), Huiyang Shengji extract (HSE) and HSE associated with acupuncture (Ac-HSE). We then performed NMR-based metabolomic analyses on skin tissues of the Nor, Mod, HSE-treated, Ac-HSE-treated rats to address metabolic mechanisms underlying these effects. RESULTS: These treatments up-regulated expressions of two angiogenesis-related factors VEGF and CD31, and improved efficacy of healing DSUs, in which HSE and Ac-HSE exhibited the most significant effects. Compared with Mod, HSE and Ac-HSE groups shared four characteristic metabolites (lactate, histidine, succinate and acetate) and four significantly altered metabolic pathways with Nor. Both HSE and Ac-HSE treatments could partly reverse the metabolically disordered pathological state of DSUs to the normal state. They might improve wound healing through promoting glucose metabolism, BCAAs metabolism, and enhancing antioxidant capacity and angiogenesis in DSU tissues. Ac-HSE significantly enhanced wound healing rates compared to HSE, potentially owing to significant capacities of enhancing anti-oxidation and angiogenesis and interfering three more metabolic pathways. CONCLUSIONS: This work provides a mechanistic understanding of the healing effects of the HSE and Ac-HSE treatments on DSUs, is of benefit to improvements of the HSF treatments for clinically healing chronic non-healing wounds.

Vitamin D Ameliorates Impaired Wound Healing in Streptozotocin-Induced Diabetic Mice by Suppressing Endoplasmic Reticulum Stress.

BACKGROUND: This study is designed to investigate whether vitamin D promotes diabetic wound healing and explore the potential mechanism which may be involved in the healing process. MATERIAL AND METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with 200 µg/ml of advanced glycation end product-modified human serum albumin (AGE-HSA) and 250 mg/dl of glucose with vitamin D. Cell viability was analyzed using the CCK-8 assay, and the apoptosis rate was measured using flow cytometry. Endogenous markers of ER stress were quantified using Western blot and a real-time polymerase chain reaction. Diabetic mice were treated with vitamin D (100 ng/kg per day) for 14 days. The ulcer area and ulcerative histology were detected dynamically. RESULTS: Vitamin D administration not only decreased the apoptosis rate but also increased cell viability. Furthermore, the expression of endogenous markers of ER stress was downregulated as a result of vitamin D treatment. Vitamin D supplementation significantly accelerated wound healing of diabetic mice and improved the healing quality. Further studies showed that reduced ER stress was associated with the positive outcome. CONCLUSION: These results suggest that vitamin D may ameliorate impaired wound healing in diabetic mice by suppressing ER stress.

The Four-Herb Chinese Medicine Formula Tuo-Li-Xiao-Du-San Accelerates Cutaneous Wound Healing in Streptozotocin-Induced Diabetic Rats through Reducing Inflammation and Increasing Angiogenesis.

Impaired wound healing in diabetic patients is a serious complication that often leads to amputation or even death with limited effective treatments. Tuo-Li-Xiao-Du-San (TLXDS), a traditional Chinese medicine formula for refractory wounds, has been prescribed for nearly 400 years in China and shows good efficacy in promoting healing. In this study, we explored the effect of TLXDS on healing of diabetic wounds and investigated underlying mechanisms. Four weeks after intravenous injection of streptozotocin, two full-thickness excisional wounds were created with a 10 mm diameter sterile biopsy punch on the back of rats. The ethanol extract of TLXDS was given once daily by oral gavage. Wound area, histological change, inflammation, angiogenesis, and collagen synthesis were evaluated. TLXDS treatment significantly accelerated healing of diabetic rats and improved the healing quality. These effects were associated with reduced neutrophil infiltration and macrophage accumulation, enhanced angiogenesis, and increased collagen deposition. This study shows that TLXDS improves diabetes-impaired wound healing.

Avocado/soybean unsaponifiables: a novel regulator of cutaneous wound healing, modelling and remodelling.

