Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in ...Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in the method were from the Physical Examination Center of Anhui Provincial Hospital of Traditional Chinese Medicine,and the 60 rheumatoid arthritis patients included were from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine.The disease activity score of patients with rheumatoid arthritis was evaluated using VAS score and DAS28 score,joint symptoms and spleen deficiency syndrome score were evaluated using graded quantification method,immune inflammation indicators were detected using relevant instruments,inflammatory cytokines were detected using ELISA method,macrophage markers were detected using FCM method,and pathway gene expression was detected using RT-qPCR;Evaluate the predictive effect of circ-CBLB and miR-486-5p on disease activity in rheumatoid arthritis using ROC curves.Results:(1)miR-486-5p,CD14^(+)CD86^(+),ESR,CRP,RF,Anti CCP Ab,IL-6,TNF in patients with rheumatoid arthritis-αThe levels of circ-CBLB,CD14^(+)CD163^(+),IL-4,and IL-10 were significantly higher than those of healthy individuals;(2)The expression level of circ-CBLB in patients with rheumatoid arthritis is positively correlated with CD14^(+)CD163^(+),and negatively correlated with miR-486-5p and CD14^(+)CD86^(+);The expression level of miR-486-5p is negatively correlated with CD14^(+)CD163^(+)and positively correlated with CD14^(+)CD86^(+);There is a negative correlation between CD14^(+)CD86^(+)and CD14^(+)CD163^(+);ESR is negatively correlated with circ-CBLB,and positively correlated with miR-486-5p,CD14^(+)CD86^(+),CRP;CRP is negatively correlated with circ-CBLB,CD14^(+)CD163^(+),and positively correlated with CD14^(+)CD86^(+),ESR;(3)The expression level of circ-CBLB in patients with rheumatoid arthritis is negatively correlated with joint tenderness,morning stiffness,lack of qi and lazy speech,and postprandial abdominal distension score;The expression level of miR-486-5p is positively correlated with the scores of joint tenderness and decreased appetite.(4)The ROC curve shows that in terms of circ-CBLB,ESR,CRP,VAS,and DAS28 AUC are 0.662(P=0.032),0.658(P=0.035),0.516(P=0.830),and 0.791(P=0.000),respectively.In terms of miR-486-5p,ESR,CRP,VAS,and DAS28 AUC were 0.566(P=0.385),0.511(P=0.883),0.592(P=0.223),and 0.727(P=0.003),respectively.Conclusion:The abnormal expression of circ CBLB and miR-486-5p in peripheral blood mononuclear cell of patients with rheumatoid arthritis of spleen deficiency and dampness excess type is related to inflammatory polarization markers,immune inflammation,disease activity,joint symptoms and spleen deficiency syndrome of rheumatoid arthritis,and the low expression of circ CBLB and high expression of miR-486-5p have certain predictive value for disease activity of rheumatoid arthritis.展开更多
OBJECTIVE:To investigate the effect of slow transit constipation(STC)and spleen Qi deficiency on gut microbiota,and the mechanism underlying the action that the positive drug Maren Runchang(MR)alleviates STC.METHODS:S...OBJECTIVE:To investigate the effect of slow transit constipation(STC)and spleen Qi deficiency on gut microbiota,and the mechanism underlying the action that the positive drug Maren Runchang(MR)alleviates STC.METHODS:STC was induced,using the cathartic method of Senna and the hunger-fullness disorder method,in ICR mice;one group of model mice was treated with MR(6.24 g/kg).The changes in the general condition,fecal parameters,D-xylose content in the serum,intestinal propulsion rate,and histopathology of the colon were assessed after STC induction in the control,model,and MR groups.Fecal microbiota transplantation(FMT)was performed from STC mice into pseudo germ-free mice.Changes in the contents of substance P(SP),vasoactive intestinal peptide(VIP),and gut microbiota in STC mice and pseudo germ-free mice were assessed after FMT.RESULTS:Compared with the control group,the model mice showed the following results:the time of the first black stool was significantly longer(P<0.01),the number and weight of black stools were significantly reduced within 6 h(P<0.05),the D-xylose content in the serum was significantly reduced(P<0.05),the intestinal propulsion rate decreased(P<0.01),the content of VIP in colon tissue significantly increased(P<0.05),and SP content in the colon tissue significantly decreased(P<0.01);moreover,the colon showed significant inflamemation and injury.Furthermore,the abundance of Firmicutes was increased,the abundance of Bacteroides decreased,and the abundance of Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus decreased,while the abundance of the conditional pathogenic bacteria Escherichia and Klebsiella increased.However,after treatment with MR,the time of the first black stool decreased(P<0.01),the number of black stools within 6 h increased,and the intestinal propulsion rate increased(P<0.05).Moreover,the