The Mechanism Of Banxia Qinlian Decoction In Inhibiting The Immune Inflammatory Injury Of Exocrine Glands In Sjogren's Syndrome And Its Clinical Efficacy Analysis

Sjogren's Syndrome(SS)is a chronic autoimmune disease.It mainly invades the exocrine glands,especially salivary and lachrymal glands,causing dryness in mouth,eyes,and skin.The clinical manifestations of SS can involve musculoskeletal system,respiratory system,digestive system,urinary system,cardiovascular system,hematological system and nervous system.The exact pathogenesis of this disease is still unknown.Also it lacks of effective and safe therapy.Traditonal Chinese Medecine(TCM)has the advantages in regulating the body's immune function and Relieving clinical symptoms.According to years of clinical experience,Profesor Meng Fengxian considered the disorder of Qi's movement as an important role in the pathogenesis of SS.Based on this theory,she created Banxiaqinlian Decoction(BXQLD)to treat SS patients,which getting good curatice effects in clinic.This research discussed the pathogenesis of SS and the curative effect and immunological inflammatory mechanism of BXQLD for treatment of SS by three parts as follow.Part 1 Literature reviewThis part reviewed the recent advance in the pathogenesis,pathology,clinical manifestations,diagnoses and therapies management of SS from the both aspects of traditional Chinese medicine and modern medicine.It also review the research advances in animal models of human SS.Part 2 animal experimentObject To investigate the the immuno-inflammatory mechanism of exocrine gland injury and the mechanism of water transport.To explore the effects of BXQLD on the mechanism above.Materials and methods 24 female NOD mice were randomly divided into 4 groups,i.e.,the model group,the positive drug group and the high-,medium-dose BXQLD groups,6 in each group.Another 6 female Balb/c mice were taken as the noma1 group.Glucosidorum Tripterygll Totorum(TII)was intragastrically administered at the daily dose of 10mg/kg to mice in the positive drug group.BXQLD was intragastrically administered at the daily dose of 120g/kg,60g/kg to mice in the high-,medium-dose BXQLD groups.Distilled water was intragastrically administered to mice in the normal control group and the model group everyday.The experiment lasted for 12 weeks.During the experiment,the weight and food intake of each mouse were observed every 4 weeks.Mice of each group were executed at the end of 12th week.Their parotid glands and submandibular glands were taken out to determine the organ index and morphology of salivary glands.The mRNA transcription level of AMPK?PPAR-??M3R?AQP5?RANTES?MiP-1??IL-17?IL-6?TNF-???-fodrin in salivary glands tissue were determined by reverse transcription-polymerase chain reaction(RT-PCR).The protein expression levels of AMPK?1?M3R?AQP5 in salivary glands tissue were determined by Western blot.We also determined the serum content of SSA?SSB?ANA?MiP-1??TNF-??IL-17?IL-6.Results There were totally 6 NOD mice dead at the end of experiment,including 2 in the model group,1 in the positive drug group and 4 in high-dose BXQLD group.There was no.statistical difference in the weight and food intake among all groups during the experiment.At 12th week:(DThere was no statistical difference in the organ index among all groups during the experiment(F>0.05)�?Compared with normal group,NOD mice in model group was obviously infiltrated by lymphocyte(P<0.05).BXQLD showed increasing tendency to decrease lymphocytic infiltration into salivary glands.?The mRNA transcription level of AMPK of salivary gland tissue in model group were lower than normal group(P<0.01),while PPAR-??M3R?AQP5?RANTES?IL-17?IL-6?TNF-??MIP-1? were much higher(P<0.01).There was no statistical difference in the mRNA transcription of a-fodrin among all groups(P>0.05).Compared with model group,BXQLD showed up-regulated transcription of AMPK mRNA and down-regulated transcription of PPAR-?,M3R and MIP-1?(P<0.05,P<0.01).The high-dose BXQLD group showed down-regulated expression of AQP5 mRNA(P<0.05),while medium-dose BXQLD group showed no significant diference compared with model group(P>0.05).? Compared with normal group,the protein expression of AMPK?1 significantly decreased(P<0.05),but both M3R and AQP protein expression increased obviously(P<0.01).BXQLD could increase the protein expression of AMPK?1(P<0.05).High-dose BXQLD group showed down-regulated expression of AQP5 protein(P<0.05),while medium-dose BXQLD group showed no significant diference compared with model group(P>0.05)..?Compared with normal group,the serum content of SSA?SSB?TNF-??IL-6?IL-17 increasedin the model group(P<0.05,P<0.01),while the serum content of ANA and Mip-1? ahad no significant swings(F>0.05).BXQLD could reduce the expression of SSA?SSB?TNF-??IL-6?IL-17 in serum(P<0.05,P<0.01).Conclusions?BXQLD could protect salivary glands from immuno-inflammatary injury by not only inhibiting the expression of pro-inflammatory factors(IL-6?IL-17?TNF-?)and