| ObjectiveObserve the distribution of different types of RA and its relationship with small plate activation,summarize the TCM syndrome characteristics of RA platelet activation,and explore the distribution of TCM syndromes of rheumatoid arthritis(RA)platelet activation.According to the characteristics of common TCM syndromes of RA platelet activation,the use of TCM intervention is used to verify the TCM syndrome characteristics of RA platelet activation,in order to guide clinical syndrome differentiation and treatment,so as to more effectively choose the TCM treatment plan for prevention and treatment.MethodsRandomly select 100 patients with RA in outpatient and inpatient clinics of the First and Third Affiliated Hospitals of Anhui University of Traditional Chinese Medicine.Based on the actual clinical syndrome types,the TCM syndrome differentiation is based on the clinical symptoms and signs.Four common syndrome types are summarized: liver and kidney yin deficiency syndrome,damp-heat arthralgia syndrome,cold-dampness arthralgia syndrome,phlegm stasis arthralgia syndrome.According to the diagnostic criteria,a unified table was designed for observation.The research content included the patient’s gender,age,medical history,symptoms,signs,tongue and pulse.At the same time,observe the changes of RA patients’ joints,systemic symptoms scores,quality of life scores,health assessment index(HAQ),disease activity score(DAS-28),pain score(SAS),laboratory indicators and platelet activation.According to the characteristics of RA clinical TCM syndromes,clinical medications for the treatment of RA platelet activation are drawn.Animal experiments have verified the characteristics of TCM syndromes of RA platelet activation.The rats were randomly divided into 4 groups: normal control(NC)group,model control(MC)group,leflunomide(LEF)group,and Xinfeng capsule(XFC)group,each with 10 rats.Except for the NC group,each animal was injected intracutaneously with 0.1ml of Freund’s complete adjuvant(FCA)into the right hindfoot plantar to induce inflammation,which was replicated as an AA model.The drug was administered on the12 th day after inflammation,and the drug dose of each group was equivalent to 10 times the clinical dose.The doses of each group are as follows: NC group: normal saline gavage,1ml/100 g,once a day;MC group: normal saline gavage,1ml/100 g,once a day;LEF group: LEF plus normal saline for preparation Into a suspension(with LEF0.3mg per ml),gavage 1ml/100 g once a day;XFC group: remove XFC from the capsule shell,add physiological saline,and prepare a suspension(each ml contains 0.3g drug),according to the dose of 1ml/100 g,once a day.All groups were given intragastric administration for 4 weeks.Observe the swelling degree and arthritis index of each group of rats.ELISA method is used to determine the expression of IL-10 and IL-21 in rat serum.The expression of GMP-140 and CD40 L in peripheral blood is detected by flow cytometry.Reverse transcription polymerase chain method The reaction was used to detect the expression of ETS-1 and PAF mRNA in rat synovial tissue.Western blotting was used to detect the expression of ETS-1 and PAF protein in rat synovial tissue.ResultThe clinical research results show:(1)Changes in the frequency(rate)of abnormal platelet parameters of each group of syndrome types: The most abnormal number of platelet parameters is the syndrome of damp-heat arthralgia syndrome.Among them,the abnormal rate of PLT decreased to damp-heat arthralgia syndrome,cold-dampness arthralgia syndrome,phlegm stasis syndrome,liver and kidney deficiency syndrome;the MPV abnormal rate decreased to liver and kidney deficiency syndrome,damp-heat arthralgia syndrome,phlegm stasis Arthralgia syndrome,cold-damp arthralgia syndrome;PCT abnormal rate decreased to damp-heat arthralgia syndrome,liver and kidney deficiency syndrome,phlegm-stasis arthralgia syndrome,cold-dampness arthralgia syndrome;PDW abnormal rate decreased successively to phlegm-stasis arthralgia syndrome,Damp-heat arthralgia syndrome,liver and kidney deficiency syndrome,cold-dampness arthralgia syndrome.