RESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) and "Feishu" (BL 13) on pulmonary function, inflammatory reaction and expression of macrophage migration inhibitory factor (MIF) and its receptor complex CD 74-CD 44, etc. in rats with chronic obstructive pulmonary disease (COPD), so as to explore its mechanism underlying improvement of COPD. METHODS: Thirty male SD rats were randomly divided into normal, model and EA groups (nï¼10 in each group). The COPD model was established by intratracheal infusion of Lipopolysaccharide (LPS, 1 mg/mL) and forced smoke-inhaling. EA was applied to bilateral ST 36 and BL 13 for 30 min, once daily for 7 days. The rat's lung function (forced vital capacity [FVC], forced expiratory capacity ratio ([FEV 0.1/FVC] and [FEV 0.3/FVC]) was detected under anesthesia. Pathological changes of the lung tissue were detected by Hï¼E. staining, and the contents of MIF, tumor necrosis factor-α (TNF-α), interleukin-1 ß (IL-1 ß) and IL-8 in serum, bronchoalveolar lavage fluid (BALF) and lung tissue were assayed by ELISA. The immunoactivity of CD 74 and CD 44 was detected by immunohistochemistry, and the expression levels of MIF, CD 74, CD 44 and p 38 MAPK mRNAs and proteins were examined by quantitative RT-PCR and Western blot, respectively. RESULTS: Compared with the normal group, the FVC, FEV 0.1, FEV 0.3, FEV 0.1/FVC and FEV 0.3/FVC levels were significantly decreased in the model group (P<0.01). After EA treatment, the FVC, FEV 0.1, FEV 0.3, FEV 0.1/FVC and FEV 0.3/FVC were significantly increased (P<0.01, P<0.05), suggesting an improvement of the pulmonary function after EA. Hï¼E. staining showed that the severity of modeling induced alveolar expansion and inflammatory cell infiltration in the lung tissue was relatively milder in the EA group relevant to the model group. The contents of MIF, TNF-αï¼ IL-1 ß and IL-8 in the serum, BALF and lung tissues were significantly higher in the model group than in the normal group (P<0.01), and significantly down-regulated in the EA group relevant to the model group (P<0.01). The expression levels of MIF, CD 74, CD 44 and p 38 MAPK mRNAs and proteins and the immunoactivity levels of CD 74, CD 44 in the lung tissue were obviously higher in the model group than those in the normal group (P<0.01), and considerably lower in the EA group than those in the model group (P<0.01). There was a positive correlation between p 38 MAPK and MIF in mRNA and protein expression levels (P<0.01). CONCLUSION: EA intervention can improve the pulmonary function in COPD rats, which may be related to its effects in inhibiting inflammatory reaction, and MIF/CD 74-CD 44/p 38 MAPK signaling pathway.
Assuntos
Eletroacupuntura , Doença Pulmonar Obstrutiva Crônica , Animais , Antígenos de Diferenciação de Linfócitos B , Antígenos de Histocompatibilidade Classe II , Receptores de Hialuronatos , Pulmão , Sistema de Sinalização das MAP Quinases , Fatores Inibidores da Migração de Macrófagos , Masculino , Doença Pulmonar Obstrutiva Crônica/terapia , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: Acupuncture has a definite therapeutic effect on chronic obstructive pulmonary disease (COPD), and the cholinergic anti-inflammatory pathway (CAP) has been shown to be involved in regulation of inflammation. In this study, we investigated whether electro-acupuncture (EA) affects the CAP in COPD. METHODS: Sprague-Dawley rats were induced into COPD through exposure to cigarette smoke combined with lipopolysaccharide. EA treatment was applied at Zusanli (ST36) and Feishu (BL13) points for 30â¯min/d for 7â¯d. Seventy-two rats were randomly divided into six study groups, including normal, normalâ¯+â¯EA, normalâ¯+â¯α-bungarotoxin (α-BGT) (the antagonist of the nicotinic acetylcholine receptor α7 subunit (α7nAChR))â¯+â¯EA, COPD, COPDâ¯+â¯EA, and COPDâ¯+â¯α-BGTâ¯+â¯EA. Lung function, pathology and vagus nerve discharge were tested. The levels of acetylcholine (ACh), acetylcholinesterase (AChE), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in bronchoalveolar lavage fluid (BALF) and lung tissue were measured by enzyme-linked immunosorbent assay. The mRNA and protein expression and immunoreactivity of α7nAChR and its postreceptor inflammation signal pathway, including janus kinase 2 (JAK2), signal transducers and activators of transcription 3 (STAT3), nuclear factor-κB (NF-κB), were observed by quantitative reverse transcription-polymerase chain reaction, Western blot and immunohistochemistry. RESULTS: Compared with normal rats, there were a significant decline in lung function and discharge of the vagus nerve (Pâ¯<â¯0.01), a marked sign of lung inflammation and an increase of ACh, AChE, IL-6 and TNF-α level in BALF or lung tissue (Pâ¯<â¯0.05, Pâ¯<â¯0.01) and higher expression of α7nAChR, JAK2, STAT3 and NF-κB (Pâ¯<â¯0.05, Pâ¯<â¯0.01) in the COPD rats. In rats receiving EA, the lung function and vagal discharge were enhanced (Pâ¯<â¯0.01), lung inflammation was improved and the levels of ACh, AChE, IL-6 and TNF-α were decreased (Pâ¯<â¯0.01). Further, the expression of α7nAChR, JAK2, STAT3 and NF-κB was downregulated (Pâ¯<â¯0.05, Pâ¯<â¯0.01). However, the above effects of EA were blocked in rats injected with α-BGT (Pâ¯<â¯0.01). CONCLUSION: EA treatment can reduce the lung inflammatory response and improve lung function in COPD, which may be related to its involvement in the regulation of CAP.
