| Poria cocos is a traditional Chinese medicine with the same origin as food and medicine,and has a reputation of ten medicines and nine poria cocos in ancient times.Modern research shows that its main active substance is poria cocos polysaccharide,which can inhibit the growth of cancer cells,inhibit inflammation,protect the liver,regulate the immune system,etc.However,poria cocos is mostly alkaline polysaccharide insoluble in water,which is difficult to play its active role.In order to solve this problem,Some studies obtained Carboxymethyl poria polysaccharide(CMP)with better solubility and higher bioactivity through structural modification of basic polysaccharide(carboxymethyl).However,CMP also needs to go through the declaration of new raw materials before going on the market,which has the disadvantages of long cycle and high cost.As to the previous research,this paper has obtained a Water-soluble poria polysaccharide(WSP)by enzymatic hydrolysis of alkaline poria polysaccharide and soluble soluble poria polysaccharide(WSP).Based on the above background,this project aims to explore the anti-inflammatory,skin damage repair and whitening effects of WSP.The main research results are as follows:(1)The results of CCK-8 cell safety test showed that 3,6,12,24,48,96,192 and 384μg/m L of WSP did not significantly inhibit the activity of RAW264.7 cells,indicating that WSP had good safety against RAW264.7 cells.The above concentration of WSP coincubated with LPS of 0.1 μg/m L did not significantly inhibit the activity of RAW264.7cells,suggesting that the above concentration of WSP could be used for subsequent cellbased anti-inflammatory tests.The anti-tumor results of Caco-2 cells treated with 50 μg/m L WSP for 12 h showed that the morphology of Caco-2 cells was significantly changed.Under the condition of 100μg/m L WSP,a small amount of Caco-2 cells began to die.Under the condition of 200μg/m L WSP,a large number of Caco-2 cells died and their contour disappeared.Under 400μg/m L WSP treatment,a large number of Caco-2 cells died and were suspended in cell medium.However,Caco-2 cells in blank group grew normally,indicating that WSP may have certain anticancer effect on Caco-2 cells.(2)The results of cell level anti-inflammatory test showed that the contents of IL-6 and TNF-α in RAW264.7 cells were significantly increased after LPS stimulation compared with blank group(p<0.001).Compared with the LPS group pretreated with WSP,6 μg/m L WSP significantly alleviated the increase of TNF-α and IL-1β contents induced by LPS(p<0.001),and the higher the concentration of WSP,the more significant the decrease of TNF-α and IL-1β contents,indicating that the anti-inflammatory effect of WSP was concentration-dependent.(3)The results of animal level anti-inflammatory test(mouse ear swelling test)showed that compared with blank group,the inhibition rate of ear swelling in hyaluronic acid group was-54.84%,that in CMP group was 10.32%,and that in WSP group was 22.12%.According to the above results,ear swelling of mice in CMP group and WSP group was significantly reduced(p<0.01),indicating that CMP and WSP have anti-inflammatory effects;In addition,the effect of WSP group on reducing xylene induced ear swelling was significantly better than that of CMP group(p<0.01).(4)The results of animal level skin injury repair test(rat skin alkali burn test)showed that the average recovery time of positive drug Jingwanhong group was 16 days,the average recovery time of CMP group was 19 days,and the average recovery time of WSP was 18 days,compared with 28 days in the model group.These results indicated that compared with model group,the recovery time of positive drug Jingwanhong group,CMP group and WSP group was significantly shortened,indicating that CMP and WSP could promote skin injury repair,but there was no significant difference between the two groups.(5)The results of anti-inflammatory tests at the animal level(mouse ulcerative colitis test)showed that WSP alleviated the apparent symptoms of ulcerative colitis: after ingestion of WSP and CMP,the trend of weight loss was inhibited,and DAI score of mice in the model group was significantly higher than that in the blank group(p<0.01);Compared with model group,DAI score in CMP group and WSP low-dose,medium-dose and high-dose groups was significantly decreased(p<0.01),there was no significant difference in DAI scores between the CMP group and the medium-dose group at the same dose.WSP can effectively improve the blood cell level of mice with ulcerative colitis,and the white blood cell level of mice ingestion of WSP and CMP is significantly lower than that of the DSS group(p<0.01),the monocyte level in CMP group and WSP low-dose,mediumdose and high-dose groups was significantly lower than that in DSS group(p<0.001),the erythrocyte count and HCT level in CMP and WSP medium dose groups were significantly higher than those in DSS group(p<0.01),the number of red blood cells and HCT level in low and high dose WSP groups were significantly higher than those in DSS group(p<0.001).WSP can repair tissue damage in ulcerative colitis,and colon length increased in CMP and WSP low-dose mice compared with model group(p<0.05),colon length of mice in medium and high dose groups of WSP was significantly increased(p<0.01);The pathological manifestations of colon tissue were also corresponding to the length of colon.The pathological conditions of colon in CMP group and WSP low-dose,medium-dose and high-dose groups were improved to varying degrees,and with the increase of WSP concentration,the damage of colon tissue would be alleviated.WSP can significantly reduce the levels of inflammatory factors in mice with ulcerative colitis.Compared with model group,the contents of TNF-α,IL-1β and IL-6 in serum of CMP group and WSP low-dose,medium-dose and high-dose groups were significantly reduced.(6)Whitening effect test(tyrosinase inhibition test)results showed that CMP and WSP had no inhibition effect on tyrosinase.In summary,this study found that WSP has significant anti-inflammatory effects on RAW264.7 inflammatory cells,skin inflammation and colitis,and can kill colon cancer cells Caco-2.In addition,this study also found that WSP can promote the repair of skin injuries. |
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