RESUMO
OBJECTIVE@#To explore whether the ethanol extract of Herpetospermum caudigerum Wall (EHC), a Xizang medicinal plant traditionally used for treating liver diseases, can improve imiquimod-induced psoriasis-like skin inflammation.@*METHODS@#Immunohistochemistry and immunofluorescence staining were used to determine the effects of topical EHC use in vivo on the skin pathology of imiquimod-induced psoriasis in mice. The protein levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-17A (IL-17A) in mouse skin samples were examined using immunohistochemical staining. In vitro, IFN-γ-induced HaCaT cells with or without EHC treatment were used to evaluate the expression of keratinocyte-derived intercellular cell adhesion molecule-1 (ICAM-1) and chemokine CXC ligand 9 (CXCL9) using Western blotting and reverse transcription-quantitative polymerase chain reaction. The protein synthesis inhibitor cycloheximide and proteasome inhibitor MG132 were utilized to validate the EHC-mediated mechanism underlying degradation of ICAM-1 and CXCL9.@*RESULTS@#EHC improved inflammation in the imiquimod-induced psoriasis mouse model and reduced the levels of IFN-γ, TNF-α, and IL-17A in psoriatic lesions. Treatment with EHC also suppressed ICAM-1 and CXCL9 in epidermal keratinocytes. Further mechanistic studies revealed that EHC suppressed keratinocyte-derived ICAM-1 and CXCL9 by promoting ubiquitin-proteasome-mediated protein degradation rather than transcriptional repression. Seven primary compounds including ehletianol C, dehydrodiconiferyl alcohol, herpetrione, herpetin, herpetotriol, herpetetrone and herpetetrol were identified from the EHC using ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry.@*CONCLUSION@#Topical application of EHC ameliorates psoriasis-like skin symptoms and improves the inflammation at the lesion sites. Please cite this article as: Zhong Y, Zhang BW, Li JT, Zeng X, Pei JX, Zhang YM, Yang YX, Li FL, Deng Y, Zhao Q. Ethanol extract of Herpetospermum caudigerum Wall ameliorates psoriasis-like skin inflammation and promotes degradation of keratinocyte-derived ICAM-1 and CXCL9. J Integr Med. 2023; 21(6): 584-592.
Assuntos
Animais , Camundongos , Interleucina-17/metabolismo , Molécula 1 de Adesão Intercelular , Imiquimode/efeitos adversos , Fator de Necrose Tumoral alfa/metabolismo , Ligantes , Psoríase/induzido quimicamente , Queratinócitos , Inflamação/tratamento farmacológico , Quimiocinas/metabolismo , Interferon gama/metabolismo , Modelos Animais de Doenças , Camundongos Endogâmicos BALB CRESUMO
OBJECTIVE@#Psoriasis is a common chronic inflammatory skin disease that is prone to recurrence, and the proinflammatory factor, cysteine-rich protein 61 (Cyr61), is important in its pathophysiology. Long-term clinical practice has shown that Sancao Formula (SC), a Chinese herbal compound, is effective in the treatment of psoriasis, but the precise mechanism remains unknown. In this study, we investigate the mechanism by which SC extract alleviates imiquimod (IMQ)-induced psoriasis.@*METHODS@#The expression of Cyr61 in psoriatic lesions and normal healthy skin was detected using immunohistochemical analysis to investigate the biological role of Cyr61 in models of psoriatic inflammation. A psoriatic mouse model was established by topical application of IMQ, and the effect of topical application of SC extract was evaluated using the psoriasis area and severity index (PASI) score, hematoxylin-eosin staining, and histopathological features of the skin. Next, a HaCaT cell inflammation model was established using interferon-γ (IFN-γ), and the effect of SC extract on the mRNA and protein levels of Cyr61 and intercellular cell adhesion molecule-1 (ICAM-1) was confirmed using Western blot and quantitative real-time polymerase chain reaction analyses.@*RESULTS@#Immunohistochemical staining showed that the expression of Cyr61 in psoriatic lesions was higher than that in normal skin samples (78.26% vs 41.18%, P < 0.05), and the number of Cyr61-positive cells in psoriatic lesions was also significantly higher than in normal skin (18.66 ± 2.51 vs 4.33 ± 1.52, P < 0.05). Treatment in mice with IMQ-induced psoriasis showed that SC extract could significantly improve the inflammatory phenotype, PASI score (10.875 ± 0.744 vs 3.875 ± 0.582, P < 0.05), and pathological features compared with those in IMQ model group; SC treatment was also associated with decreased levels of Cyr61 and ICAM-1. In the IFN-γ-induced inflammatory cell model, the mRNA and protein levels of Cyr61 and ICAM-1 were upregulated, while the SC extract downregulated the levels of Cyr61 and ICAM-1.@*CONCLUSION@#The results provide a theoretical basis for the involvement of Cyr61 in the pathogenesis of psoriasis, and suggest that SC should be used to target Cyr61 for the prevention of psoriasis recurrence.
