Effect Of Aesculetin On RBC Membrane Function In Tumor Bearing Mice

As a traditional medicine in China, the traditional Chinese medicin has a great advantage in terms of anti-tumor. Aesculetin (Aesculetin, AES) as coumarin is the main effective consti-tuents of Chinese medicine cortex Fraxini, has been reported that have anti-tumor activity both in vitro and in vivo, can inhibit the growth of tumor cells, and has immunomodulatory effects. Laboratory experiments have confirmed that AES can inhibit the growth of sarcoma S180 by immune mechanism, but there is no S180 in tumor bearing mice as animal model to explore the influence of AES on immune function of red blood cell (RBC) mechanisms. The RBC mem-brane is the material basis both of RBC immune anti-tumor and RBC carrier of anti-tumor, more novel RBC membrane drug discovery system, so how to combine the related mechanism of RBC immune with RBC membrane drug discovery system is of great value to study. This paper studies the effect of AES on the structure and composition of RBC membrane of tumor-bearing mice, in order to further reveal the mechanism of anti tumor immunity, provide a reference for future research.Objective:To investigate the effect of AES on RBC membrane in S180 tumor-bearing mice, revealing the red cell immune mechanism of anti-tumor, and provide a reference for the subse-quent in-depth study.Method:Modeling of tumor bearing mice by transplanted S180 sarcoma cells in plants. Male and female mice were randomly divided into 6 groups:normal control group (no S180 sarcoma cell line), model control group, positive control group (Astragalus polysaccharide, 100mg/kg), AES low dose group (20mg/kg), AES middle dose group (40mg/kg) and AES high dose group (80mg/kg). ①The RBC adhesion ability of tumor cells and RBC immune activity of positive and negative regulatory factors were tested by improved method of rosette test. ② The mobility of RBC membrane was tested by spectrophotometry with DPH labeled. ③The RBC membrane blocking degree was tested by UV spectrophotometry. ④The content of sialic acid (SA) and the ATP enzyme activity of RBC membrane were tested by UV spectrophoto-metry with kit. ⑤The RBC membrane potential was measured by flow cytometry. ⑥The expression of Band3 protein was measured by Western Blot. ⑦The anion transport activity of RBC membrane was measured by UV spectrophotometry.Results:Compared with the normal group, model group the rosette rate of RBC adhesion of tumor cells decreased to (11.05+3.05)%, suggesting that the RBC immune function was inhibited (P<0.01), and AES can recover the adhesion, having significant difference compared with the model group (P<0.01). The RFER and RFIR of the model group were (31.62±10.66)% and (59.21±7.05)%, compared with the normal group with very significant difference (P<0.01), suggesting that promoter activity was inhibited, inhibitor activity was promoted, immune regulation function imbalance, AES can increase promoting factor activity and reduce inhibiting factor activity to restore the RBC immune function of positive and negative regulation (P<0.05, P<0.01). Compared with the model group, the mobility of RBC membrane of AES middle dose and high dose group can be restored to 7.7373+2.5050 and 7.8757+1.7086, with very significant difference (P<0.01). RBC membrane blocking degree in the model group decreased to (32.07+7.47)%, compared with the normal group with very significant difference (P<0.01), given different concentrations of AES, the RBC membrane blocking degree in all dose group increased, compared with the model group with significant difference (P<0.05, P<0.01). The content of SA, ATPase and membrane potential of RBC membrane were suppressed, compared with the normal group with very significant difference (P<0.01); given different concentrations of AES, the three index all increased, compared with the model group, AES low dose group with no significant difference, middle and high dose group with very significant difference (P<0.01). The expression of Band3 protein and the anion transport activity of RBC membrane were all suppressed (P<0.01), given different concentrations of AES, the two index all enhanced, compared with the model group, all dose group with significant difference (P<0.05,P<0.01).Conclusion:AES can inhibit the S180 tumor-bearing mice tumor growth through the RBC immune function; it can enhance the RBC innate immune adhesion ability, and regulate the red cell immune activity of positive and negative regulatory factors so as to restore the balance. At the same time, the RBC membrane as the basic structure, its construction and composition is also affected, including the recovery of RBC membrane mobility and blocking degree, the raise of the content of SA, ATP enzyme activity and membrane potential level, the improvement of the expression of Band3 protein and anion transport the activity, to maintain the stability of RBC and ensure their normal activity, improving RBC innate immune adhesion ability, thereby enhancing the immune function of RBC and the anti-tumor effect.