Objective:To observe the effects of moxibustion on colonic inflammation,and the expressions of ubiquitin and nucleotide-binding oligomerization domain(Nod)-Iike receptor protein 3(NLRP3)proteins in rats with ulcerativ...Objective:To observe the effects of moxibustion on colonic inflammation,and the expressions of ubiquitin and nucleotide-binding oligomerization domain(Nod)-Iike receptor protein 3(NLRP3)proteins in rats with ulcerative colitis(UC),and to explore the anti-inflammatory mechanism of moxibustion in the UC treatment.Methods:Clean grade male Sprague-Dawley(SD)rats were randomly divided into a normal group(NG),a model group(MG),a moxa-stick moxibustion group(MSMG)and a Western medicine group(WMG).UC model was prepared by freely drinking 35 g/L d ext ran sulfate sodium(DSS)solution.Bilateral Tianshu(ST 25)were selected for mild moxibustion treatment in the MSMG;mesalazine solution was intragastrically administrated in the WMG.Rats in the NG and MG were only grasped and fixed as in the MSMG without any treatment.After treatment,hematoxylin-eosin(HE)staining was performed to observe and score the colonic pathological damage under light microscope;immunofluorescence method was used to determine the expression of colonic ubiquitin protein;immunohistochemical method was used to detect the expressions of colonic interleukin(IL)-1β and NLRP3 proteins.Results:The colon tissue was severely injured,and the pathological score was significantly increased in the MG than in the NG(P<0.01),and the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon were significantly increased(all P<0.01).Compared with the MG,the colonic damage was repaired,the inflammation and pathological scores were reduced,and the ubiquitin,NLRP3 and IL-1β protein expressions were decreased in the MSMG and WMG(all P<0.01).Correlation analysis revealed that the ubiquitin protein expression was correlated with the colonic pathological score and the NLRP3 protein expression(r=0.677,P<0.01;r=0.536,P<0.05).Conclusion:Moxibustion can down-regulate the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon of UC rats,which may be one of the mechanisms to promote the repair of colonic inflammatory lesions and exert anti-inflammatory effects.展开更多
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...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.展开更多
Objective:To investigate the effect of electroacupuncture(EA)on cognitive function in D-galactose(D-gal)-induced aging rats,and the correlation between the effect and nucleotide-binding oligomerization domain(NOD)-lik...Objective:To investigate the effect of electroacupuncture(EA)on cognitive function in D-galactose(D-gal)-induced aging rats,and the correlation between the effect and nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)-ASC-Caspase-1 signaling pathway.Methods:Forty-six male Sprague-Dawley(SD)rats were randomly divided into a control group(n=10),a model group(n=12),an EA-7 d group(n=12)and an EA-21 d group(n=12).Except the control group,the other three groups received 42 consecutive days of intraperitoneal injection of D-gal to establish aging rat models with cognitive dysfunction.The control group received the same amount of normal saline via intraperitoneal injection.Two EA groups were given EA therapy for 21 consecutive days(began from the 22nd day of modeling)or 7 consecutive days(began from the 36th day of modeling)accordingly at Dazhui(GV 14),Baihui(GV 20),Shenshu(BL 23)and Zusanli(ST 36).After modeling/intervention,all four groups received behavioral evaluations by Morris water maze(MWM)test,novel object recognition(NOR)test and step-down passive avoidance(SDPA)test followed by the Western blot(WB)detection of the expression levels of hippocampal NLRP3 inflammasome-associated proteins NLRP3,ASC and Caspase-1.Results:MWM(place navigation test,PNT)results showed that the escape latency in the model group was significantly longer than that in the other three groups(P<0.05),and there was no significant difference among the other three groups on the 1st day of the test(P>0.05).From the 2nd day to the 4th day of the test,there was no significant difference between the EA-21 d group and the control group(P>0.05)in the escape latency;the escape latency was significantly shorter in the EA-21 d group than in the model group and the EA-7 d group(P<0.05).MWM(spatial probe test,SPT)results showed that the time spent in the target