content of D-xylose in the serum and the content of VIP in colon tissue significantly decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and colon inflammation significantly improved.Additionally,the abundance of Firmicutes decreased,and the abundance of Bacteroides increased.The abundance of unclassified_o_Bacteroidales and Bilophila increased,and the abundance of Klebsiella decreased.In the model+FMT group,compared with control+FMT group,the content of VIP in colon tissue decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and the abundance of probiotics,such as Lactobacillus and Akkermansia,decreased.In the MR+FMT group,compared with the model+FMT group,the content of VIP in colon tissue increased,the content of SP in colon tissue significantly decreased(P<0.01),and the abundance of probiotics increased.CONCLUSIONS:STC mice with spleen Qi deficiency show a decreased abundance of beneficial bacteria,such as Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus,and an increased abundance of the conditional pathogenic bacteria Escherichia coli and Klebsiella.Furthermore,the mechanism of action of MR in treating STC may involve the regulation of intestinal movement,reduction of intestinal inflammation,elevation of intestinal absorption,and regulation of gut microbiota.展开更多
基金the Seventh Batch of Inheritance Project of Academic Experience of National Veteran TCM Experts of the National Administration of Traditional Chinese Medicine (GZYYRJH[2022]No. 76)National Natural Science Foundation General Project (No.81973655)+4 种基金National Natural Science Foundation General Project (No.82274501)Anhui Provincial Excellent Youth Talent Support Program for Universities (Anhui Education Secretary [2022]No. 11)Key Laboratory of Xin'an Medical Education Department (No.2020xayx04)Natural Science Research Project of Universities in Anhui Province (No.KJ2020A0397)Anhui NewEra Education Quality Engineering Project (No.2022xscx103)。
文摘Objective:To explore the abnormal expression and significance of circ-CBLB/miR-486-5p in patients with rheumatoid arthritis of spleen deficiency and dampness excess type.Methods:The 30 healthy individuals included in the method were from the Physical Examination Center of Anhui Provincial Hospital of Traditional Chinese Medicine,and the 60 rheumatoid arthritis patients included were from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine.The disease activity score of patients with rheumatoid arthritis was evaluated using VAS score and DAS28 score,joint symptoms and spleen deficiency syndrome score were evaluated using graded quantification method,immune inflammation indicators were detected using relevant instruments,inflammatory cytokines were detected using ELISA method,macrophage markers were detected using FCM method,and pathway gene expression was detected using RT-qPCR;Evaluate the predictive effect of circ-CBLB and miR-486-5p on disease activity in rheumatoid arthritis using ROC curves.Results:(1)miR-486-5p,CD14^(+)CD86^(+),ESR,CRP,RF,Anti CCP Ab,IL-6,TNF in patients with rheumatoid arthritis-αThe levels of circ-CBLB,CD14^(+)CD163^(+),IL-4,and IL-10 were significantly higher than those of healthy individuals;(2)The expression level of circ-CBLB in patients with rheumatoid arthritis is positively correlated with CD14^(+)CD163^(+),and negatively correlated with miR-486-5p and CD14^(+)CD86^(+);The expression level of miR-486-5p is negatively correlated with CD14^(+)CD163^(+)and positively correlated with CD14^(+)CD86^(+);There is a negative correlation between CD14^(+)CD86^(+)and CD14^(+)CD163^(+);ESR is negatively correlated with circ-CBLB,and positively correlated with miR-486-5p,CD14^(+)CD86^(+),CRP;CRP is negatively correlated with circ-CBLB,CD14^(+)CD163^(+),and positively correlated with CD14^(+)CD86^(+),ESR;(3)The expression level of circ-CBLB in patients with rheumatoid arthritis is negatively correlated with joint tenderness,morning stiffness,lack of qi and lazy speech,and postprandial abdominal distension score;The expression level of miR-486-5p is positively correlated with the scores of joint tenderness and decreased appetite.(4)The ROC curve shows that in terms of circ-CBLB,ESR,CRP,VAS,and DAS28 AUC are 0.662(P=0.032),0.658(P=0.035),0.516(P=0.830),and 0.791(P=0.000),respectively.In terms of miR-486-5p,ESR,CRP,VAS,and DAS28 AUC were 0.566(P=0.385),0.511(P=0.883),0.592(P=0.223),and 0.727(P=0.003),respectively.Conclusion:The abnormal expression of circ CBLB and miR-486-5p in peripheral blood mononuclear cell of patients with rheumatoid arthritis of spleen deficiency and dampness excess type is related to inflammatory polarization markers,immune inflammation,disease activity,joint symptoms and spleen deficiency syndrome of rheumatoid arthritis,and the low expression of circ CBLB and high expression of miR-486-5p have certain predictive value for disease activity of rheumatoid arthritis.
