Mechanism And Clinical Study Of Huayu Tongluo Method On Blood Stasis Microenvironment Of Pelvic Inflammatory Disease Sequelae

Objective:Sequelae of pelvic inflammatory disease is a common and frequently-occurring disease in clinical.Traditional Chinese medicine (TCM) has certain characteristic and advantage in treatment of this disease. TCM believes that blood stasisis the basicpathogenesis for the occurrence and development of this disease. On this basis, we propose the concept of blood stasismicroenvironment of sequelae of pelvic inflammatory disease. From this perspective, we formulated Danzhi Decoction to prescribe that stasis-reducing and venation-flowing is the basic way of TCM for treating this disease and it achieves good clinical curative effects. Methods:Absorption of oral TCM was simulated in cell experiment. Through 3D cell culture, co-culture system of human vascular endothelial cells/human endometrial stromal cells/humanendometrial glandular epithelial cells was established. And 3 groups including Danzhi Decoction group, aspirin group and the blank group were set. With Milliplex multifunctional liquid chip, the influences of Danzhi Decoction on 17 angiogenesis factors were primarily discussed and compared with aspirin group. Immunofluorescence combined with laser confocal microscopy and 3D reconstruction was used to find the possible site of Danzhi Decoction in improvement of sequelae of pelvic inflammatory disease. Patients with sequelae of pelvic inflammatory disease from Dongfang Hospital were selected as the research objects of clinical trials. With Danzhi Decoction as the basic prescription, which was adjusted according to the clinical syndrome, the patients were compared and analyzed in pretreatment and posttreatment pain scores, symptoms scores and TCM syndrome scores, in order to clarify the core role of stasis-reducing and venation-flowing method in treatment of sequelae of pelvic inflammatory disease. Results:Milliplex multifunctional liquid chip was used to compare vascular growth factors of each group after intervention. With regard to comparison between Danzhi Decoction and aspirin group, P>0.05 showed there were no statistically significant differences between Danzhi Decoction and aspirin group in vascular growth factors; After comparison between TCM group and the blank group, the most obviously changed vascular endothelial factors were VEGF-A, VEGF-C and VEGF-D; Through immunofluorescence combined with laser confocal microscopy and 3D reconstruction, positions with changes of VEGF-A, VEGF-C and VEGF-D were determined. The results showed that VEGF-A, VEGF-C and VEGF-D were most obviously changed in humanendometrial glandular epithelial cells; With regard to clinical findings, pain scores, symptoms scores and TCM syndrome scores were all significantly improved after treatment. In particular, pretreatment and posttreatment pain scores were 5.467±2.191 vs.2.667±1.859 (P<0.001); For symptoms scores, they were 9.422±3.577 vs.4.956±3.075 (P<0.001); while for TCM syndrome scores, they were 18.2±3.12 vs.6.6±1.92 (P<0.05) in cold and dampness stasis syndrome,17.93±3.08 vs.6.79±1.58 (P<0.05) in Qi stagnation and blood stasis syndrome.Conclusion:Both Danzhi Decoction with stasis-reducing and venation-flowing effects and western medicine aspirin can well reduce the angiogenesis factor in endometrial cells with blood stasismicroenvironment of sequelae of pelvic inflammatory disease. They have the equal effects on improvement of local blood stasismicroenvironment; Among the 17 angiogenesis factors, Danzhi Decoction has especially obvious effects on VEGF-A, VEGF-C and VEGF-D; the action targets may be located on humanendometrial glandular epithelial cells; Stasis-reducing and venation-flowing method is a basic thinking for treatment of sequelae of pelvic inflammatory disease, which is adjusted according to different syndrome types (cold and dampness stasis type, Qi stagnation and blood stasis type). And it has obvious improvement on clinical symptoms, signs and TCM syndromes.