ABSTRACT
Objective: To observe the effect of electroacupuncture (EA) on nuclear factor kappa B (NF-κB) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome in uterine tissues of rats with primary dysmenorrhea (PD), thus to explore the possible mechanism of EA for PD. Methods: Fifty female Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, an EA at non-acupoint group, an EA at acupoint group and a Western medicine group, with 10 rats in each group. Except for the normal group, rats in the other four groups were treated with estradiol benzoate combined with oxytocin for 11 d to establish PD rat models. From day 1 of the modeling, rats in the normal group and the model group were only properly grasped without any intervention; Guanyuan (CV 4) and Sanyinjiao (SP 6) were selected for EA treatment in the EA at acupoint group; rats in the EA at non-acupoint group were treated with EA at 5 mm away from the acupoints selected above; rats in the Western medicine group were treated with ibuprofen via gavage. Rats in each group were treated for 10-day successively. On the 11th day, except for the normal group, rats in the other groups were intraperitoneally injected with oxytocin (2 U/rat), and the writhing number within 30 min in each group was compared; the pathological changes in rat uteruses were observed by hematoxylin-eosin (HE) staining, and the pathological damage scores were evaluated. Protein expression levels of NF-κB p65, phospho-NF-κB p65, NLRP3, cysteine aspastic acid-specific protease 1 (caspase-1), interleukin (IL)-1β and IL-18 were detected by Western blot. Results: Compared with the normal group, the writhing number increased significantly (P<0.05), and the extensive exfoliation of the endometrium, severe edema, and histopathological score all increased significantly in the model group (P<0.05) as well as the protein levels of NLRP3, caspase-1, IL-1β and IL-18, and the ratio of phospho-NF-κB p65/NF-κB p65 in rat uterine tissues (all P<0.05); compared with the model group, the numbers of writhing reaction decreased within 30 min (P<0.05), the endometrial exfoliation was rare, the edema degree was mild, and the histopathological scores decreased significantly (all P<0.05) in the EA at acupoint group and the Western medicine group; compared with the model group, the phospho-NF-κB p65/NF-κB p65 ratio and the NLRP3, caspase-1, IL-1β and IL-18 protein levels of rat uterine tissues in the EA at acupoint group were significantly lower (P<0.05); compared with the model group, the caspase-1, IL-1β and IL-18 protein levels of the rat uterine tissues decreased significantly (all P<0.05), and the differences in the NLRP3 and phospho-NF-κB p65/NF-κB p65 levels were statistically insignificant (all P>0.05) in the Western medicine group; compared with the Western medicine group, the phospho-NF-κB p65/NF-κB p65 ratio, also the NLRP3, IL-1β and IL-18 protein levels of the uterine tissues decreased significantly in the EA at acupoint group (all P<0.05), while the difference in the caspase-1 level was statistically insignificant (P>0.05); there were no significant differences between the EA at non-acupoint group and the model group in any indicators (all P>0.05). Conclusion: EA at acupoints significantly improves the pain and pathological damages of PD rats. The mechanism may be related to the reduced uterine inflammation via inhibiting NF-κB phosphorylation and NLRP3 activation in uteruses of PD rats.
