Fundamental Study On Substantial Basis Of Anti-gastric Cancer Activity Of Compound Xiao-ai-fei Mi-gao

Objective: To investigate antitumor activity in vitro, inhibitory activity on gastriccancer in vivo, anti-inflammatory activity in vivo and antioxidant activity in vitro,chemical compositions, extraction process of traditional Uyghur medicine-Fufangxiao-ai-fei migao (Co. XAFMG) based on it’s clinical use, for providing substantialfoudation for its secondary development. Methods:1) MFC transplanted tumor on ICRmice model was established to evaluate the in vivo therapeutical efficiency of ethanolextract of Co. XAFMG preparation. The expression of Proliferating Cell Nuclear Antigen(PCNA), vascular endothelial growth factor (VEGF) and CD31were detected byimmuno-histochemistry;2) Auricle edema induced by dimethylbenzene in mice, edema infeet induced by carrageen, cotton pellet granuloma and pleurisy model induced bycarrageen in rat were used for anti-inflammatory experiment. Content of protein, PGE2and TNF-α were determined;3) MTT assay was used to examine the antitumor activity invitro, by using gastric cancer BGC-823cells.The anticancer activities of differentabstraction parts of Co. XAFMG ethanol extract were screened;4) The antioxidantproperties of different parts of Co. XAFMG ethanol extract were evaluated by differentantioxidant tests in vitro, including diphenyl-2-bitter hydrazo (DPPH·) free radical,hydroxyl free radical (·OH), super oxide anion (O2-·) free radical scavenging ability, andiron ion reduction activity;5) GC/MS was uesed to analyze the essential oils constituentsof Co. XAFMG; silica gel column choromatography, macroporous absorbent resinchoromatography, polyamide column choromatography, sephadex LH-20columnchoromatography were used to isolate the chemical compounds;1H-NMR,13C-NMR,EI/MS were used to identify the chemical structes;6) The orthogonal experimental designmethods were used to optimize the extraction efficiency of effective part, by using theextraction ration of Piperine Galangin and extract yield as index, and their contents wereby UV-visible spectrophotometer; and the single factors as solid-liquid ratio, extraction temperature and extraction time influencing alkaloid and flavonoid contents wereinvestigated. Results:1) Therapeatic trial of MFC in tumor-bearing mice showed thatethanol extract of Co. XAFMG preparation obviously inhibited the growth of MFC tumorin a dose-dependent manner. The tumor weight inhibitory rate of ethanol extract of Co.XAFMG (0.034g/kg,0.068g/kg,0.136g/kg) ware55.2%,68.4%,59.9%, respectively.Data of immuno-histochemistry indicated that the expression of PCNA, VEGF and CD31all decreased after treatment with ethanol extract of Co. XAFMG preparation (0.068g/kg,0.136g/kg), in a dose-dependent manner;2) Co. XAFMG preparation has ananti-inflammatory effect, the effective dose range is0.034～0.136g/Kg. It could inhibitauricular edema induced by dimethylbenzene in mice, Carrageenin-induced inflammationin rats and granulation induced by cotton ball in rats, and reduce the content of protein inthe extravasate, PGE2and TNF-α in blood from rat pleurisy in dose-dependent manner;3)The petroleum ether part, chloroform part from ethanol extract and the essential oil of Co.XAFMG were showed considerable inhibitory effect on gastric cancer BGC-823cells invitro. Tumor cell viability of petroleum ether part were55.41%,25.32%,19.9%(50μg/mL,100μg/mL,500μg/mL); Tumor cell viability of chloroform part part were22.21%,21.70%(100μg/mL,500μg/mL); Tumor cell viability of the essential oil were26.91%,14.54%,15.65%,26.73%(1μg/mL,50μg/mL,100μg/mL,500μg/mL) Tumor cell viability ofGalangin and Piperine were27.42%、52.33%(500μg/mL);4) The chloroform part andn-butanol part of Co. XAFMG ethanol ectract showed relatively strong antioxidantactivity; however, its petroliu part and water part not showed this activity;5)28compounds were identified from essential oil,56compounds were identified frompetroleum ether extract,5compouds from petroleum ether part,7compouds fromchloroform part,2compouds from n-butanol part of ethanol extract were isolated andidentified;6) Optimum extraction technology was as follows: extracted2times (3h pertime) with30times of ethanol amount at70℃. Conclusion:1) The ethanol extract ofCo. XAFMG possess therapeutic action on MFC transplanted tumor-bearing mice, and it’spossible mechanism of actions involved in the down-regulation of the protein expressionof PCNA, VEGF and CD31;2) Co. XAFMG has anti-inflammatory effect by inhibitingthe production of PGE2and TNF-α in blood;3) The essential oil, petroleum etherextraction and chloroform extraction from the ethanol extract of Co. XAFMG preparation,Galangin and Piperine were initially identified as the active part of this preparation;4) Theeffective constituents with free radical scavenging activity of Co. XAFMG preparationwere distributed on the chloroform extract and the butanol extract of this preparation;5) The small molecule active compounds from Co. XAFMG were alkaloids, flavonoids,diphenyl-heptanes and essential oil;6) This optimized extraction technology was simple,stable and feasible, which could provide experimental basis for developing of Co.XAFMG.