We investigated the effects of avocado/soybean unsaponifiables (ASU) on the healing response of cutaneous wound defect in rats. Sixty male rats were randomly divided into three groups including control, vehicle and treatment (n = 20 in each group). A 2 × 2 cm(2) wound defect was made on the dorsum. The control, vehicle and treatment groups were treated daily with topical application of saline, cream and cream/ASU for 10 days, respectively. The wounds were monitored daily. The animals were euthanised at 10, 20 and 30 days post injury (D). The dry matter, hydroxyproline, collagen, n-acetyl glucosamine (NAGLA) and n-acetyl galactosamine (NAGAA) contents of the skin samples were measured and the histopathological and biomechanical characteristics of the samples were investigated. Statistics of P < 0·05 was considered significant. Treatment significantly increased tissue glycosaminoglycans and collagen contents at various stages of wound healing compared to controls. Treatment modulated inflammation, improved fibroplasia and produced high amounts of scar tissue at short term. At long term, treatment reduced the scar tissue size and increased the quality and rate of wound contraction and reepithelisation compared to controls. The treated lesions were more cosmetically pleasing and had significantly higher biomechanical characteristics than controls. ASU was effective in rat wound healing.

Role of nicotinic acid and nicotinamide in nicorandil-induced ulcerations: from hypothesis to demonstration.

Nicorandil, a nicotinamide ester, was first reported to be involved in the induction of oral ulcers in 1997. Since then, many reports of single or multiple nicorandil-induced ulcerations (NIUs) have been reported. We hypothesised that in the case of high-dosage nicorandil or after an increased dosage of nicorandil, nicotinic acid and nicotinamide (two main metabolites of nicorandil) cannot appropriately merge into the endogenous pool of nicotinamide adenine dinucleotide/phosphate, which leads to abnormal distribution of these metabolites in the body. In recent or maintained trauma, nicotinamide increases blood flow at the edge of the raw area, inducing epithelial proliferation, while nicotinic acid ulcerates this epithelial formation, ultimately flooding the entire scar. We demonstrate, by comparison to a control patient non-exposed to nicorandil, an abnormal amount of nicotinic acid (×38) and nicotinamide (×11) in the ulcerated area in a patient with NIUs. All practitioners, especially geriatricians, dermatologists and surgeons, must be aware of these serious and insidious side effects of nicorandil. It is critical to rapidly reassess the risk-benefit ratio of this drug for any patient, and not only for those with diverticular diseases.

[Effect of Shengji Huayu Recipe on the expression of MMP-3 and TIMP-1 in skin ulcer tissue of diabetic rats].

OBJECTIVE: To study the effect of Shengji Huayu Recipe (SHR)on the expression of MMP-3 and TIMP-1 in the skin ulcer tissue of diabetic rats. METHODS: The skin ulcer model was established in diabetic mice. Different compatibility proportions of SHR [the ratio of Shengji Recipe (SJR) to Huayu Recipe (HYR) = 2:1, 1:1, and 1:2, respectively] were used to intervene. The expression of MMP-3 protein in the skin ulcer of diabetic rats was detected by Western blot method,and TIMP-1 protein was detected by immunohistochemical assay. RESULTS: At each time point, there was no statistical difference in the blood glucose level among groups (P > 0.05). But all of them increased significantly,when compared with those of the normal wound group (P < 0.01). As for the difference between after would area treatment and before would area treatment, better effect was obtained in the SHR No. 3 group and the normal ulcer group than in the diabetic ulcer model group (P < 0.05). Results of Western blot showed that the MMP-3 protein expression was higher in the SHR No. 2 group than in the SHR No.3 group (P < 0.05). Immunohistochemical results showed that TIMP-1 protein expression was lower in the SHR No. 2 group than in the SHR No. 3 group and the diabetic ulcer model group (P < 0.05). TIMP-1 protein expression was higherin the SHR No. 3 group than in the SHR No. 2 group (P < 0.01). CONCLUSION: Using SHR No.3 was conducive to the promotion of wound healing in early wound repair stage, and using SHR No. 2 might be conducive to inhibiting the formation of pathological scar.

Modulation of inflammation by Cicaderma ointment accelerates skin wound healing.

