Quercetin of huoxuehuayu tongluo decoction and azithromycin combination therapy effectively improves rat tubal factor infertility by inhibiting inflammation

Document Type : Original Article

Authors

1 Department of Emergency, the First Affiliated Hospital of Hunan Traditional Chinese Medicine College, Zhuzhou, Hunan, China

2 Department of TCM Gynaecology, the First Affiliated Hospital of Hunan Traditional Chinese Medicine College, Zhuzhou, Hunan, China

3 Combine traditional Chinese and Western Medicine Institute, Hunan University of Chinese Medicine, Changsha, Hunan, China

4 Clinical Laboratory, the First Affiliated Hospital of Hunan Traditional Chinese Medicine College, Zhuzhou, Hunan, China

Abstract

Objective(s): Tubal factor infertility (TFI) is common female infertility responsible for a large portion of female factor infertility. This study reveals the effect of the quercetin of Huoxuehuayu Tongluo Decoction with azithromycin on the pregnancy rate and inflammation of TFI female rats.
Materials and Methods: Female Sprague Dawley rats were constructed into the TFI model and treated with quercetin, Huoxuehuayu Tongluo Decoction, and combination therapy (quercetin and azithromycin). Pregnancy rate and litter size were measured. Network pharmacology was applied to analyze the interaction between Huoxuehuayu Tongluo Decoction and TFI. The combination of quercetin and IL-6 was analyzed by molecular docking. HE staining and electron microscopy were used to observe the histopathology and ultrastructure of fallopian tube tissues. The TNF-α, IL-1β, IL-6, IL-8, and MPO levels were detected by ELISA. The activation of JAK/STAT, MAPK, and NF-κB p65 pathways was detected by western blot or immunohistochemistry.
Results: Quercetin was the main active component of Huoxuehuayu Tongluo Decoction, and could bind to IL-6 in TFI. Target genes were enriched in the IL-17 signaling pathway, JAK-STAT signaling pathway, inflammatory disease, etc. Under the quercetin and azithromycin combination therapy, both rat pregnancy rates and litter sizes increased significantly. quercetin and azithromycin alleviated the symptoms of hydrosalpinx and inflammatory damage in fallopian tube tissues. The phosphorylation of JAK/STAT and MAPK pathways and NF-κB p65 translocation to the nucleus were significantly inhibited by the quercetin and azithromycin therapy.
Conclusion: Quercetin and azithromycin combination therapy inhibited inflammation and phosphorylation of JAK/STAT and MAPK pathways to improve TFI inflammation and pregnancy function.

