Mechanism of Qishen Decoction inhibition of macrophage M1 type polarization by targeting TGR5-mediated NLRP3 inflammasome

Objective:To observe the effect of Qishen decoction on TGR5-mediated activation of NLRP3 inflammasome,so as to clarify the molecular mechanism of its inhibition of macrophage M1-type polarisation to ameliorate non-alcoholic steatohepatitis;Methods:Mouse macrophage cell line RAW264.7 was randomly divided into a control group,model group,Qishen decoction group,TGR5 agonist group and Qishen decoction+TGR5 agonist group.Except for the control group,the remaining groups were constructed the macrophage NLRP3 activation model by palmitic acid induction,and the corresponding drugs were given to intervene.ELISA was used to detect the levels of TNF-α,IL-6,IL-1βand CXCL2 in macrophage supernatants,flow cytometry was used to detect the expression levels of macrophage polarisation marker molecules CD86 and iNOS,and Western blot was used to detect the expression of the TGR5/STAT1/STAT6 signaling pathway and the expression of NLRP3 inflammasome-associated proteins,respectively.Results:Compared with the control group,the contents of macrophages TNF-α,IL-6,IL-1β,CXCL2 and the proportion of macrophages with positive expression of CD86 and iNOS were significantly increased in the model group,and the differences were all statistically significant(P<0.01).Compared with the model group,the contents of TNF-α,IL-6,IL-1β,CXCL2 and the proportion of macrophages with positive expression of CD86 and iNOS were significantly decreased in the Qishen decoction group,and the differences were all statistically significant(P<0.01).In addition,the expression of NLRP3 and Pro-IL-1βproteins in the macrophage lysate and the expression of Caspase-1 p10,Caspase-1 p20 and IL-1βp17 proteins in the cell supernatant of the model group were significantly increased when compared with the control group,and the differences were all statistically significant(P<0.01).Compared with the model group,the expression of NLRP3 and Pro-IL-1βproteins in macrophage lysate and the expression of Caspase-1 p10,Caspase-1 p20 and IL-1βp17 proteins in cell supernatant of the Qishen decoction were significantly reduced,and the differences were all statistically significant(P<0.01);Conclusion:Qishen decoction can inhibit the activation of NLRP3 inflammasome in macrophages by inhibiting the TGR5/STAT1/STAT6 signaling pathway,thereby inhibiting macrophage M1 polarization and improving inflammatory response.

Acupuncture Treatment of Habitual Constipation

The authors have in recent years used acupuncture to treat 188 cases of habitual constipation with remarkable curative effects. A report follows.

针刺对单眼剥夺大鼠视皮层NMDAR1 mRNA表达调节的研究（英文）

目的:探讨针刺干预视觉剥夺效应的分子生物学机制。方法:采用随机数字表法将48只2周龄的Wistar大鼠随机分为正常组、模型组和6个针刺组[早期针刺患侧组(C1)、早期针刺健侧组(C2)、中期针刺患侧组(D1)、中期针刺健侧组(D2)、晚期针刺患侧组(E1)和晚期针刺健侧组(E2)],每组6只。正常组不予任何处理;模型组和各针刺组采用单侧眼睑缝合的方法建立剥夺性弱视动物模型,造模成功后,模型组不予任何治疗;早期、中期、晚期针刺组分别于造模后第3天、12天、21天开始针刺治疗。治疗结束后检测各组大鼠图形视觉诱发电位(P-VEP)和视皮层17区N-甲基-D-天门冬氨酸受体1(NMDAR1)m RNA的表达。结果:模型组大鼠P-VEP波形较正常组有明显改变,表现为P_(100)时值明显延迟(P<0.01),N_(45)-P_100幅值显著降低(P<0.01);治疗后,各针刺组大鼠P-VEP波形较模型组均有显著改善,表现为P_(100)时值明显提前(P<0.01),N45-P_(100)幅值明显升高(P<0.05);并且早期针刺对P-VEP波形的影响大于中期和晚期针刺。模型组大鼠视皮层17区NMDAR1 mRNA的表达较正常组明显降低(P<0.01);而治疗后各针刺组大鼠视皮层17区NMDAR1 m RNA表达较模型组均显著提高(P<0.05);其中早期针刺对NMDAR1 mRNA表达水平的影响大于中期和晚期针刺;而早期针刺患侧穴位对NMDAR1 mRNA表达水平的影响优于针刺健侧穴位(P<0.05);中期、晚期针刺患侧穴位与健侧穴位对NMDAR1 mRNA表达水平的影响差异无统计学意义(P>0.05)。结论:单眼剥夺后大鼠P-VEP波形出现异常改变,视皮层17区NMDAR1 mRNA表达减少。敏感期内针刺对单眼剥夺大鼠异常的P-VEP波形和视皮层17区NMDAR1 mRNA低水平表达均具有明显的调节作用,并且敏感期内早期治疗是取得疗效的关键。