chemotactic factor(RANTES),but also stimulating the expression of anti-inflammatory factor AMPK.?The up-regulated expression of M3R and AQP5 was related with improving the ability of water transport,or participating in the mechanism of inflammatory injury.High-dose BXQLD could decrease the expression of M3R and AQP5.Part 3 Clinica1 research 1Object To study Chinese medical pattern characteristic of SS and to explore the pathogenesis of SSMethods 108 outpatients of SS from Octorber 2011 to March 2014 in department of rheumatology in Dongfang Hospital were enrolled in the research.We colleted the data of patients' basic information,current therapy,clinical symptoms,the tongue picture and pulse at their fisrt visit.Then according to the clinical symptoms,we analysed the patterns of syndrome,and discussed pathogenesis of 5S.Result ? Patients with basic information was as follow.Among 108 patients,there were 104 females and 4 males.The average age was 54.37.There were 16 patients combined with other rheumatic diseases and 18 patients treated by anti rheumatic drugs in werstern medicine.? Clinical features was as follow.Among 108 cases,symptoms of dry mouth and dry eyes appeared most frequently,while the symptoms of pain joint,fatigue,poor appetite,stomach bloating and sore joint followed.? Tongue picture was as follow.Red tongue appears highest rate of 78 cases(72.22%),followed by pink tongue,dark tongue,pale tongue.The thin greasy tongue coating appeared the highest rate of 60 cases(55.56%),followed by the tongue with thick and greasy coating,less or no coating,thin dry coating and normal coating.Except 10 patients with little or no tongue coating,the color of 83 patients' tongue coating was white,while other 15 yellow.?Pulse distribution was as follow.Slide pulse appeared most frequently in 108 patients.? Pattern of TCM syndromes was as follow.57 cases belonged in Yin-deficiency with phlegm dampness,while 20 in Syndrome of Qi and damp stagnation,13 in phlegm-accumulation stasis,10 in Qi and Yin defeciency.Deficiency of yin and excessive heat syndrome,Yin-deficiency and blood stasis,single Yin-deficiency syndrome appeared in 4-,3-,1-,cases,respectively.?Distribution of syndrome factors was as follow.Yin-deficiency appeared in 100 cases.Phlegm and dampness appeared in 95 cases.Qi-stagnation appeared in 85 cases.Qi-deficiency and blood-stasis both appeared in 32 cases.Heattoxicity appeared in 7 cases cold.Except 1 cases with single factors,all patients appeared more than 2 syndrome factors at the same time.Conclusion Disorder of qi and phlegm-dampness may be one of the pathogenesis of SS.Part 4 Clinical research 2Object To observe the curative effect of BXQLD and to explore the mechanism of BXQLD.Methods 45 SS outpatients,who treated by BXQLD for at less 3 weeks from Octorber 2011 to March 2014 in department of rheumatology in Dongfang Hospital,were enrolled in the research.We colleted the data of patients' basic information,current therapy,clinical symptoms,the tongue picture and pulse at their f isrt and last visit.Then we conducted the clinical symptoms of pat ients before and after treatment to TCM syndrome score.According to the overall score,dry mouth score,dry eyes scores and other secondary symptoms scores,we observed the effect of BXQLD and the relationship between the curative effect and both course of treatment and tongue characteristics.Result ? Patients with basic information was as follow.45 patients were all females,with an average age of 53.58,The average course of treatment was 32.4 weeks.? Clinical features was as follow.43 in 45 cases appeared symptoms of both dry mouth and dry eyes,with the highest rate of appearance.The rate of dry nose,dry skin,dry stool proportion appearance was all obove 20%.In addition to symptoms of dryness,stomach bloating,pain joint,poor appetite,fatigue,sore joint were also the common symptoms.?Tongue picture was as follow.Red tongue appeared highest rate of 29 cases(64.44%),followed by pink tongue,pale tongue,dark tongue.The thin greasy tongue coating appeared the highest rate of 26 cases(45.9%),followed by the tongue with thick and greasy coating,thin dry coating,less or no coating.Except 3 patients with little or no coat ing,the color of 32 pat ients tongue coating was white,while other 9 yellow.? Pulse distribution was as follow.Slide pulse appeared in 40 cases,which appeared most frequently in 45 patients.? Curative effect of BXQLD was as follow.After treatment,the score of entire symptoms,dry mouth,dry eyes and other main symptoms,pain joint,swelling and sore joint,poor appetite,fatigue,chest tightness were statistica11y decreased(P<0.05,P<0.01).There was no significant difference of the score of dry skin,dry stools,dry cough symptom(P>0.05).The effect of BXQLD was related with the course of treatment(P<0.01).There was no different effect to treat patients with different tongue picture by BXQLD.(P>0.05).Conclusion BXQLD could obviously relieve symptoms of SS patients.