(2)Comparison of platelet parameters: PLT of damp-heat arthralgia syndrome is higher than the other three syndromes(P<0.01 or P<0.05),PLT of cold-dampness arthralgia syndrome,phlegm stasis arthralgia syndrome is higher than liver and kidney deficiency syndrome High(P<0.01).The PDW of cold-damp arthralgia syndrome was higher than that of liver and kidney deficiency syndrome(P<0.05).(3)Comparison of quality of life scores: Compared with liver and kidney deficiency syndrome,damp-heat arthralgia syndrome,cold-dampness arthralgia syndrome,phlegm-stasis arthralgia syndrome,physiological function scores increased,and damp-heat arthralgia syndrome syndrome increased self-awareness(P< 0.01 or P<0.05).Compared with damp-heat arthralgia syndrome,the physiological function score of phlegm-stasis arthralgia syndrome decreased(P<0.05).(4)Comparison of joint and systemic symptom scores: Compared with liver and kidney deficiency syndrome,damp-heat arthralgia syndrome,cold-dampness arthralgia syndrome,joint pain,joint tenderness,loss of appetite,increased abdominal distension score after eating(P<0.05),sputum The morning stiffness score of blood stasis syndrome increased(P < 0.05).Compared with damp-heat arthralgia syndrome and cold-damp arthralgia syndrome,the joint pain score of phlegm-stasis arthralgia syndrome decreased(P<0.05).(5)Comparison of HAQ,DAS-28,and VAS: Compared with liver and kidney deficiency syndrome,HAQ,DAS-28,and VAS scores increased(P<0.01 or P<0.05).The VAS score of the syndrome of phlegm stasis and arthralgia increased(P<0.05).Compared with the syndrome of damp-heat arthralgia syndrome,the HAQ score of the syndrome of phlegm-stasis arthralgia increased,and the DAS-28 score decreased(P<0.01 or P<0.05).Compared with the syndrome of cold-dampness blockage,DAS-28 score of phlegm-stasis blockage syndrome decreased(P<0.05).(6)Comparison of laboratory indicators: Compared with liver and kidney deficiency syndrome,RF,CCP,CRP,ESR of damp-heat arthralgia syndrome increased(P<0.01 or P<0.05),and RF,CRP,ESR of cold-damp arthralgia syndrome increased.High(P<0.01),RF and ESR of phlegm-stasis-bi-block syndrome increased(P<0.05).Compared with damp-heat arthralgia syndrome,CCP of cold-damp arthralgia syndrome decreased(P<0.05),and RF,CCP,CRP,ESR of phlegm-stasis arthralgia syndrome decreased(P<0.01).Compared with the syndrome of cold-dampness blockage,the RF,CRP and ESR of the syndrome of phlegm-stasis blockage decreased(P<0.01 or P<0.05).(7)Comparison of platelet activator GMP-140,PAF and ETS-1: And in comparison between groups,GMP-140 and PAF of damp-heat arthralgia syndrome were higher than the other three syndromes(P<0.01 or P<0.05).The PAF of the syndrome of cold-dampness blockage,phlegm-stasis blockage syndrome was higher than that of liver and kidney deficiency syndrome(P<0.01 or P<0.05).The ETS-1 of the syndrome of phlegm stasis and arthralgia was lower than that of the deficiency of liver and kidney(P<0.05).(8)Correlation analysis between platelet activators and other indicators of different types of RA: liver and kidney deficiency syndrome GMP-140 is positively correlated with healthy self-recognition and Ig G,PAF is positively correlated with joint morning stiffness,ETS-1 is positive with CCP-AB Correlation(P<0.05).GMP-140 was negatively correlated with PCT(P < 0.05).GMP-140 was positively correlated with joint pain,abdominal distension after eating,PLT,and CRP,and ETS-1 was negatively correlated with loose stools,DAS-28 and ESR(P<0.01 or P<0.05).PAF was negatively correlated with physiological function,joint pain,fatigue,PCT,and RF(P<0.01 or P<0.05).Cold-dampness syndrome GMP-140 was positively correlated with