Analgesic Effect And Mechanism Study Of Natural Plant Antimicrobial Solution

Pain is a kind of disease with complex pathogenesis which can be quite harmful to people’s both physical and mental health, While most of the existing analgesic drugs have side effects such as tolerance, addiction and damaging organs.As topical linimentum derived from natural medicinal plants, Natural plant antimicrobial solution (PAMs) have various advantages, such as no n-tolerance, no n-addict ion, less adverse effects and so on.What’s more, PAMs own a terrific development prospect due to its multiple therapeutic effects: anti-inflammatory, analgesia,anti-microbial,anti-tumour as well as promoting wound healing and tissue renewal, which have been proved by early practical application of this product. Nevertheless, as traditional Chinese medicine compound preparation, PAMs’s unknown and complicated composition hindered the mechanism research of its efficacy, therefore,carrying on thequalitative and quantitative analysis of its active ingredients is of great importance for the follow-up research, safety evaluation and quality control.Firstly, this article employed flame atomic absorption spectrophotometry(FAAS) to determine heavy metals and trace elements in PAMs, the results showed that PAMs didn’t contain nickel(Ni), lead(Pb), chromium(Cr) and cadmium(Cd), but contain 0.0185μg·mL copper(Cu),6.13μg·gmL-1 iron (Fe)and 0.56μg·mL-1 zinc (Zn), and a small mount of Cu and abundant Fe and Zn could be beneficial to human body and PAMs’s medicinal effect. Secondly, high performance liquid chromatography (HPLC) was used to conduct thequalitative and quantitative analysis of some effective components in PAMs.The results showed that PAMs contained allantoin, hydroxyl safflower yellow pigment A, loureirin A, aconitine respectively at a concentration of 262.85μg/mL,94.235μg/mL,71.325μg/mL, 9.11μg/mL.Moreover, mice pain model induced by formaldehyde was used in this article to study the analgesic effect of PAMs.During the process of external inunctum, compared to test group(PAMs group), more and stronger resistance behavior was showd in positive control group, which indicated that PAMs was less irritating to the skin than the positive control drug.during two time phase:0-5min and 20-40min after subcutaneous injection of 5% formaldehyde solution in mice’s hindpaw, the pain scores of PAMs group were significantly lower than the negative control group(P<0.05),and higer than the positive control group(P>0.05), which indicated that PAMs could alleviate both the acute pain and formaldehyde induced peripheral inflammatory pain. What’s more, testing results of pain related factors in mice serum showed that PAMs and positive control group had lower concentration of PGE2than the negative control group(P<0.05),and their β-EP level were significantly higer than the negative control group(P<0.05), and the NO content of positive control group was obviously lower than that of negative control group(P<0.05), while PAMs and negative control group had no difference in the content of NO. Therefore, we inferred that PAMs could inhibit the synthesis and release of PGE2 and raise β-EP level to achieve its analgesia effect.Furthermore, this article established PC 12 cell damage model which was induced by Aβ25-35 to preliminary study the nerve cytoprotection effect of PAMs. Micoscopic observation and MTT test results demonstrated that PAMs at the concentration of 1%,0.5% and 0.25% had no inhibition for PC12’s normal growth. Pretreated with medicine for 2h, then treated with Aβ25-35,PC12 agglomerated and died because ofAβ25-35,which was observed by invert microscope. MTT and LDH test results showed that cell vitality of the model group was significantly lower than that of normal group (P<0.05), and its content of LDH was obviously higher than normal group(P<0.05), which confirmed the result of microscopic observation and demonstrated that modeling was successful. In addition, compared to model group,0.5%,0.25%of PAMs and 1% Hericiumerinaceus capsule increaseedcell vitality significantly(P<0.05) and reduced the content of LDH(P<0.05), which inferred that 0.5%,0.25% of PAMs, just like Hericiumerinaceus, could prevent and relieve oxidative damage caused by Aβ25-35.However, compared to model group, the group treated with1% PAMs showed obviously lower cell vitality((P<0.01) and lower content of LDH,which indicated that PAMs at a concentration of 1% had no effect in alleviating damage induced by Aβ25-35.Main conclusion:PAMs contain no nickel(Ni), lead(Pb), chromium(Cr) and cadmium(Cd),but contain 0.0185μg·mL-1 copper(Cu),6.13μg·mL-1 iron(Fe),0.56μg·mL-1 zinc (Zn).PAMs have abundant llantoin, hydroxyl safflower yellow pigment A, Loureirin A and a small number of aconitine. PAMs can inhibit the synthesis and release of PGE2 and raise β-EP level to achieve its anti-inflammatory analgesia effects, its low concentration can prevent and relive oxidative damage caused by Aβ25-35.