quadrant was significantly shorter and platform crossover number was significantly lower in the model group than in the other three groups(P<0.05).The time spent in the target quadrant was longer in the EA-7 d group than in the model group(P<0.05),but was shorter than that in the control group and the EA-21 d group(P<0.05).There was no significant difference in the swimming speed among the four groups(P>0.05).NOR results showed that there was no significant difference in the recognition ratio between the EA-7 d group and the EA-21 d group(P>0.05),and the recognition ratio was significantly higher in the two EA groups than in the model group(P<0.05),but was lower than in the control group(P<0.05).SDPA results showed that the electric shock number was higher in the model group than in the other three groups(P<0.05),and the differences among the other three groups were statistically insignificant(P>0.05).The model group had the shortest step-down latency,followed by the EA-7 d group,the EA-21 d group and the control group in order(P<0.05).The WB results indicated that the expression level of NLRP3 was significantly lower in the control group and the EA-21 d group than in the model group and the EA-7 d group(P<0.05).The expression levels of ASC and Caspase-1 were significantly higher in the model group than in the other three groups(P<0.05),and there was no significant difference among these three groups(P>0.05).Conclusion:NLRP3 inflammasome may be involved in the development of cognitive decline in aging rats;7 consecutive days of EA intervention can partially improve the cognitive impairment in aging rats though the effect is rather limited;21 consecutive days of EA intervention may improve the learning and memory abilities in aging rats via downregulating the expression levels of NLRP3 inflammasome-associated proteins in hippocampus.展开更多
炎症在众多疾病的发展中起着重要作用,而核苷酸结合寡聚化域样受体蛋白3(nucleotide binding oligomerization domain like receptors protein 3,NLRP3)信号通路是其中关键的一环。NLRP3可通过自组装后形成炎性小体激活半胱氨酸天冬氨...炎症在众多疾病的发展中起着重要作用,而核苷酸结合寡聚化域样受体蛋白3(nucleotide binding oligomerization domain like receptors protein 3,NLRP3)信号通路是其中关键的一环。NLRP3可通过自组装后形成炎性小体激活半胱氨酸天冬氨酸蛋白水解酶-1,进而激活IL-1β和IL-18前体引发炎症;而马来酸胺乙基青蒿素、骨化三醇、非诺贝特、MCC950、白藜芦醇等药物则可通过抑制NLRP3信号通路的不同位点(如促进核转录因子κB、半胱氨酸天冬氨酸蛋白水解酶-1、NLRP3等)的表达以降低IL-18和IL-1β的表达,进而实现控制氧化应激和慢性炎症进展,最终实现治疗干眼、糖尿病视网膜病变和AMD等眼病的目的。目前NLRP3信号通路抑制剂治疗眼病的相关研究尚停留在较为早期的动物模型及体外实验,其于眼科疾病的应用研究有待进一步拓展。展开更多
文摘Objective:To observe the effects of moxibustion on colonic inflammation,and the expressions of ubiquitin and nucleotide-binding oligomerization domain(Nod)-Iike receptor protein 3(NLRP3)proteins in rats with ulcerative colitis(UC),and to explore the anti-inflammatory mechanism of moxibustion in the UC treatment.Methods:Clean grade male Sprague-Dawley(SD)rats were randomly divided into a normal group(NG),a model group(MG),a moxa-stick moxibustion group(MSMG)and a Western medicine group(WMG).UC model was prepared by freely drinking 35 g/L d ext ran sulfate sodium(DSS)solution.Bilateral Tianshu(ST 25)were selected for mild moxibustion treatment in the MSMG;mesalazine solution was intragastrically administrated in the WMG.Rats in the NG and MG were only grasped and fixed as in the MSMG without any treatment.After treatment,hematoxylin-eosin(HE)staining was performed to observe and score the colonic pathological damage under light microscope;immunofluorescence method was used to determine the expression of colonic ubiquitin protein;immunohistochemical method was used to detect the expressions of colonic interleukin(IL)-1β and NLRP3 proteins.Results:The colon tissue was severely injured,and the pathological score was significantly increased in the MG than in the NG(P<0.01),and the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon were significantly increased(all P<0.01).Compared with the MG,the colonic damage was repaired,the inflammation and pathological scores were reduced,and the ubiquitin,NLRP3 and IL-1β protein expressions were decreased in the MSMG and WMG(all P<0.01).Correlation analysis revealed that the ubiquitin protein expression was correlated with the colonic pathological score and the NLRP3 protein expression(r=0.677,P<0.01;r=0.536,P<0.05).Conclusion:Moxibustion can down-regulate the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon of UC rats,which may be one of the mechanisms to promote the repair of colonic inflammatory lesions and exert anti-inflammatory effects.
