Platelets play an important role in the development of endometriosis. Scutellarin is a flavonoid isolated from a medicinal herb traditionally used as a potent antiplatelet agent. In this study, we sought to evaluate its potential therapeutic effect, if any, in mice with induced endometriosis. Endometriosis was induced in 27 female Balb/c mice by intraperitoneal injection of uterine fragments. Two weeks after the induction, the 27 mice were randomly divided in equal sizes into 3 groups: untreated, which received only vehicle, and low-dose and high-dose groups, which received low- and high dose of scutellarin treatment. Hotplate test was administrated to all mice before endometriosis induction, and before and after the scutellarin treatment. Two weeks after the treatment, a blood sample was drawn before sacrifice and all lesions were harvested. The peripheral platelet activation rate and total lesion weight were assessed, and immunohistochemistry and histochemistry analyses were performed to evaluate the extent of proliferation, angiogenesis, fibroblast-to-myofibroblast transdifferentiation (FMT), and fibrosis in lesions. Compared with untreated mice, mice in both low-dose and high-dose groups had significantly reduced lesion weight and improved hyperalgesia. Scutellarin also reduced the peripheral-activated platelets rate and resulted in significantly reduced platelet aggregation, cellular proliferation, angiogenesis, the extent of FMT, and the extent of fibrosis in lesions. Thus, we conclude that scutellarin is efficacious in treating endometriosis in vivo by suppressing platelet aggregation, inhibiting proliferation, angiogenesis, and fibrogenesis, resulting in reduced lesion size and improved pain behavior. As such, scutellarin may be a potentially promising therapeutics for the treatment of endometriosis.
Practice Committee of the American Society for Reproductive Medicine. Treatment of pelvic pain associated with endometriosis: a committee opinion. Fertil Steril. 2014;101(4):927–935.
3.
GuoSW. Recurrence of endometriosis and its control. Hum Reprod Update. 2009;15(4):441–461.
4.
KogaKTakamuraMFujiiTOsugaY. Prevention of the recurrence of symptom and lesions after conservative surgery for endometriosis. Fertil Steril. 2015;104(4):793–801.
5.
Garcia-VelascoJASomiglianaE. Management of endometriomas in women requiring IVF: to touch or not to touch. Hum Reprod. 2009;24(3):496–501.
6.
de ZieglerDBorgheseBChapronC. Endometriosis and infertility: pathophysiology and management. Lancet. 2010;376(9742):730–738.
7.
CocciaMERizzelloFMarianiGBullettiCPalagianoAScarselliG. Ovarian surgery for bilateral endometriomas influences age at menopause. Hum Reprod. 2011;26(11):3000–3007.
8.
VercelliniPBuggioLFrattaruoloMPBorghiADridiDSomiglianaE. Medical treatment of endometriosis-related pain. Best Pract Res Clin Obstet Gynaecol. 2018;51:68–91.
9.
VercelliniPCrosignaniPSomiglianaEViganoPFrattaruoloMPFedeleL. ‘Waiting for Godot’: a commonsense approach to the medical treatment of endometriosis. Hum Reprod. 2011;26(1):3–13.
10.
GuoSWGroothuisPG. Is it time for a paradigm shift in drug research and development in endometriosis/adenomyosis?Hum Reprod Update. 2018;24(5):577–598.
11.
BrosensIA. Endometriosis – a disease because it is characterized by bleeding. Am J Obstet Gynecol. 1997;176(2):263–267.
12.
GuoSWDingDShenMLiuX. Dating endometriotic ovarian cysts based on the content of cyst fluid and its potential clinical implications. Reprod Sci. 2015;22(7):873–883.
13.
DingDLiuXDuanJGuoSW. Platelets are an unindicted culprit in the development of endometriosis: clinical and experimental evidence. Hum Reprod. 2015;30(4):812–832.
14.
GuoSW. Fibrogenesis resulting from cyclic bleeding: the Holy Grail of the natural history of ectopic endometrium. Hum Reprod. 2018;33(3):353–356.
15.
ZhangQDuanJLiuXGuoSW. Platelets drive smooth muscle metaplasia and fibrogenesis in endometriosis through epithelial–mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation. Mol Cell Endocrinol. 2016;428:1–16.
16.
ZhangQDuanJOlsonMFazleabasAGuoSW. Cellular changes consistent with epithelial–mesenchymal transition and fibroblast-to-myofibroblast transdifferentiation in the progression of experimental endometriosis in baboons. Reprod Sci. 2016;23(10):1409–1421.
DuYLiuXGuoSW. Platelets impair natural killer cell reactivity and function in endometriosis through multiple mechanisms. Hum Reprod. 2017;32(4):794–810.
19.
WuQDingDLiuXGuoSW. Evidence for a hypercoagulable state in women with ovarian endometriomas. Reprod Sci. 2015;22(9):1107–1114.
20.