基金Supported by Fundamental Scientific Research Business Fees of Central Public Welfare Scientific Research Institutes of China Academy of Chinese Medical Sciences(No.Z2021022)National Natural Science(No.81891010):Study on the Authenticity of Traditional Chinese Medicine,Science and Technology Innovation Project of China Academy of Chinese Medical Sciences:Efficacy Evaluation and Mechanism Exploration of Huashi Baidu Granule based on"multi-link and multi-target"in the Prevention and Treatment of Influenza Virus Infection(CI2021A04605)Basic Scientific Research Business Fee Project of Central Public Welfare Scientific Research Institutes of the Chinese Academy of Chinese Medical Sciences:Study on the Novel Coronavirus Pneumonia Mechanism of Yiqi Zeng Fang and Huashi Baidu Granules(ZZ15-WT-04)
文摘OBJECTIVE:To investigate the effect of slow transit constipation(STC)and spleen Qi deficiency on gut microbiota,and the mechanism underlying the action that the positive drug Maren Runchang(MR)alleviates STC.METHODS:STC was induced,using the cathartic method of Senna and the hunger-fullness disorder method,in ICR mice;one group of model mice was treated with MR(6.24 g/kg).The changes in the general condition,fecal parameters,D-xylose content in the serum,intestinal propulsion rate,and histopathology of the colon were assessed after STC induction in the control,model,and MR groups.Fecal microbiota transplantation(FMT)was performed from STC mice into pseudo germ-free mice.Changes in the contents of substance P(SP),vasoactive intestinal peptide(VIP),and gut microbiota in STC mice and pseudo germ-free mice were assessed after FMT.RESULTS:Compared with the control group,the model mice showed the following results:the time of the first black stool was significantly longer(P<0.01),the number and weight of black stools were significantly reduced within 6 h(P<0.05),the D-xylose content in the serum was significantly reduced(P<0.05),the intestinal propulsion rate decreased(P<0.01),the content of VIP in colon tissue significantly increased(P<0.05),and SP content in the colon tissue significantly decreased(P<0.01);moreover,the colon showed significant inflamemation and injury.Furthermore,the abundance of Firmicutes was increased,the abundance of Bacteroides decreased,and the abundance of Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus decreased,while the abundance of the conditional pathogenic bacteria Escherichia and Klebsiella increased.However,after treatment with MR,the time of the first black stool decreased(P<0.01),the number of black stools within 6 h increased,and the intestinal propulsion rate increased(P<0.05).Moreover,the content of D-xylose in the serum and the content of VIP in colon tissue significantly decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and colon inflammation significantly improved.Additionally,the abundance of Firmicutes decreased,and the abundance of Bacteroides increased.The abundance of unclassified_o_Bacteroidales and Bilophila increased,and the abundance of Klebsiella decreased.In the model+FMT group,compared with control+FMT group,the content of VIP in colon tissue decreased(P<0.05),the content of SP in colon tissue significantly increased(P<0.01),and the abundance of probiotics,such as Lactobacillus and Akkermansia,decreased.In the MR+FMT group,compared with the model+FMT group,the content of VIP in colon tissue increased,the content of SP in colon tissue significantly decreased(P<0.01),and the abundance of probiotics increased.CONCLUSIONS:STC mice with spleen Qi deficiency show a decreased abundance of beneficial bacteria,such as Alistipes,unclassified_o_Bacteroidales,Alloprevotella,Bilophila,and Anaerotruncus,and an increased abundance of the conditional pathogenic bacteria Escherichia coli and Klebsiella.Furthermore,the mechanism of action of MR in treating STC may involve the regulation of intestinal movement,reduction of intestinal inflammation,elevation of intestinal absorption,and regulation of gut microbiota.