Skin wound healing is a natural and intricate process that takes place after injury, involving different sequential phases such as hemostasis, inflammatory phase, proliferative phase, and remodeling that are associated with complex biochemical events. The interruption or failure of wound healing leads to chronic nonhealing wounds or fibrosis-associated diseases constituting a major health problem where, unfortunately, medicines are not very effective. The objective of this study was to evaluate the capacity of Cicaderma ointment (Boiron, Lyon, France) to accelerate ulcer closure without fibrosis and investigate wound healing dynamic processes. We used a necrotic ulcer model in mice induced by intradermal doxorubicin injection, and after 11 days, when the ulcer area was maximal, we applied Vaseline petroleum jelly or Cicaderma every 2 days. Topical application of Cicaderma allowed a rapid recovery of mature epidermal structure, a more compact and organized dermis and collagen bundles compared with the Vaseline group. Furthermore, the expression of numerous cytokines/molecules in the ulcer was increased 11 days after doxorubicin injection compared with healthy skin. Cicaderma rapidly reduced the level of proinflammatory cytokines, mainly tumor necrosis factor (TNF)-α and others of the TNF pathway, which can be correlated to a decrease of polymorphonuclear recruitment. It is noteworthy that the modulation of inflammation through TNF-α, macrophage inflammatory protein-1α, interleukin (IL)-12, IL-4, and macrophage-colony-stimulating factor was maintained 9 days after the first ointment application, facilitating the wound closure without affecting angiogenesis. These cytokines seem to be potential targets for therapeutic approaches in chronic wounds. Our results confirm the use of Cicaderma for accelerating skin wound healing and open new avenues for sequential treatments to improve healing.

[Effect of zhuhong ointment on renal antioxidant capability in skin ulcer model rats].

OBJECTIVE: To study the effect of repeated administration of Zhuhong ointment on renal antioxidant capability of ulcerous skin in rats, in order to further discuss the mechanism of mercury contained in Zhuhong ointment on the antioxidant capability of kidney in skin ulcer rats. METHOD: Eighty SD rats were randomly divided into eight groups: Zhuhong ointment A, B, C, D, E (1.219, 0.609, 0.305, 0.152, 0.76 g x kg(-1)) groups, the vaseline group, the ulcer model group and the impairment control group. The levels of NAG and RBP of toxicity for early kidney tubular injury and T-AOC, SOD, GSH-PX and GSH in kidney were determined after consecutive administration for 14 days. RESULT: Compared with ulcer model group, the levels of RBP in groups A, B, C and D increased, while the levels of NAG increased only in the group A. The level of T-AOC increased in groups A, B and C. The level of T-SOD increased in the group E, while it dropped down greatly in the group A. The level of GSH-PX increased in groups A, B and C. The content of GSH increased in every dose groups. CONCLUSION: Antioxidant capacity in rats can be increased in a reasonable dose of Zhuhong ointment, but some antioxidant activity can be notably inhibited by with the increase of dose.

[Effects of Chinese herbal medicine Yiqi Huayu formula on substance P expression in skin ulcers of rats with diabetes mellitus].

OBJECTIVE: To investigate the effects of Chinese herbs for replenishing qi (Yiqi) and dissolving stasis (Huayu) on substance P expression in granulation tissue of skin ulcers in rats with diabetes mellitus. METHODS: One hundred male Sprague-Dawley rats with back full-thickness skin lesion were randomly divided into five groups: ulcer control group, diabetic ulcer group, Yiqi Huayu herb group, Yiqi herb group, and Huayu herb group. Except ulcer control group, the other rats were also injected streptozotocin to induce diabetes mellitus. Rats in each group were given normal saline or herbal drugs for 15 d and the wound healing rate of the rats was observed on days 8 and 15, respectively. After treatment, the expression of substance P in granulation tissues of the rats was tested with immunohistochemical method and the immunohistochemical index was calculated. RESULTS: On the 8th and 15th days after treatment, the wound healing rate of the ulcer control group was higher than that of the diabetic ulcer group (P<0.01); the wound healing rates of all the treated groups were higher than that of the diabetic ulcer group (P<0.05); the wound healing rate of the Yiqi Huayu herb group was higher than those of the Yiqi herb group and the Huayu herb group (P<0.05). Compared with the diabetic ulcer group, the expression of substance P of the ulcer control group was increased (P<0.05); the expressions of substance P of the Yiqi Huayu herb group and the Huayu herb group were higher than that of the diabetic ulcer group (P<0.05). CONCLUSION: Yiqi Huayu herbs can promote wound healing in rats with diabetes mellitus by up-regulating the expression of substance P.