文摘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.
文摘Objective:To investigate the effect of electroacupuncture(EA)on cognitive function in D-galactose(D-gal)-induced aging rats,and the correlation between the effect and nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)-ASC-Caspase-1 signaling pathway.Methods:Forty-six male Sprague-Dawley(SD)rats were randomly divided into a control group(n=10),a model group(n=12),an EA-7 d group(n=12)and an EA-21 d group(n=12).Except the control group,the other three groups received 42 consecutive days of intraperitoneal injection of D-gal to establish aging rat models with cognitive dysfunction.The control group received the same amount of normal saline via intraperitoneal injection.Two EA groups were given EA therapy for 21 consecutive days(began from the 22nd day of modeling)or 7 consecutive days(began from the 36th day of modeling)accordingly at Dazhui(GV 14),Baihui(GV 20),Shenshu(BL 23)and Zusanli(ST 36).After modeling/intervention,all four groups received behavioral evaluations by Morris water maze(MWM)test,novel object recognition(NOR)test and step-down passive avoidance(SDPA)test followed by the Western blot(WB)detection of the expression levels of hippocampal NLRP3 inflammasome-associated proteins NLRP3,ASC and Caspase-1.Results:MWM(place navigation test,PNT)results showed that the escape latency in the model group was significantly longer than that in the other three groups(P<0.05),and there was no significant difference among the other three groups on the 1st day of the test(P>0.05).From the 2nd day to the 4th day of the test,there was no significant difference between the EA-21 d group and the control group(P>0.05)in the escape latency;the escape latency was significantly shorter in the EA-21 d group than in the model group and the EA-7 d group(P<0.05).MWM(spatial probe test,SPT)results showed that the time spent in the target quadrant was significantly shorter and platform crossover number was significantly lower in the model group than in the other three groups(P<0.05).The time spent in the target quadrant was longer in the EA-7 d group than in the model group(P<0.05),but was shorter than that in the control group and the EA-21 d group(P<0.05).There was no significant difference in the swimming speed among the four groups(P>0.05).NOR results showed that there was no significant difference in the recognition ratio between the EA-7 d group and the EA-21 d group(P>0.05),and the recognition ratio was significantly higher in the two EA groups than in the model group(P<0.05),but was lower than in the control group(P<0.05).SDPA results showed that the electric shock number was higher in the model group than in the other three groups(P<0.05),and the differences among the other three groups were statistically insignificant(P>0.05).The model group had the shortest step-down latency,followed by the EA-7 d group,the EA-21 d group and the control group in order(P<0.05).The WB results indicated that the expression level of NLRP3 was significantly lower in the control group and the EA-21 d group than in the model group and the EA-7 d group(P<0.05).The expression levels of ASC and Caspase-1 were significantly higher in the model group than in the other three groups(P<0.05),and there was no significant difference among these three groups(P>0.05).Conclusion:NLRP3 inflammasome may be involved in the development of cognitive decline in aging rats;7 consecutive days of EA intervention can partially improve the cognitive impairment in aging rats though the effect is rather limited;21 consecutive days of EA intervention may improve the learning and memory abilities in aging rats via downregulating the expression levels of NLRP3 inflammasome-associated proteins in hippocampus.
文摘炎症在众多疾病的发展中起着重要作用,而核苷酸结合寡聚化域样受体蛋白3(nucleotide binding oligomerization domain like receptors protein 3,NLRP3)信号通路是其中关键的一环。NLRP3可通过自组装后形成炎性小体激活半胱氨酸天冬氨酸蛋白水解酶-1,进而激活IL-1β和IL-18前体引发炎症;而马来酸胺乙基青蒿素、骨化三醇、非诺贝特、MCC950、白藜芦醇等药物则可通过抑制NLRP3信号通路的不同位点(如促进核转录因子κB、半胱氨酸天冬氨酸蛋白水解酶-1、NLRP3等)的表达以降低IL-18和IL-1β的表达,进而实现控制氧化应激和慢性炎症进展,最终实现治疗干眼、糖尿病视网膜病变和AMD等眼病的目的。目前NLRP3信号通路抑制剂治疗眼病的相关研究尚停留在较为早期的动物模型及体外实验,其于眼科疾病的应用研究有待进一步拓展。