Keywords

Main Subjects


1. Liao W, Tang X, Li X, Li T. Therapeutic effect of human umbilical cord mesenchymal stem cells on tubal factor infertility using a chronic salpingitis murine model. Arch Gynecol Obstet 2019; 300:421-429.
2. Tsevat DG, Wiesenfeld HC, Parks C, Peipert JF. Sexually transmitted diseases and infertility. Am J Obstet Gynecol 2017; 216:1-9.
3. Refaat B, Dalton E, Ledger WL. Ectopic pregnancy secondary to in vitro fertilisation-embryo transfer: Pathogenic mechanisms and management strategies. Reprod Biol Endocrinol 2015; 13:30-47.
4. Mardh PA. Tubal factor infertility, with special regard to chlamydial salpingitis. Curr Opin Infect Dis 2004; 17:49-52.
5. Savaris RF, Fuhrich DG, Duarte RV, Franik S, Ross J. Antibiotic therapy for pelvic inflammatory disease. Cochrane Database Syst Rev 2017; 4:CD010285.
6. Tsai WH, Yang CC, Li PC, Chen WC, Chien CT. Therapeutic potential of traditional chinese medicine on inflammatory diseases. J Tradit Complement Med 2013; 3:142-151.
7. Azawi OI, Al-Abidy HF, Ali AJ. Pathological and bacteriological studies of hydrosalpinx in buffaloes. Reprod Domest Anim 2010; 45:416-420.
8. Price MJ, Ades AE, Welton NJ, Simms I, Macleod J, Horner PJ. Proportion of pelvic inflammatory disease cases caused by chlamydia trachomatis: Consistent picture from different methods. J Infect Dis 2016; 214:617-624.
9.Yin Y, Shao L, Zhang L, Lei L, Liang C, Liu L. Effects of fallopian tube recuperation soup on JAK 2 / STAT 3 pathways in rats with tubal infertility. Int J Clin Exp Med 2018; 11:10856-10864.
10. Zou W, Xiao Z, Wen X, Luo J, Chen S, Cheng Z, et al. The anti-inflammatory effect of Andrographis paniculata (Burm. f.) Nees on pelvic inflammatory disease in rats through down-regulation of the NF-kappaB pathway. BMC Complement Altern Med 2016; 16:483-489.
11. Nanda A, K T, Banerjee P, Dutta M, Wangdi T, Sharma P, et al. Cytokines, angiogenesis, and extracellular matrix degradation are augmented by oxidative stress in endometriosis. Ann Lab Med 2020; 40:390-397.
12. Wen Y, Chen H, Luo F, Zhao L, Shu M, Su S, et al. Chlamydia trachomatis Plasmid Protein pORF5 Up-Regulates ZFAS1 to Promote Host Cell Survival via MAPK/p38 Pathway. Front Microbiol 2020; 11:11:593295-593305.
13. Bora G, Yaba A. The role of mitogen-activated protein kinase signaling pathway in endometriosis. J Obstet Gynaecol Res 2021; 47:1610-1623.
14. Ohman H, Tiitinen A, Halttunen M, Lehtinen M, Paavonen J, Surcel HM. Cytokine polymorphisms and severity of tubal damage in women with Chlamydia-associated infertility. J Infect Dis 2009; 199:1353-1359.
15. Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: At the crossroads of cell signalling and inflammatory disease. Biochim Biophys Acta 2014; 1843:2563-2582.
16. Li Z, Jin H, Yan Q, Sun L, Wasan HS, Shen M, et al. The method of activating blood and dredging collaterals for reducing chemotherapy-induced peripheral neuropathy: A systematic review and meta-analysis. Evid Based Complement Alternat Med 2019; 2019:1029626-1029643.
17. Hoffman RD, Li CY, He K, Wu X, He BC, He TC, et al. Chinese herbal medicine and its regulatory effects on tumor related T cells. Front Pharmacol 2020; 11:492-505.
18. Cai Y, Liang Q, Chen W, Chen M, Chen R, Zhang Y, et al. Evaluation of HuoXueHuaYu therapy for nonalcoholic fatty liver disease: A systematic review and meta-analysis of randomized controlled trial. BMC Complement Altern Med 2019; 19:178-188.
19. Liu SH, Chen PS, Huang CC, Hung YT, Lee MY, Lin WH, et al. Unlocking the mystery of the therapeutic effects of chinese medicine on cancer. Front Pharmacol 2020; 11:601785-601793.
20. Yuan S, Gong Y, Chen R, Du J, Zhang H, Chen T. Chinese herbal formula QHF inhibits hepatocellular carcinoma metastasis via HGF/c-Met signaling pathway. Biomed Pharmacother 2020; 132:110867.
21. Chen J, Qiu M, Li Y, Zhang Q, Zhang Y, Lin S, et al. Effects of Huatan Tongluo decoction on vascular endothelial growth factor receptor 2 expression in synovial tissues of rats with collagen-induced arthritis. J Tradit Chin Med 2019; 39:191-198.
22. Han J, Zhang Y, Shi X, Peng Z, Xing Y, Pang X. Tongluo Digui decoction treats renal injury in diabetic rats by promoting autophagy of podocytes. J Tradit Chin Med 2021; 41:125-132.
23. Li X. Tongluo decoction blocks apoptotic mitochondrial pathway and inhibits cardiomyocyte apoptosis of coronary microembolization rats. J Complement Altern Med 2020; 2:1-9.
24. Zhang F, Zhai M, Wu Q, Jia X, Wang Y, Wang N. Protective effect of tong-qiao-huo-xue decoction on inflammatory injury caused by intestinal microbial disorders in stroke rats. Biol Pharm Bull 2020; 43:788-800.
25. Liu S-H, Chen P-S, Huang C-C, Hung Y-T, Lee M-Y, Lin W-H, et al. Unlocking the mystery of the therapeutic effects of chinese medicine on cancer. Front Pharmacol 2021; 11:60178-60186.
26. Liu C, Qiu H, Huang R, Chai H, Yuan G, Shan S. Therapeutic mechanism and clinical observation of traditional chinese medicine combined with interventional recanalization for tubal infertility. Evid Based Complement Alternat Med 2021; 2021:2842250-2842259.