[Effect of mild-warm moxibustion on microcirculation in the raw surface tissue of chronic refractory wound in skin ulcer rats].

OBJECTIVE: To observe the effects of mild-warm moxibustion on dynamic blood flow, microvessel count (MVC)and vascular endothelial growth factor (VEGF) expression in the wound tissue of the chronic skin ulcer in rats, so as to reveal its underlying mechanism in promoting wound recovery. METHODS: A total of 104 male SD rats with skin injury were randomly divided into control group (n=8), model group (n=32), TDP (far-infrared heating device) group (n=32) and moxibustion group (n=32). Chronic refractory raw surface wound model was established by muscular injection of Hydrocortisone Sodium Succinate. For rats of the TDP and moxibustion groups, TDP irridiation and mild-warm moxibustion were applied to the raw surface, bilateral "Shenshu" (BL 23) and "Zusanli" (ST 36) for 15 min, once daily for 3, 7 and 14 days respectively. The healing rate and the healing time of raw surface of the wound were observed. The blood flow of the raw surface of the wound tissue was measured by laser Doppler flowmeter and the MVC in granulation tissue of chronic skin ulcer was counted under light microscope. VEGF expression was detected by immunohistochemistry. RESULTS: In comparison with the control group, the healing rate of the wound raw surface was significantly lower and the healing time was prolonged in the model group (P < 0.01). Compared with the model group, the healing rates on day 3, 7, 10 and 14 were significantly higher and the healing time was strikingly faster in both TDP and moxibustion groups (P < 0.01, P < 0.05), and the effects of the moxibustion group in increasing the healing rate and shortening the healing time were significantly better than those of TDP group (P < 0.01). In comparison with the model group, the blood flow volume, MVC and VEGF expression levels on day 3 and 7 were upregulated significantly in both TDP and moxibustion groups (P < 0.01, P < 0.05); while the blood flow volume, MVC and VEGF expression level in the moxibustion group and the blood flow volume and VEGF expression level in the TDP group downregulated considerably on day 14 (P < 0.01). No significant difference was found between the TDP and moxibustion groups in the MVC on day 14 after the treatment (P > 0.05). CONCLUSION: Mild-warm moxibustion can promote wound healing, which is closely with its effects in increasing blood flow and MVC, and upregulating VEGF expression in the wound granulation tissue of the chronic skin ulcer.

Effects of external application of Chinese medicine on diabetic ulcers and the expressions of ß-catenin, c-myc and K6.

OBJECTIVE: To observe the clinical efficacy of Chinese medicine (CM) treatment of Hongyou Ointment and Shengji Powder on diabetic ulcers, and to observe the influence of CM treatment on the expressions of proteins associated with the Wnt signaling pathway, such as ß-catenin, c-myc and K6. METHODS: sixty-two patients fitting the registration standards were randomly divided into the CM group (31 patients) and the Western medicine (WM) group (31 patients) by a random number table. The patients in the CM group were treated with Hongyou Ointment and Shengji Powder externally. The patients in the WM group were treated with mupirocin ointment, growth factor (bFGF), and Vaseline gauze for external use and with basic therapies. Wound-healing time and four-week healing rate were recorded. The wounds were measured by digital photography and ImageJ software. Skin biopsies were obtained from 24 patients before CM treatment and 20 patients after CM treatment. Immunohistochemical tests and semi-quantitative imaging with NIH ImageJ 1.42 software were used to analyze the changes in protein expression of ß-catenin, c-myc, and K6. RESULTS: Fifty-three patients completed the trial; four patients in the CM treatment group and five patients in the WM group dropped out. Among them, four were dissatisfied with the treatment process, two could not continue because of their jobs, two failed to complete the course of follow-up appointments, and one was diagnosed with squamous cell carcinoma during treatment. The comparison of ulcer healing rates between the two groups showed insignificant differences (P=0.77). The ulcer healing rates were 33.33% (9/27) in the CM group and 26.92% (7/26) in the WM group. However, the effective rate was significantly higher in the CM group (81.48%, 22/27) than in the WM group (57.69%, 15/26, P=0.04). The mean wound healing time was shorter in the CM group (22.71 ±5.46 days) than in the WM group (26.56 ±7.56 days, P=0.04). CM treatment was well tolerated, and there was no withdrawal due to adverse reactions. Immunohistochemical analysis in the refractory wound indicated higher expressions of ß-catenin, c-myc and K6 compared with the normal skin. ß-catenin was abnormally expressed in the nuclei of the keratinocytes and fibroblasts at the wound margins, and the expressions of c-myc and K6 were highly expressed in the full hyperplastic epidermis, especially in the granular layer (P<0.05). The expressions of these proteins decreased after CM treatment. The expression levels of ß-catenin, c-myc, and K6 proteins before and after the treatment were 101.88 ± 10.76 vs. 140.42 ±8.45; 113.27 ± 16.75 vs. 153.79 ±8.32; 90.39 ±11.07 vs. 151.29 ±7.39, respectively. CONCLUSIONS: CM treatment using Hongyou Ointment and Shengji Powder was efficient in the management of diabetic skin ulcers. The mechanism of action might be related to the inhibition of the Wnt signaling pathway.