Assuntos
Animais , Camundongos , China , Proteína Rica em Cisteína 61/metabolismo , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/uso terapêutico , Imiquimode/efeitos adversos , Inflamação/tratamento farmacológico , Molécula 1 de Adesão Intercelular/genética , Interferon gama , Camundongos Endogâmicos BALB C , Psoríase/patologia , RNA Mensageiro/uso terapêuticoRESUMO
<p><b>OBJECTIVE</b>To investigate the levels of cytokines related to T-helper (Th) 17 cells in serum and signal transducers in the psoriatic lesions of patients with psoriasis vulgaris of blood-heat syndrome (BHS) and blood-stasis syndrome (BSS).</p><p><b>METHODS</b>Sixty patients with psoriasis vulgaris were divided into the BHS and BSS groups according to the syndrome differentiation of Chinese medicine (CM). Ten healthy subjects were considered as the control group. Cytokine levels of interleukin (IL)-17, IL-23 and IL-6 in serum were determined by enzyme-linked immunosorbent assay. Expression levels of signal transducer and activator of transcription 3 (STAT3), p38-mitogen-activated protein kinase (MAPK) and STAT6 in the psoriatic lesions were determined using immunohistochemistry (IHC), Western blot, and real-time quantitative reverse transcription polymerase chain reaction, respectively.</p><p><b>RESULTS</b>Production of IL-17, IL-23 and IL-6 in the BHS group and BSS group were significantly increased compared with those in the control group (P<0.05). Levels of IL-17 and IL-23 in the BHS group were higher than those in the BSS group (P<0.05). Compared with the control group, IHC positive expressions and protein expressions of STAT3 and p38-MAPK, and the STAT3 mRNA expressions in the BHS and BSS groups were significantly higher (P<0.05 or P<0.01). The protein expression of STAT3 in the BHS group was significantly higher than that in the BSS group (P<0.05).</p><p><b>CONCLUSIONS</b>Cytokines in serum and signal transducers in the psoriatic lesions alter with various CM syndromes of psoriasis. The results provide scientific basis for the treatment based on syndrome differentiation of CM in treating psoriasis vulgaris.</p>
Assuntos
Adulto , Feminino , Humanos , Masculino , Regulação da Expressão Gênica , Imuno-Histoquímica , Interleucina-17 , Sangue , Interleucina-23 , Sangue , Interleucina-6 , Sangue , Psoríase , Sangue , Genética , Alergia e Imunologia , RNA Mensageiro , Genética , Metabolismo , Fator de Transcrição STAT3 , Genética , Metabolismo , Fator de Transcrição STAT6 , Genética , Metabolismo , Transdução de Sinais , Síndrome , Células Th17 , Alergia e Imunologia , Proteínas Quinases p38 Ativadas por Mitógeno , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To explore the correlation between syndrome types of late-onset acne female patients and constitutions of Chinese medicine (CM).</p><p><b>METHODS</b>A questionnaire was performed in 365 late-onset acne female patients and 135 healthy subjects (as the control) using Professor WANG Qi's. methods and Standards for Chinese Medical Constitutions Classification.</p><p><b>RESULTS</b>Their CM constitutions were sequenced as damp-heat constitution, yin-deficiency constitution, balanced constitution, yang-deficiency constitution, blood-stasis constitution, qi-stagnation constitution, qi-deficiency constitution, phlegm-damp constitution, inherited special constitution, with statistical difference when compared with those of the control group ( χ2 = 85.206, P < 0.01). In the 365 female late-onset acne patients, 114 (31.23%) were with Chongren imbalance syndrome, 108 (29.59%) were with blood stasis or coagulated phlegm syndrome, 83 (22.74%) were with dampness heat syndrome, and 60 (16.44%) were with wind heat syndrome. There was statistical difference in CM constitution distributions among different CM syndrome types (χ2 = 105.671, P < 0.01). The distribution of CM medical constitutions was different between the two groups. Biased constitutions were often seen in the patient group, while balanced constitution was often seen in the control group. Binary Logistic regression analysis indicated that influencing factors covered sweet food, light diet, roasted food, coffee, stress, work pressure, and family pressure. Of them light diet was one protective factor, while the rest were adverse factors.</p><p><b>CONCLUSION</b>The etiology and syndrome types of female late-onset acne female patients were associated with CM constitution.</p>
Assuntos
Feminino , Humanos , Acne Vulgar , Epidemiologia , Constituição Corporal , Medicina Tradicional Chinesa , Inquéritos e Questionários , Síndrome , Deficiência da Energia Yang , Deficiência da Energia YinRESUMO
