| Objective: To evaluate the clinical efficacy and explore the impact on immune homeostasis of replenishing qi,expelling dampness and dissipating blood stasis comprehensive therapy of traditional Chinese medicine(REDCT)to Toll like receptors(TLRs)—Myeloid differentiation factor 88(MyD88)/ Growth arrest-specific gene 6(Gas6)— Tyro3/Axl/Mer Receptor signaling pathways in patients with recurrent pelvic inflammatory disease(RPID).Methods:1.Prospective observational studies,self-controlled trial was designed.108 patients from Hospital of Chengdu university of TCM who matched the inclusion criteria were included.Replenishing qi,expelling dampness and dissipating blood stasis comprehensive therapy of traditional Chinese medicine(REDCT)was used totally for 3 months.The pelvic pain symptom score,pelvic physical examination score,TCM syndrome score and PID recurrence rate were used to evaluate the clinical efficacy at the beginning,after 4 weeks’,8 weeks’ and 12weeks’ treatment,12 weeks follow up.2.Immunological indicators were tested in 100 RPID patients who completed the treatment regimen and 50 healthy subjects(HS).Flow cytometry,RT-PCR,ELISA laboratory tests were used to detect the TLR2,TLR4 mRNA and protein expression in peripheral blood,and MyD88,Gas6,sMer,sAxl,IL-17,IL-23 in serum of HS and RPID patients before and after the treatment(at 12 weeks).Binary logistic regression analysis and Receiver operating characteristic curve(ROC)were also used to screen out the factors that may affect the incidence of RPID immune disorders.Results:1.The clinical efficacy of REDCT: The REDCT could significantly reduce the pelvic pain symptom score,TCM syndrome score and pelvic physical examination score of RPID patients,which were significantly different from the baseline before treatment(P<0.001).Analysis based FAS at the end of treatment(12 weeks after treatment): the rate of pelvic pain relief was 77.78%.The recovery and significant efficiency rate of pelvic physical examination were 61.11%(66/108 cases)and the effective rate was 97.22%(105/108 cases).The recovery and significant efficiency rate of TCM syndrome were 73.14%(79/108 cases),the effective rate was 96.30%(104/108 cases).In 3 months of follow-up,the recurrence rate was 3.79%.The results of PPS and FAS are consistent.2.The impact on immune homeostasis of REDCT1)TLR2 and TLR4 were both expressed on Monocytes,lymphocytes and granulocytes of RPID patients’ peripheral blood.The relative expression levels of TLR2,TLR4 protein and mRNA in monocytes were significantly different from that in HS(P<0.05),and were positively correlated with the disease condition(P<0.05).The levels of MyD88,Gas6,sMer,sAx1,IL-17,and IL-23 in the serum of RPID patients were all abnormally expressed.The levels of MyD88,IL-17,and IL-23 were significantly increased compared with HS(P<0.05).The expression levels of IL-17 and MyD88 were positively correlated with the disease condition and course of disease,respectively(P<0.05).Compared with the HS,the levels of Gas6,sMer and sAxl in RPID patients were all decreased.Except for sAxl,the differences of Gas6 and sMer were statistically significant(P<0.05),and the levels of Gas6 showed a significant negative correlation with the disease condition and course of disease(P<0.05).Logistic regression analysis showed that the increased expressions of TLR2 mRNA,TLR4 mRNA,MyD88 and IL-17 may lead to a corresponding multiple increase in the risk of RPID,and the corresponding OR values were 1.586,1.649,1.550 and 1.094,respectively(P<0.05).Decreased expressions of Gas6 and sMer may lead to a corresponding multiple increase in the risk of RPID,and the corresponding OR values were 0.749 and 0.973(P<0.05).2)The ROC curve analyzed the potential diagnostic predictive efficacy of the above-mentioned items.The results showed that the area under the curve of Gas6,IL-17 and combined detection were all >0.7,but <0.9,each index had certain clinical diagnostic efficacy,and the sensitivity of IL-17 was 0.68,the specificity was 0.78;the sensitivity of Gas6 was 0.68,the specificity was 0.76;the sensitivity of Gas6/IL-17 combined detection was 0.80,the specificity was 0.89.Among them,the combined detection has the highest AUC and high specificity.3)After treatment,the levels of the above indicators were improved and were close to HS: the levels of TLR2 and TLR4 mRNA and protein expression were significantly down-regulated than baseline(P<0.05);MyD88,IL-17,IL-23 levels were significantly down-regulated than baseline(P<0.05);There were positive correlation between the differences in the levels of Myd88,IL-17 and the improvement of disease(P<0.05).Gas6,sMer levels were significantly up-regulated than baseline(P<0.05).Conclusions:1.The clinical efficacy of REDCT is better than similar trials,especially in relieving the symptoms of repeated lower abdominal pain and promotingthe recoveryof TCM syndromeand the pelvic physical signs,and a higher rate of recurrence control.2.The imbalance of immune homeostasis which was caused by TLR2,TLR4,MyD88,Gas6,sMer,sAx1,IL-17,IL-23 may lead to RPID.3.REDCT may downgrade the expression of TLR2,TLR4,MyD88,IL-17 and IL-23 and promote the expression of Gas6,sMer in peripheral blood.TLR2,TLR4,Gas6,MyD88,and IL-17 may be molecular targets for REDCT to effectively improve the prognosis of RPID.4.The combined detection of three indicators(Gas6/IL-17)has a certain potential predictive effect on the onset of RPID and has clinical diagnostic value.This study provides a new basis for the establishment of RPID scientific objective evaluation system. |
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