[Effects of aloe gel on doxorubicin-induced extravasation injury in rats].

BACKGROUND AND OBJECTIVE: Aloe has preventive effects on some chemotherapy-induced extravasation injuries. This study was to investigate the effect and mechanism of aloe gel on doxorubicin-induced extravasation injury. METHODS: Sprague-Dawley (SD) rats were used to establish the extravasation injury model induced by doxorubicin. Thirty SD rats were randomly divided into three groups: control group, aloe gel group (1 g/L) and 50% magnesium sulfate group. The area of extravasation was measured and the degree of injury was observed. The injured tissues were resected from two randomly selected rats in each group on the 1st, 4th, 7th, 11th, and 18th day after treatments. Pathological morphology of the resected tissues was observed under an optical microscope after hematoxylin and eosin (HE) staining. The exosmosis skin and subcutaneous tissues of rats were resected five days after treatments. Then the wounds were interruptedly sutured. When sutures were removed on the 7th day after operation, the condition of primary wound healing and the healing time were recorded. Expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) in the exosmosis skin and subcutaneous tissues were detected by immunohistochemistry. RESULTS: The area and the degree of extravasation injury were smaller and less severe in the aloe gel and magnesium sulfate groups than in the control group (P<0.01). The rates of primary wound healing were significantly higher in the aloe gel (60.0%) and magnesium sulfate (66.7%) groups than in the control group (20.0%); while the healing time was significantly shorter in the aloe gel (9.6+/-1.64 d) and magnesium sulfate (9.33+/-1.40 d) groups than in the control group (12.13+/-2.06 d) (both P<0.01). Moreover, the expression levels of VEGF and EGFR were higher in the aloe gel group than in the control group. CONCLUSION: The preventive and therapeutic effects of aloe gel on doxorubicin-induced extravasation injury are satisfactory, which may be in relation to the up-regulation of VEGF and EGFR.

Disruption of FADS2 gene in mice impairs male reproduction and causes dermal and intestinal ulceration.

Delta-6 desaturase (D6D) catalyzes the first step in the synthesis of highly unsaturated fatty acids (HUFA) such as arachidonic (AA), docosapentaenoic (DPAn-6), and docosahexaenoic (DHA) acids, as well as the last desaturation of DPAn-6 and DHA. We created D6D-null mice (-/-), which enabled us to study HUFA deficiency without depleting their precursors. In -/-, no in vivo AA synthesis was detected after administration of [U-(13)C]linoleic acid (LA), indicating absence of D6D isozyme. Unexpectedly, all of the -/- developed ulcerative dermatitis when fed a purified diet lacking D6D products but containing ample LA. The -/- also exhibited splenomegaly and ulceration in duodenum and ileocecal junction. Male -/- lacked normal spermatozoa with a severe impairment of spermiogenesis. Tissue HUFAs in -/- declined differentially: liver AA and DHA by 95%, and a smaller decrease in brain and testes. Dietary AA completely prevented dermatitis and intestinal ulcers in -/-. DPAn-6 was absent in -/- brain under AA supplementation, indicating absence of D6D isozyme for DPAn-6 synthesis from AA. This study demonstrated a distinct advantage of the D6D-null mice (-/-) to elucidate (1) AA function without complication of LA deprivation and (2) DHA function in the nervous system without AA depletion or DPAn-6 replacement seen in traditional models.