DingDLiuXGuoSW. Further evidence for hypercoagulability in women with ovarian endometriomas. Reprod Sci. 2018;25(11):1540–1548. doi:10.1177/1933719118799195
21.
GuoSWDingDGengJGWangLLiuX. P-selectin as a potential therapeutic target for endometriosis. Fertil Steril. 2015;103(4):990–1000e8.
22.
GuoSWDingDLiuX. Anti-platelet therapy is efficacious in treating endometriosis induced in mouse. Reprod Biomed Online. 2016;33(4):484–499.
23.
CarneroA. High throughput screening in drug discovery. Clin Transl Oncol. 2006;8(7):482–490.
24.
WangLMaQ. Clinical benefits and pharmacology of scutellarin: a comprehensive review. Pharm Ther. 2018;190:105–127.
25.
LiuXChengJZhangG, et al.Engineering yeast for the production of breviscapine by genomic analysis and synthetic biology approaches. Nat Commun. 2018;9(1):448.
26.
LuoPTanZHZhangZFZhangHLiuXFMoZJ. Scutellarin isolated from Erigeron multiradiatus inhibits high glucose-mediated vascular inflammation. Yakugaku Zasshi. 2008;128(9):1293–1299.
27.
MaJYJiangWWZhouZTLiJMWangHY. The promoting angiogenesis and anti-inflammation effect of scutellarin on polyglycolic acid scaffold of Balb/c mice model. J Asian Nat Prod Res. 2008;10(11-12):1147–1153.
28.
TianXChangLMaG, et al.Delineation of platelet activation pathway of scutellarein revealed its intracellular target as protein kinase C. Biol Pharm Bullet. 2016;39(2):181–191.
29.
HouLChenLFangL. Scutellarin inhibits proliferation, invasion, and tumorigenicity in human breast cancer cells by regulating HIPPO-YAP signaling pathway. Med Sci Monit. 2017;23:5130–5138.
30.
KeYBaoTWuX, et al.Scutellarin suppresses migration and invasion of human hepatocellular carcinoma by inhibiting the STAT3/Girdin/Akt activity. Biochem Biophys Res Commun. 2017;483(1):509–515.
31.
LiHHuangDGaoZChenYZhangLZhengJ. Scutellarin inhibits the growth and invasion of human tongue squamous carcinoma through the inhibition of matrix metalloproteinase-2 and -9 and alphavbeta6 integrin. Int J Oncol. 2013;42(5):1674–1681.
32.
ZhuPTMaoMLiuZGTaoLYanBC. Scutellarin suppresses human colorectal cancer metastasis and angiogenesis by targeting ephrinb2. Am J Transl Res. 2017;9(11):5094–104.
33.
SunCYZhuYLiXF, et al.Scutellarin increases cisplatin-induced apoptosis and autophagy to overcome cisplatin resistance in non-small cell lung cancer via ERK/p53 and c-met/AKT signaling pathways. Front Pharmacol. 2018;9:92.
34.
PanZZhaoWZhangX, et al.Scutellarin alleviates interstitial fibrosis and cardiac dysfunction of infarct rats by inhibiting TGFbeta1 expression and activation of p38-MAPK and ERK1/2. Br J Pharmacol. 2011;162(3):688–700.
35.
LiYLiuYDChenSL, et al.Down-regulation of long non-coding RNA MALAT1 inhibits granulosa cell proliferation in endometriosis by up-regulating P21 via activation of the ERK/MAPK pathway. Mol Hum Reprod. 2018.
36.
KhoshdelRadNSalehiZMashayekhiFAbbasiOMirzajaniE. Soluble c-Met expression in the peritoneal fluid and serum of patients with different stages of endometriosis. Arch Gynecol Obstet. 2014;289(5):1107–1112.
37.
ChoiJJoMLeeEHwangSChoiD. Aberrant PTEN expression in response to progesterone reduces endometriotic stromal cell apoptosis. Reproduction. 2017;153(1):11–21.
38.
Council NR. Guide for the Care and Use of Laboratory Animals. Washington, DC: National Academies Press; 1996.
39.
SomiglianaEViganoPRossiGCarinelliSVignaliMPanina-BordignonP. Endometrial ability to implant in ectopic sites can be prevented by interleukin-12 in a murine model of endometriosis. Hum Reprod. 1999;14(12):2944–2950.
40.
LongQLiuXGuoSW. Surgery accelerates the development of endometriosis in mouse. Am J Obstetr Gynecol. 2016;215(3):320.e1–320.e15.
41.
Reagan-ShawSNihalMAhmadN. Dose translation from animal to human studies revisited. FASEB J. 2008;22(3):659–661.
42.
HuXTengSHeJ, et al.Pharmacological basis for application of scutellarin in Alzheimer’s disease: antioxidation and antiapoptosis. Mol Med Rep. 2018;18(5):4289–4296.
43.