joint swelling and loose stools,PAF was positively correlated with loss of appetite,MPV,and ETS-1 was positively correlated with joint morning stiffness(P<0.05).Phlegm-stasis-blocking syndrome GMP-140 was positively correlated with loose stools and PLT,PAF was positively correlated with joint morning stiffness,CCP-AB,and ETS-1 was positively correlated with joint swelling(P<0.05).Animal research shows:(1)Observation of arthritis model and results of drug intervention:After modeling,rats with adjuvant arthritis developed red and swollen limbs,joints,ulceration of metatarsophalangeal toes,dry and sparse fur,curled up and less movement,squinted eyes,sluggishness,listlessness,slow response,various degrees of swelling,and decreased body mass.The amount of food and drinking water decreased,and some rats had loose stools.The administration started on the 12 th day.On the 25 th day,the redness and swelling of the limbs and joints of the rats in the treatment group gradually subsided,their body mass,diet and water intake began to increase,and their spirits improved and became active.On the 35 th day,rats in each treatment group had shiny hair and active behavior.On the 40 th day,the joint and systemic symptoms of the rats in each treatment group basically disappeared,and the body weight,diet and water intake still increased.Among them,the XFC group was more significant than the MTX group.The state of rats in the normal control group did not change significantly.(2)The effect of Xinfeng Capsule on the swelling degree and arthritis index of the sole:Before administration,compared with the NC group,the plantar swelling and arthritis index of rats in the MC group,LEF group and XFC group were significantly increased(P<0.01).After administration,compared with the MC group,the plantar swelling and arthritis index of the LEF group and XFC group were significantly reduced(P<0.01 or P<0.05).(3)The effect of Xinfeng Capsule on serum IL-10,IL-21,ETS-1,PAF:Compared with the NC group,the cytokines IL-21 and PAF were significantly increased in the MC group,and the cytokines IL-10 and ETS-1 were significantly decreased(P<0.05 or P<0.01).Compared with the MC group,the expression of cytokines IL-21 and PAF in the XFC group decreased,and the expression of cytokines IL-10 and ETS-1 increased(P <0.05 or P <0.01).(4)The effect of Xinfeng Capsule on peripheral blood GMP-140 and CD40L:Compared with the NC group,the peripheral blood GMP-140 and CD40 L of the MC group increased significantly(P<0.01).Compared with the MC group,the peripheral blood GMP-140 and CD40 L in the LEF group and XFC group were significantly reduced(P<0.01 or P<0.05).(5)The effect of Xinfeng Capsule on synovial ETS-1 and PAF mRNA: Compared with NC group,the expression of ETS-1 mRNA in MC group decreased,and the expression of platelet activation product PAF mRNA and increased.Compared with the MC group,ETS-1mRNA increased in LEF and XFC groups,while PAFmRNA decreased(P<0.01 or P<0.05).(6)The relationship between platelet activator and plantar swelling degree,arthritis index,IL-10,IL-21,ETS-1 mRNA/protein: GMP-140 is positively correlated with plantar swelling degree,CD40 L and IL-21 are positive Correlation,PAF mRNA was positively correlated with plantar swelling(P<0.01 or P<0.05).ConclusionThe TCM syndromes of RA are mainly damp-heat arthralgia,accompanied by phlegm stasis arthralgia,and platelet activation is involved in the process of RA damp-heat arthralgia,cold dampness arthralgia,and phlegm stasis arthralgia,and spleen deficiency and dampness prevails across RA.By regulating the expression of ETS-1,Xinfeng Capsule,a traditional Chinese medicine for invigorating spleen,dampness and dredging,reduces platelet activation,inhibits immune inflammatory response,reduces immune complex deposition in synovial tissue,and improves RA joint symptoms. |
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