Chinese Journal of Experimental Traditional Medical Formulae
Modified Wenjingtang Affects Caspase-8 MMP-9 N-cadherin and E-cadherin in Local Microenvironment of EM Rats with Kidney Deficiency and Blood Stasis
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Shanxi University of Chinese Medicine,Jinzhong 030619,China

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    Abstract:

    Objective To investigate the therapeutic effect and mechanism of modified Wenjingtang on endometriosis (EM) rats with kidney deficiency and blood stasis.Method Healthy non-pregnant female Sprague-Dawley (SD) rats of SPF grade were randomly divided into the blank group and experimental group. After being modeled via soaking in ice water and subcutaneous injection of epinephrine hydrochloride, the ones in the experimental group were further divided into the sham operation group and EM model group, with the former only undergoing laparotomy and the latter further receiving autologous endometrial transplant for triggering EM. The successfully modeled rats with EM due to kidney deficiency and blood stasis were randomized into the positive drug (danazol, 63 mg·kg-1) group and low- (5 g·kg-1), medium- (10 g·kg-1), and high-dose (20 g·kg-1) modified Wenjingtang groups. The corresponding drugs were administered by gavage, once per day, for four weeks. Then the ectopic and eutopic endometrial tissues were stained with hematoxylin-eosin (HE) to observe the histopathological changes. The protein and mRNA expression levels of cysteinyl aspartate-specific proteinase-8 (Caspase-8), matrix metalloproteinase-9 (MMP-9), N-cadherin, and E-cadherin were detected by immunohistochemistry (IHC), Western blotting, and real-time polymerase chain reaction (Real-time PCR), respectively.Result The IHC and Western blot revealed that the protein expression levels of MMP-9 and N-cadherin in eutopic and ectopic endometrial tissues of the model group were significantly increased as compared with those in the sham operation group (P<0.01), while the levels of Caspase-8 and E-cadherin was significantly decreased (P<0.01). Compared with the model group, the danazol and low-, medium-, and high-dose modified Wenjingtang groups exhibited obviously down-regulated MMP-9 and N-cadherin protein expression in eutopic and ectopic endometrial tissues (P<0.05,P<0.01), but up-regulated Caspase-8 and E-cadherin (P<0.05, P<0.01). Real-time PCR uncovered that the mRNA expression levels of MMP-9 and N-cadherin in eutopic and ectopic endometrial tissues of the model group were significantly elevated as compared with those in the sham operation group (P<0.01), whereas the levels of Caspase-8 and E-cadherin significantly declined (P<0.01). The comparison with the eutopic endometrial tissue in the model group showed that the mRNA expression levels of MMP-9 and N-cadherin in the danazol group and high- and medium-dose modified Wenjingtang groups were significantly down-regulated, while those of Caspase-8 and E-cadherin were significantly up-regulated (P<0.05,P<0.01).Conclusion Modified Wenjingtang alleviates the immunosuppression and blocks the angiogenesis in EM rats with kidney deficiency and blood stasis syndrome by regulating the expression of such cytokines as Caspase-8, MMP-9, N-cadherin, and E-cadherin, thus exerting the therapeutic effect against EM. The above-mentioned micro-indicators are potential markers reflecting the disease (EM), syndrome (kidney deficiency and blood stasis), and pathological mechanisms (immunosuppression and angiogenesis).

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History
  • Received:February 23,2021
  • Revised:
  • Adopted:
  • Online: May 06,2021
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