SunCLiCLiX, et al.Scutellarin induces apoptosis and autophagy in NSCLC cells through ERK1/2 and AKT signaling pathways in vitro and in vivo. J Cancer. 2018;9(18):3247–3256.
44.
LuYNieJLiuXZhengYGuoSW. Trichostatin A, a histone deacetylase inhibitor, reduces lesion growth and hyperalgesia in experimentally induced endometriosis in mice. Hum Reprod. 2010;25(4):1014–1025.
45.
R Development Core Team. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing; 2016.
46.
ZhaoTLiuXZhenXGuoSW. Levo-tetrahydropalmatine retards the growth of ectopic endometrial implants and alleviates generalized hyperalgesia in experimentally induced endometriosis in rats. Reprod Sci. 2011;18(1):28–45.
47.
ZhengYLiuXGuoSW. Therapeutic potential of andrographolide for treating endometriosis. Hum Reprod. 2012;27(5):1300–1313.
48.
LiuMLiuXZhangYGuoSW. Valproic acid and progestin inhibit lesion growth and reduce hyperalgesia in experimentally induced endometriosis in rats. Reprod Sci. 2012;19(4):360–373.
49.
LienLMSuCCHsuWH, et al.Mechanisms of andrographolide-induced platelet apoptosis in human platelets: regulatory roles of the extrinsic apoptotic pathway. Phytother Res. 2013;27(11):1671–1677.
50.
DavidsonDCHirschmanMPSpinelliSL, et al.Antiplatelet activity of valproic acid contributes to decreased soluble CD40 ligand production in HIV type 1-infected individuals. J Immunol. 2011;186(1):584–591.
51.
NurdenATNurdenPSanchezMAndiaIAnituaE. Platelets and wound healing. Front Biosci. 2008;13:3532–3548.
52.
AssoianRKKomoriyaAMeyersCAMillerDMSpornMB. Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem. 1983;258:7155–7160.
HuXMZhouMMHuXMZengFD. Neuroprotective effects of scutellarin on rat neuronal damage induced by cerebral ischemia/reperfusion. Acta Pharmacol Sin. 2005;26(12):1454–1459.
55.
YangWLustRMBofferdingAWingardCJ. Nitric oxide and catalase-sensitive relaxation by scutellarin in the mouse thoracic aorta. J Cardiovasc Pharmacol. 2009;53(1):66–76.
56.
Gonzalez-RamosRDonnezJDefrereS, et al.Nuclear factor-kappa B is constitutively activated in peritoneal endometriosis. Mol Hum Reprod. 2007;13(7):503–509.
57.
Gonzalez-RamosRVan LangendoncktADefrereS, et al.Involvement of the nuclear factor-kappaB pathway in the pathogenesis of endometriosis. Fertil Steril. 2010;94(6):1985–1994.
58.
KaponisAIwabeTTaniguchiF, et al.The role of NF-kappaB in endometriosis. Front Biosci (Schol Ed). 2012;4:1213–1234.
59.
WangYHGengLLiH. Study on effect of scutellarin in resisting liver fibrosis in rats [in Chinese]. Zhongguo Zhong Yao Za Zhi. 2015;40(10):1999–2003.
60.
ZhouHChenXChenL, et al.Anti-fibrosis effect of scutellarin via inhibition of endothelial–mesenchymal transition on isoprenaline-induced myocardial fibrosis in rats. Molecules. 2014;19(10):15611–15623.
61.
WipffPJRifkinDBMeisterJJHinzB. Myofibroblast contraction activates latent TGF-beta1 from the extracellular matrix. J Cell Biol. 2007;179(6):1311–1323.
62.
FroeseARShimboriCBellayePS, et al.Stretch-induced activation of transforming growth factor-beta1 in pulmonary fibrosis. Am J Respir Crit Care Med. 2016;194(1):84–96.
63.
MatsuzakiSCanisMPoulyJLDarchaC. Soft matrices inhibit cell proliferation and inactivate the fibrotic phenotype of deep endometriotic stromal cells in vitro. Hum Reprod. 2016;31(3):541–553.
64.
MatsuzakiSDarchaCPoulyJLCanisM. Effects of matrix stiffness on epithelial to mesenchymal transition-like processes of endometrial epithelial cells: implications for the pathogenesis of endometriosis. Sci Rep. 2017;7:44616.
65.
MichelsonADBarnardMRKruegerLAFrelingerA IIIFurmanMI. Evaluation of platelet function by flow cytometry. Methods. 2000;21(3):259–270.
66.
ZhangQLiuXGuoSW. Progressive development of endometriosis and its hindrance by anti-platelet treatment in mice with induced endometriosis. Reprod Biomed Online. 2017;34(2):124–136.
67.
LiuXZhangQGuoSW. Histological and immunohistochemical characterization of the similarity and difference between ovarian endometriomas and deep infiltrating endometriosis. Reprod Sci. 2018;25(3):329–340.
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