27. Huang C, He X, Luo W, Chen H, Huang Y. Combined chitosan and Dan-shen injection for long-term tubal patency in fallopian tube recanalization for infertility. Drug Deliv Transl Res 2019; 9:738-747.
28. Huang C, Wu Q, Liang J, Wang Q, He X, Xie Y, et al. Dose-effect relationship of chitosan and danshen combined injection for fallopian tube recanalization. Front Pharmacol 2022; 13:935117-935127.
29. Páez-Canro C, Alzate JP, González LM, Rubio-Romero JA, Lethaby A, Gaitán HG. Antibiotics for treating urogenital Chlamydia trachomatis infection in men and non-pregnant women. Cochrane Database Syst Rev 2019; 1:Cd010871-10941.
30. Derbie A, Mekonnen D, Woldeamanuel Y, Abebe T. Azithromycin resistant gonococci: A literature review. Antimicrob Resist Infect Control 2020; 9:138-144.
31. Liang Y, Xu K, Zhang P, Zhang J, Chen P, He J, et al. Quercetin reduces tendon adhesion in rat through suppression of oxidative stress. BMC Musculoskelet Disord 2020; 21:608-613.
32. Chen R, Xiao Y, Chen M, He J, Huang M, Hong X, et al. A traditional Chinese medicine therapy for coronary heart disease after percutaneous coronary intervention: A meta-analysis of randomized, double-blind, placebo-controlled trials. Biosci Rep 2018; 38:BSR20180973.
33. Wang SS, Jing-Wen HU, Sun HX, Hospital NC. Clinic Observation of Huoxue Tongluo Decoction Cooperated Hydrotubation in Tubal Infertility Treatment. Zhongguo Shi Yan Fang Ji Xue Za Zhi 2016;6:66.
34. Lemoine L, Thijssen E, Carleer R, Cops J, Lemmens V, Eyken PV, et al. Body surface area-based versus concentration-based intraperitoneal perioperative chemotherapy in a rat model of colorectal peritoneal surface malignancy: Pharmacologic guidance towards standardization. Oncotarget 2019; 10:1407-1424.
35. Ru J, Li P, Wang J, Zhou W, Li B, Huang C, et al. TCMSP: A database of systems pharmacology for drug discovery from herbal medicines. J Cheminform 2014; 6:13-18.
36. Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, et al. STRING v11: Protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res 2019; 47:D607-d613.
37. Doncheva NT, Morris JH, Gorodkin J, Jensen LJ. Cytoscape stringapp: Network analysis and visualization of proteomics data. J Proteome Res 2019; 18:623-632.
38. Liu J, Liu J, Tong X, Peng W, Wei S, Sun T, et al. Network pharmacology prediction and molecular docking-based strategy to discover the potential pharmacological mechanism of huai hua san against ulcerative colitis. Drug Des Devel Ther 2021; 15:3255-3276.
39. Brogden RN, Heel RC, Speight TM, Avery GS. Cefoxitin: A review of its antibacterial activity, pharmacological properties and therapeutic use. Drugs 1979; 17:1-37.
40. Cyranoski D. Why chinese medicine is heading for clinics around the world. Nature 2018; 561:448-450.
41. Lam TP. Strengths and weaknesses of traditional Chinese medicine and Western medicine in the eyes of some Hong Kong Chinese. J Epidemiol Community Health 2001; 55:762-765.
42. Gao AS, Du H, Gao Q, Wang YJ, Wang XY, Liu JW, et al. Tong luo jiu nao, a chinese medicine formula, reduces inflammatory stress in a mouse model of alzheimer’s disease. Am J Alzheimers Dis Other Demen 2016; 31:413-421.
43. Kany S, Vollrath JT, Relja B. Cytokines in inflammatory disease. Int J Mol Sci 2019; 20.
44. Lacy P, Stow JL. Cytokine release from innate immune cells: Association with diverse membrane trafficking pathways. Blood 2011; 118:9-18.
45. Kataoka T. Chemical biology of inflammatory cytokine signaling. J Antibiot (Tokyo) 2009; 62:655-667.
46. Schuster S, Cabrera D, Arrese M, Feldstein AE. Triggering and resolution of inflammation in NASH. Nat Rev Gastroenterol Hepatol 2018; 15:349-364.
47. Uciechowski P, Dempke WCM. Interleukin-6: A Masterplayer in the Cytokine Network. Oncology 2020; 98:131-137.
48. Fiebelkow J, Guendel A, Guendel B, Mehwald N, Jetka T, Komorowski M, et al. The tyrosine phosphatase SHP2 increases robustness and information transfer within IL-6-induced JAK/STAT signalling. Cell Commun Signal 2021; 19:94-112.
49. Banerjee S, Biehl A, Gadina M, Hasni S, Schwartz DM. JAK-STAT signaling as a target for inflammatory and autoimmune diseases: Current and future prospects. Drugs 2017; 77:521-546.
50. Basu A, Das AS, Borah PK, Duary RK, Mukhopadhyay R. Biochanin A impedes STAT3 activation by upregulating p38δ MAPK phosphorylation in IL-6-stimulated macrophages. Inflamm Res 2020; 69:1143-1156.
51. Cargnello M, Roux PP. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev 2011; 75:50-83.
52. Kaur S, Bansal Y, Kumar R, Bansal G. A panoramic review of IL-6: Structure, pathophysiological roles and inhibitors. Bioorg Med Chem 2020; 28:115327.
53. Abualsunun WA, Sahin C, Cummins CL, Piquette-Miller M. Essential role of STAT-3 dependent NF-κB activation on IL-6-mediated downregulation of hepatic transporters. Eur J Pharm Sci 2020; 143:105151.
54. Michaud-Levesque J, Bousquet-Gagnon N, Béliveau R. Quercetin abrogates IL-6/STAT3 signaling and inhibits glioblastoma cell line growth and migration. Exp Cell Res 2012; 318:925-935.
55. Kim HI, Lee SJ, Choi YJ, Kim MJ, Kim TY, Ko SG. Quercetin induces apoptosis in glioblastoma cells by suppressing Axl/IL-6/STAT3 Signaling Pathway. Am J Chin Med 2021; 49:767-784.