<p><b>OBJECTIVE</b>To explore the correlation between the pathogenesis of psoriasis patients of blood heat syndrome (BHS) and blood stasis syndrome (BSS) and peripheral blood Th1/Th2 cells axis drift, and to observe different expressions of peripheral blood Th1/Th2 cells between healthy subjects and psoriasis patients of BHS and BSS.</p><p><b>METHODS</b>There were 15 patients in the BHS group and 15 in the BSS group. There were 16 patients in the healthy control group. The expressions of CD4+ gamma-interferon (IFN-gamma) and interleukin-4 (IL-4) in the peripheral blood mononuclear cells (PBMCs) were measured by flow cytometry (FACS). The Th1 main cytokines such as IFN-gamma and Th2 cytokines such as IL-4 in the serum of psoriasis patients of different syndromes were detected by enzyme-linked immunosorbent assay (ELISA). The Psoriasis Area and Severity Index score (PASI) were conducted.</p><p><b>RESULTS</b>FACS results showed that the expression level of CD4+ IFN-gamma+ in the PBMCs was significantly higher in the BHS group than in the BSS group and the healthy control group (P < 0.05). Besides, it was positively correlated with the PASI (P < 0.05). ELISA results showed that the peripheral serum level of IFN-gamma was significantly higher in the BHS group than in the BSS group and the healthy control group (P < 0.05). The plasma level of IFN-gamma was positively correlated with the PASI score in the BHS group (P < 0.05). The plasma level of IFN-gamma was negatively correlated with the PASI score in the BSS group (P < 0.05). The peripheral serum level of IL-4 was significantly lower in the BHS group than in the BSS group and the healthy control group (P < 0.05).</p><p><b>CONCLUSIONS</b>Peripheral Th1 cells had dominant state in psoriasis patients of BHS. When psoriasis patients of BHS were transformed to BSS or to the normal level, the expression of peripheral blood Th1 cells decreased.</p>
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Estudos de Casos e Controles , Interferon gama , Sangue , Interleucina-4 , Sangue , Medicina Tradicional Chinesa , Psoríase , Sangue , Diagnóstico , Células Th1 , Metabolismo , Equilíbrio Th1-Th2 , Células Th2 , MetabolismoRESUMO
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>
Assuntos
Animais , Masculino , Ratos , Diabetes Mellitus Experimental , Metabolismo , Medicamentos de Ervas Chinesas , Farmacologia , Usos Terapêuticos , Metaloproteinase 3 da Matriz , Metabolismo , Fitoterapia , Ratos Sprague-Dawley , Pele , Patologia , Úlcera Cutânea , Tratamento Farmacológico , Metabolismo , Inibidor Tecidual de Metaloproteinase-1 , MetabolismoRESUMO
<p><b>OBJECTIVE</b>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 β-catenin, c-myc and K6.</p><p><b>METHODS</b>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 β-catenin, c-myc, and K6.</p><p><b>RESULTS</b>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 β-catenin, c-myc and K6 compared with the normal skin. β-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 β-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.</p><p><b>CONCLUSIONS</b>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.</p>
Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações do Diabetes , Metabolismo , Terapêutica , Imuno-Histoquímica , Medicina Tradicional Chinesa , Polissacarídeos Bacterianos , Metabolismo , Estudos Prospectivos , Proteínas Proto-Oncogênicas c-myc , Metabolismo , Método Simples-Cego , Úlcera Cutânea , Metabolismo , Terapêutica , beta Catenina , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To explore the effect of apigenin on semen parameters in male mice.</p><p><b>METHODS</b>Totally 100 healthy male mice of Kunming strain were randomly divided into 5 groups according to the body weight: negative control, solvent control, low-dose apigenin, median-dose apigenin and high-dose apigenin, the latter three groups given intragastric apigenin at a fixed time every day for 7 and 14 days. At 35 days after the first medication, all the mice were killed and detected for the sperm motion parameters by computer aided sperm analysis (CASA).</p><p><b>RESULTS</b>There were no statistically significant differences in sperm motion parameters, density and motility between the negative control and the three apigenin groups after 7-day medication. At 14 days, the high-dose apigenin group showed remarkable decreases in average path velocity (VAP), straight line velocity (VSL), straightness (STR), wobbliness (WOB), the percentage of grade b sperm and sperm motility, and a significant increase in beat cross frequency (BCF) as compared with the negative control group (P < 0.05).</p><p><b>CONCLUSION</b>Apigenin affects sperm motility in male mice to a certain extent.</p>