Assuntos
Acetilcolina/imunologia , Eletroacupuntura , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/terapia , Animais , Modelos Animais de Doenças , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Pulmão/imunologia , Masculino , NF-kappa B/genética , NF-kappa B/imunologia , Doença Pulmonar Obstrutiva Crônica/genética , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
The aim of the present study was to investigate the influence and the mechanisms of cineole and terpineol on the in-vitro transdermal delivery of huperzine A from microemulsions, and their potential synergistic effect on the permeation enhancement. The transdermal delivery of huperzine A from microemulsions with different concentrations of cineole and terpineol through the rat abdominal skin was determined by Franz-type diffusion cells. The partition coefficient of huperzine A between the full thickness skin and microemulsion was determined. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) was carried out to analyze the effects of cineole and terpineol on the biophysical properties of the stratum corneum (SC) and the mechanisms of permeation enhancement. These results indicated that cineole and terpineol could synergistically increase the transdermal delivery of huperzine A from microemulsions through increasing the partition and diffusion coefficients of huperzine A. ATR-FTIR studies further validated the synergistic effect and revealed that the enhancing mechanisms were due to increasing the disorderliness and fluidity of SC lipid alkyl chains, disrupting the structure of keratin in SC, and extracting SC lipids. In conclusion, cineole and terpineol, acting synergistically to enhance the transdermal delivery of huperzine A from microemulsions, might provide an alternative permeation enhancer combination for the transdermal delivery of huperzine A.
RESUMO
Shaoyao-Gancao decoction (SGD), a traditional Chinese formulation containing Paeoniae Radix (SY) and Glycyrrhizae Radix (GC), is commonly used to relieve abdominal pain. However, the absorption and metabolites of the characteristic constituents of the two herbs in vivo have been reported rarely. The purpose of this study was to investigate the compatibility rationality and the mechanism of the enhanced efficiency of SGD. A single or a mixed decoction (SG and S+G, respectively) was orally administered to rats. Blood samples were collected at different intervals following treatment and analyzed by liquid LC/MS. A total of fifteen ingredients (denoted as M1 to M15) were found in both rat plasma after treatment with the two decoctions. Furthermore, the proposed structures of the remained twelve ingredients were obtained except M9, M10 and M15. The quality of the ingredients in the rat plasma showed no significant difference between the two decoctions. However, the quantity of twelve ingredients differed greatly, indicating that the absorption of SG was greater than that of S+G except M7, M12 and M15. As the compositions associated with the efficacy of SG and S+G were inconsistent, the degree of the absorption of the 15 ingredients by the gastrointestinal tract were different, which caused a significantly enhanced efficacy of certain ingredients. This study presents an exploration of the mechanism behind the improved efficacy of individual components in traditional Chinese medicine therapies through combination with other components.
Assuntos
Análise Química do Sangue , Cromatografia Líquida , Medicamentos de Ervas Chinesas , Glycyrrhiza/química , Espectrometria de Massas , Paeonia/química , Absorção , Administração Oral , Animais , Glycyrrhiza/metabolismo , Masculino , Estrutura Molecular , Paeonia/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Shaoyao-Gancao decoction, a traditional Chinese formulation composed of Paeoniae Radix and Glycyrrhizae Radix, is commonly used to relieve abdominal pain. In this paper, the compatibility rationality of this decoction was investigated. Shaoyao-Gancao decoction, Shaoyao decoction and Gancao decoction were orally administered to rats, respectively. Blood samples were collected at pre-determined times after administration and analyzed by high-performance liquid chromatography (HPLC). The pharmacokinetic parameters of characteristic peaks were analyzed and the statistical significance of the obtained parameters was determined. Paeoniflorin (12.0 min) and compounds at retention times of 4.7 and 5.2 min were all significantly higher in the Shaoyao-Gancao decoction than in the Shaoyao decoction (P < 0.05). In contrast, in the Gancao decoction, the compound at a retention time of 14.6 min was significantly lower than in the Shaoyao-Gancao decoction (P < 0.01). However, the compounds at retention times of 17.1 and 18.1 min were significantly higher in the Gancao decoction than in the Shaoyao-Gancao decoction (P < 0.05). These results indicate that poor compatibility of the compounds in the Shaoyao-Gancao decoction could result in poor absorption. The compatibility of the component compounds of the Shaoyao-Gancao decoction was revealed in the pharmacokinetic characteristics of the decoction. Generally, the absorption of Shaoyao components was increased in the Shaoyao-Gancao decoction, while the absorption of Gancao components was time dependent. In the Shaoyao-Gancao decoction, the increased absorption of some Shaoyao components may be related to a reduction in absorption of some Gancao components.