Polygala And Its Mechanism, ACROUSTATRINOWIISCHOTT Extract On PC12 Cell Processes Affected

1BackgroundWhether Synaptic plasticity can be improved or not is an important indicator of nerve injury disease, which includes structural plasticity and functional plasticity, and the former is the morphological basis of the latter. What’s more, growth and extending of neuron processes is not only an important symbol of synaptic structural plasticity, but also a direct behavior of synaptic plasticity in the cell level. In traditional Chinese medicine theory, Shenghui Decoction have nutritious effect on kidney and heart, was used to treat forgetfulness. Modem pharmacological studies found that it can improve the ability of learning and memory in mice. Preliminary results by our group found that Sihui Mixture can significantly improve learning and memory abilities of tired rats. To explore the molecular mechanisms of that, divided the Shenghui Decoction into four group, including Zibuganshen decoction (Radix Rehmanniae Preparata and Comus offcinalis), Ningxinanshen decoction (Spina Date Seed and Platycladus orientalis), Buqijianpi decoction (Poria cum Radix Pini and Radix Ginseng Rubra) and Qutankaiqiao decoction (Polygala tenuifolia Willd、Acorus tatarinowii and Semen sinapis), with Sini Power as the experiment drugs to detect their influence to cell processes growth. The results showed that Buqijianpi decoction and Qutankaiqiao decoction from Shenghui Decoctionhave the effect on promoting growth of cell processes, and can promote the expression level of GAP-43mRNA at the same time, which is closely related to neural plasticity. So from the perspective of synaptic plasticity, this experiment will use PC12cell as a model, growth of cell processes for the test indicator, to explore the influence of drugs on cell proliferation and differentiation, which may provide data to support further learning about identifying active ingredients of herbal to improve memory ability, and preliminary explore the possible mechanism of it.2MethodsThe first part, PC12cells was used as a model, and used western blot to detect the effect of Buqijianpi decoction and Qutankaiqiao decoction on expression level of the GAP-43, which is to explore the molecular mechanisms of drugs to cell neuritis outgrowth further.The second part, aqueous extract of Polygala tenuifolia Willd and Acorus tatarinowii were chosen as the test drugs, first we want to find the safe and effective concentration of the drugs in complete culture conditions, and to observe the effects of drugs on cells from the perspective of cell proliferation and cell differentiation respectively in low serum culture condition. In addition, whether drugs have the effect on neurotropic was detected by cck-8kit; on the other hand, observed cell morphology and made a statistics about cell processes length to found whether the drugs have an effects to promote cell differentiation.The third part, based on the experimental results of the second part, chose aqueous extract of Poly-gala tenuifolia Willd as the test drug. RT-PCR was used to detect the effects of drugs on expression of GAP-43gene level of PC12, while western blot was used for that of expression of GAP-43protein level.The fourth part, in reference to the traditional prescription about Polygala-Acorus tatarinowii proportion, used the length of cell processes for the detection of target, cell morphology and statistical analysis as methods, to explore the influence of different concentrations of the Polygala-Acorus tatarinowii drug to the length of cell processes with the same compatibility proportion, and in same concentration the influence of different proportion drugs to cell process growth.3Results1The decoction of compound medicine group (Polygala tenuifolia Willd、Acorus tatarinowii and Semen sinapis) significantly promoted the protein expression level of GAP-43in PC12cell.2Under complete culture conditions, aqueous extracts of Polygala tenuifolia Willd and Acorus tatarinowii could promote cell proliferation could significantly, and had no significant side effects.3Under low serum culture conditions, in group of Polygala tenuifolia Willd and Acorus tatarinowii, aqueous extract of Polygala tenuifolia Willd could significantly promote the effect of cell difienentiation in a concentration-dependent; aqueous extract of Acorus tatarinowii could significantly inhibit cell differentiation and promote cell proliferation at the same time,with a concentration dependent4Aqueous extract of Polygala tenuifolia Willd could significandy promote the expression of GAP-43in PC12cells, in both gene level and protein level.5The drug group of Polygala tenuifolia Willd and Acorus tatarinowii have an active effect on cell processes growth with a mixing ratio at1to4. A low concentration about10mg/l of drug group have the best effect on promoting the length of cell processes with a proportion of1:1; a relatively high concentration about200mg/l of drug group have the best effect on promoting the length of cell processes with a proportion of4:1, and have a inhibitory effect to cell processes growth with a proportion of1:1.