The Effect On Growth Of Astrocytes With Zuogui Wan And Yougui Wan In Vitro

Objective:This study is to observe the isolation, cultivation, purification and identification of astrocytes which is rat cerebral cortexcortical, observation of effect which clissical Chinese medicine (Zuogui Wan and Yougui Wan) have impacts on astrocytes in vitro.Method:1. We select the SD rats, and administer intragastrically with different concentrations liquid of Zuogui Wan and Yougui Wan,blood was collected from the aorta abdominalis to get drug-serum; 2. The brain cortex was isolated from the cerebral cortex of SD rats, and the cells were cultured, purified and identified by immunohistochemistry;3. Culturing astrocytes in culture medium which contains the different concentrations of Zuogui Wan and Yougui Wan serum.After 12 hours,we detect drug toxicity and determine the optimal dosing time (24h,48h,72h)by MTT, and test the content of brain derived neurotrophic factor (BDNF) with ELISA method. It is to illustrate influence on different concentrations of Zuogui Wan and Yougui Wan on astrocytes in vitro.Results:1. We got P3 cells which growth morphology, structural stability,through primary and culture and identification.It is used in the following experiments.2. In the cytotoxicity test,different concentrations of Zuogui Wan, Yougui Wan medicated serum on astrocytes, observed 0.92g/ml group of cells massively disaggregated and died, showed significant contraction of the 0.46g/ml group of cells, even necrosis, It showed that drug serum of higher concentration has obvious toxic effects on cell, according to the MTT test results.Generally speaking, inhibiting concentration is about 0.92g/ml,0.46g/ml on cells; and lower concentrations of drugs that promote cell proliferation is 0.12g/ml,0.23g/ml with the control group difference was significant (P< 0.05). Therefore, the determination of the drug concentration in cell proliferation assays were:0.12g/ml,0.23g/ml.3. MTT test results of cells proliferation assay show that the concentration (0.12g/ml,0.23 g/ml) zuogui Wan and Yougui Wan medicated serum at 24h,48h proliferation has obvious inpromoting, and compared to the control group,and had statistical significance (P<0.05),but at 72 hours of cells inrecase proliferation have no significant difference (P> 0.05)compared with control group.4. ELISA method was used to detect BDNF of astrocytes secreted have found that the drug concentration (0.12g/ml,0.23g/ml) effects on astroglial cells at 24h,48h have difference (P< 0.05) compared with the control group,compared difference (P< 0.05) and had significant difference (P<0.01)at 48h.Conclusions:There have different effects that different concentrations of Zuogui Wan and Yougui Wan medicated serum on astrocytes in vitro, higher concentrations (0.92g/ml,0.46g/ml) produce toxicity on astrocytes, and lower concentration (0.12g/ml,0.23g/ml) have role in promoting proliferation on astrocytes.