[Effects of reinforcing qi and nourishing blood recipes on vascular endothelial growth factor and microvessel count in granulation tissue in rats with chronic skin ulcers].

OBJECTIVE: To explore the effects of reinforcing qi and nourishing blood recipes on vascular endothelial growth factor (VEGF) and the microvessel count (MVC)in granulation tissue in rats with chronic skin ulcers. METHODS: A total of 90 male SD rats with back full-thickness skin lesion were used in this study. Except for the normal control group, the rats were injected with hydrocortisone for inducing chronic skin ulcers, and were randomly divided into untreated group, Sijunzi Decoction group, Siwu Decoction group and Bazhen Decoction group. The time of wound healing was observed; immunohistochemical method and image analytical method were used to test the expressions of VEGF and MVC in granulation tissue of dermal chronic ulcers in the rats. RESULTS: In the untreated group, the time of wound healing was significantly longer than that in the normal group (P<0.01). The time of wound healing was obviously shortened in the three treated groups as compared with the untreated group (P<0.01, P<0.05). In the Sijunzi Decoction group, the time of wound healing was obviously shorter than those in Siwu Decoction group and Bazhen Decoction group (P<0.05, P<0.01). In the untreated group, the expression of VEGF and MVC were significantly lower than those in the normal control group (P<0.01). The expression of VEGF and MVC were obviously increased in the three treated groups as compared with those in the untreated group (P<0.01, P<0.05). In the Sijunzi Decoction group, the expression of VEGF and MVC were obviously lower than those in Siwu Decoction group and Bazhen Decoction group (P<0.05, P<0.01). CONCLUSION: Reinforcing qi and nourishing blood recipes can promote wound healing in rats through up-regulating the expression of VEGF in granulation tissue of dermal chronic ulcers, and inducing angiogenesis.

[Effects of resolving stagnation and promoting granulation therapy on expressions of Bax and Bcl-2 in granulation tissue of diabetic rats during wound healing].

OBJECTIVE: To study the effects of resolving stagnation and promoting granulation therapy on expressions of Bax and Bcl-2 in granulation tissue of diabetic rats during wound healing. METHODS: Seventy-two male SD diabetic rats with full-thickness skin lesion were randomly divided into 3 groups: SJHYR 1-treated group, SJHYR 2-treated group and normal saline (NS) control group. SJHYR 1 was prepared with Shengji Recipe (SJR, a compound traditional Chinese herbal medicine for promoting granulation) and Huayu Recipe (HYR, a compound traditional Chinese herbal medicine for resolving stagnation) at a ratio of 1:2, while SJHYR 2 was prepared with SJR and HYR at a ratio of 1:1. Immunohistochemical method was used to assess Bax and Bcl-2 protein levels in granulation tissue. RESULTS: SJHYR 1 could accelerate wound healing as compared with SJHYR 2 and NS (P<0.05). On the third day in experiment, Bax and Bcl-2 proteins were not found in any groups, but on the seventh and eleventh day in experiment, Bax and Bcl-2 proteins in SJHYR 1-treated group were much higher than those in the other two groups (P<0.05). CONCLUSION: SJR and HYR in different ratios may all have a role in regulating Bax and Bcl-2 expression in granulation tissue of diabetic rats during wound healing.

[Effects of Chinese herbs for replenishing qi and resolving stagnation on transforming growth factor-beta1 of skin ulcers in rats with diabetes].