ABSTRACT
AIM:To investigate the effects of simvastatin on the expression of Toll-like receptor 2 ( TLR-2 ) , interferon-γ(IFN-γ) and monocyte chemoattractant protein-1 (MCP-1) in lung tissues of mice with mouse cytomegalovirus ( MCMV) pneumonia and to explore the possible mechanism .METHODS:Male BALB/c mice (6~8 weeks old, n=40) were randomly divided into 5 groups: normal control (NC) group, MCMV infection group, simvastatin group 1 (SMV1 group), simvastatin group 2 (SMV2 group), and simvastatin group 3 (SMV3 group).The mice in SMV1, SMV2 and SMV3 groups were gavaged with simvastatin (50 mg· kg-1 · d-1 for 7 d) 7 d before, on the same day of and 3 d after in-traperitoneal injection of MCMV , while the mice in normal control group and MCMV infection group were gavaged with the same volume of normal saline .HE staining was used to observe the pathological changes of lung tissues in mice .Total tis-sue protein was extracted from the lung homogenates to detect the expression of TLR-2 by Western blot and immunohisto-chemical staining .Real-time PCR was used to analyse the content of MCMV DNA .The levels of IFN-γand MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA).RESULTS:Compared with NC group, the pathological chan-ges of the lung tissues of the mice in MCMV group showed alveolar interstitial edema , alveolar wall widening and a large number of inflammatory cells .The expression of TLR-2 in the lung tissues of the mice in model group was increased signifi-cantly.The content of MCMV DNA was increased , and the expression of IFN-γand MCP-1 was also increased significant-ly.Compared with the mice in MCMV group , the pathological changes of the lung tissues of simvastatin groups showed that the inflammatory cells were decreased .The expression of TLR-2 was down-regulated.The content of MCMV DNA was de-creased, and the levels of IFN-γand MCP-1 were also decreased significantly .At the same time, the expression of TLR-2 and the content of MCMV DNA in SMV1 group were less than those in SMV2 and SMV3 groups (P<0.05), and no statis-tically significant difference between SMV 2 and SMV3 groups was observed .CONCLUSION:Simvastatin down-regulates the TLR-2 signaling pathway , and reduces the expression of TLR-2 and replication of MCMV DNA , thus attenuating the pathological damage of the lung tissue .Early intervention with simvastatin plays an important role in preventing the infection of MCMV and reducing the inflammation .
ABSTRACT
AIM:To investigate the effects of simvastatin on the expression of Toll-like receptor 2 ( TLR-2 ) , interferon-γ(IFN-γ) and monocyte chemoattractant protein-1 (MCP-1) in lung tissues of mice with mouse cytomegalovirus ( MCMV) pneumonia and to explore the possible mechanism .METHODS:Male BALB/c mice (6~8 weeks old, n=40) were randomly divided into 5 groups: normal control (NC) group, MCMV infection group, simvastatin group 1 (SMV1 group), simvastatin group 2 (SMV2 group), and simvastatin group 3 (SMV3 group).The mice in SMV1, SMV2 and SMV3 groups were gavaged with simvastatin (50 mg· kg-1 · d-1 for 7 d) 7 d before, on the same day of and 3 d after in-traperitoneal injection of MCMV , while the mice in normal control group and MCMV infection group were gavaged with the same volume of normal saline .HE staining was used to observe the pathological changes of lung tissues in mice .Total tis-sue protein was extracted from the lung homogenates to detect the expression of TLR-2 by Western blot and immunohisto-chemical staining .Real-time PCR was used to analyse the content of MCMV DNA .The levels of IFN-γand MCP-1 were measured by enzyme-linked immunosorbent assay (ELISA).RESULTS:Compared with NC group, the pathological chan-ges of the lung tissues of the mice in MCMV group showed alveolar interstitial edema , alveolar wall widening and a large number of inflammatory cells .The expression of TLR-2 in the lung tissues of the mice in model group was increased signifi-cantly.The content of MCMV DNA was increased , and the expression of IFN-γand MCP-1 was also increased significant-ly.Compared with the mice in MCMV group , the pathological changes of the lung tissues of simvastatin groups showed that the inflammatory cells were decreased .The expression of TLR-2 was down-regulated.The content of MCMV DNA was de-creased, and the levels of IFN-γand MCP-1 were also decreased significantly .At the same time, the expression of TLR-2 and the content of MCMV DNA in SMV1 group were less than those in SMV2 and SMV3 groups (P<0.05), and no statis-tically significant difference between SMV 2 and SMV3 groups was observed .CONCLUSION:Simvastatin down-regulates the TLR-2 signaling pathway , and reduces the expression of TLR-2 and replication of MCMV DNA , thus attenuating the pathological damage of the lung tissue .Early intervention with simvastatin plays an important role in preventing the infection of MCMV and reducing the inflammation .