OBJECTIVE: To explore the effects of Yiqi Huayu Recipe, a compound traditional Chinese herbal medicine for replenishing qi and resolving stagnation, on transforming growth factor-beta1(TGF-beta1) of skin ulcers in rats with diabetes. METHODS: Sixty rats were randomly divided into six groups, Yiqi Huayu Recipe-treated group, Yiqi (replenishing qi) Recipe-treated group, Huayu (resolving stagnation) Recipe-treated group, basic fibroblast growth factor (bFGF) treated group, untreated group and normal control group. Diabetes was induced by peritoneal injection of streptozotocin and skin ulcers were made by surgery method in rats except for the normal control group. Then the rats were administered with different drugs respectively, and the expression of TGF-beta1 in granulation tissue of the skin ulcers was detected with the methods of Western blotting, image analysis and immunohistochemistry. RESULTS: The level of TGF-beta1 expression in the untreated group was lower than that in the normal control group (P<0.01); and the level of TGF-beta1 expression in the drug-treated groups was higher than that in the untreated group (P<0.01); and the TGF-beta1 expression in the Yiqi Huayu Recipe-treated group was higher than that in the Yiqi Recipe-treated group, Huayu Recipe-treated group and bFGF-treated group (P<0.05). CONCLUSION: The replenishing qi and resolving stagnation therapy can control the secretion of TGF-beta1 of the wound in the process of wound healing in the levels of gene and molecule.

[Effects of Chinese herbs for replenishing qi and resolving stagnation on hypoxia-inducible factor-1alpha and vascular endothelial growth factor in granulation tissue of skin ulcers in rats with diabetes].

OBJECTIVE: To explore the effects of Chinese herbs for replenishing qi and resolving stagnation on hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in granulation tissue of skin ulcers in rats with syndrome of blood stasis and qi deficiency. METHODS: Diabetic rats with back full-thickness skin lesion and syndrome of blood stasis and qi deficiency were divided in to five groups: untreated group, basic fibroblast growth factor (bFGF)-treated group, Yiqi Huayu Recipe (a recipe for replenishing qi and resolving stagnation)-treated group, Yiqi Recipe (a recipe for replenishing qi)-treated group and Huayu Recipe (a recipe for resolving stagnation)-treated group, and another eight normal rats served as normal control group. Immunohistochemical method and image analysis were used to test the expressions of HIF-1alpha and VEGF in granulation tissue of skin ulcers in rats with diabetes. RESULTS: In the untreated group, the expression of HIF-1alpha was significantly increased and the expression of VEGF was significantly decreased as compared with those in the normal control group (P<0.01). The expression of HIF-1alpha was obviously lower (P<0.01) and the expression of VEGF was significantly higher in the four drug treated groups as compared with those in the untreated group. In the Yiqi Huayu Recipe-treated group, the expression of HIF-1alpha was obviously lower than those in the bFGF-treated, Yiqi Recipe-treated and Huayu Recipe-treated groups (P<0.05 or P<0.01), and the expression of VEGF was significantly higher than that in the bFGF-treated group (P<0.01). CONCLUSION: Chinese herbs for replenishing qi and resolving stagnation can promote the wound healing in rats through reducing the expression of HIF-1alpha, accelerating the expression of VEGF in granulation tissue of skin ulcers in rats with diabetes and ameliorating the status of ischemia and hypoxia.

Ischemic tissue oxygen capacitance after hyperbaric oxygen therapy: a new physiologic concept.

In an effort to better understand the effects of hyperbaric oxygen therapy on ischemic tissue, we monitored the real-time changes in subcutaneous tissue oxygen tension before, during, and after exposure to hyperbaric oxygen treatments. We identified an elevation of the tissue oxygen partial pressure to over 300 mmHg during the treatment period (up from a baseline mean of 24 mmHg) in a sustained ischemia rabbit ear model (n = 22 rabbits). There was no sustained change in tissue oxygen tension beyond the period of treatment. This manner of response is consistent with several current theories used to explain the mechanism of action of hyperbaric oxygen therapy. It is also consonant with our opinion that molecular oxygen, when delivered at high pressure, can function both as a respiratory metabolite and as a signal transducer. We also studied the impact of nontherapeutic 100% oxygen at 1 atm on tissue. The sustained peak tissue oxygen tension during such challenges increased in direct proportion to the number of hyperbaric oxygen treatments given. The clinical relevance and extension of these findings are discussed.