RESUMEN
ObjectiveTo investigate the effects of Linggui Zhugantang （LGZGT）-containing serum on primary astrocytes （AS） induced by β amyloid 1-42 （Aβ1-42） in a rat model of Alzheimer's disease （AD） and explore the phagocytic and degradative effects of LGZGT on Aβ. MethodAn AD model was established by inducing AS with Aβ1-42. The cells were divided into normal group， model group， LGZGT low-， medium-， and high-dose （LGZGT-L， LGZGT-M， and LGZGT-H） groups， and donepezil hydrochloride group. The model group was treated with Aβ1-42 at a final concentration of 10 μmol∙L-1. The LGZGT-L， LGZGT-M， and LGZGT-H groups were treated with 10% serum containing LGZGT on the basis of the model group. Cell viability was assessed using a cell counting kit-8 （CCK-8）， lactate dehydrogenase （LDH） activity was measured using an LDH assay kit， and cell morphology was observed using an inverted microscope. The expression of Aβ-related degradation enzymes insulin-degrading enzyme （IDE） and cathepsin D （CTSD） was detected using Western blot， and the fluorescence intensity of cathepsin B （CTSB） was measured using immunofluorescence. The content of Aβ1-42 in cells was determined using an enzyme-linked immunosorbent assay （ELISA）. ResultCompared with the normal group， the viability of AS in all groups decreased， and Aβ1-42 at different concentrations had inhibitory effects on AS proliferation. After administration， compared with the normal group， the cell survival rate of the model group decreased significantly （P<0.05）. Compared with the model group， the cell survival rates of the LGZGT-H group and donepezil hydrochloride group increased significantly （P<0.05）. The LDH activity of cells in the model group was significantly increased compared with that in the normal group （P<0.05）， and cell bodies were swollen and enlarged with increased protrusions and elongation， suggesting more obvious cell damage. Compared with the model group， the LDH activity of cells in the donepezil hydrochloride， LGZGT-L， LGZGT-M， and LGZGT-H groups decreased significantly （P<0.05）. After administration， the cell swelling in the LGZGT-M， LGZGT-H， and donepezil hydrochloride groups improved， cell protrusions shortened， and cell clustering decreased. Compared with the normal group， the expression of IDE and CTSD in the model group decreased significantly （P<0.05）. Compared with the model group， the expression of IDE increased significantly in the LGZGT-M and LGZGT-H groups （P<0.05）. Compared with the model group， the expression of CTSD increased significantly in the LGZGT-L， LGZGT-M， LGZGT-H， and donepezil hydrochloride groups （P<0.05）. The average fluorescence intensity of CTSB in the model group was significantly lower than that in the normal group （P<0.05）. Compared with the model group， the average fluorescence intensity of CTSD in the LGZGT-L， LGZGT-M， LGZGT-H， and donepezil hydrochloride groups increased significantly （P<0.05）. The intracellular content of Aβ1-42 in cells in the model group was significantly higher than that in the normal group （P<0.05）. After administration， compared with the model group， the intracellular content of Aβ1-42 in cells in the LGZGT-L， LGZGT-M， LGZGT-H， and donepezil hydrochloride groups decreased significantly （P<0.05）， and LGZGT-containing serum reduced Aβ1-42 in a dose-dependent manner （P<0.05）. ConclusionLGZGT has a protective effect on Aβ1-42-induced AS and can promote the degradation of Aβ. Its mechanism may be related to reducing Aβ toxicity， enhancing cell viability， promoting the expression of IDE， CTSD， and CTSB， and restoring lysosomal function.
RESUMEN
ObjectiveTo study the effect of serum containing Sanwubai San on TGF-β1 induced epithelial mesenchymal transition （EMT） of human gastric cancer SGC-7901 cells and its mechanism in vitro based on transforming growth factor-β/Smad（TGF-β/Smad）signaling pathway. MethodTwenty-eight male SD rats （SPF grade， three months） were randomly divided into blank group and Sanwubai low （0.031 25 g·kg-1·d-1， ig）， medium （0.062 5 g·kg-1·d-1， ig） and high （0.125 g·kg-1·d-1， ig） dose groups， seven in each group. The blank group was given the same volume of ultrapure water （ig）. The gavage was performed once a day for seven consecutive days. The serum containing the drug was taken from the abdominal aorta 45 min after the last administration. Cell counting kit-8 （CCK-8） method was used to detect the effect of serum in Sanwubai San high dose group on the activity of SGC-7901 cells. Changes of cell morphology after treatment with TGF-β1 and serum containing Sanwubai San were observed by microscopy， and the migration rate and invasion rate of the SGC-7901 cells were detected by cell scratch assay and transwell assay， respectively. Western blot was used to detect the expression of E-cadherin， snail， TGF-β1， Smad3， p-Smad3 and Smad7 proteins. ResultCompared with the blank group， 10%， 15% and 20% high-dose Sanwubai San inhibited the activity of SGC-7901 cells in a concentration and time dependent manner. Compared with the conditions in the blank group， the cells in the model group lost spindle shape， and most cells became round and long. Compared with the model group， the Sanwubai San groups had decreased pseudopodia and small cells with the morphology returning to normal. Compared with the conditions in the blank group， enhanced ability of cell migration and invasion （P<0.01）， lowered expression of E-cadherin and Smad7 （P<0.01）， and increased expression of Snail， p-Smad3 and TGF-β1 （P<0.01） were found in the model group， with the total protein level of Smad3 remaining unchanged. Compared with the conditions in the model group， the cell migration ability was inhibited in the Sanwubai San high and medium dose groups （P<0.01） after 24 h， and the ability was inhibited in all three Sanwubai San groups after 48 h （P<0.01）， while the invasion ability was enhanced. In addition， the Sanwubai San high and medium dose groups had elevated expression of E-cadherin （P<0.01） and Smad7 （P<0.01）， and decreased expression of Snail （P<0.01）， and the expression of TGF-β1 and p-Smad3 was down-regulated in the three Sanwubai San groups （P<0.01）. ConclusionSanwubai San could inhibit TGF-β1 induced EMT in SGC-7901 cells， and its mechanism might be related to the regulation of TGF-β/Smad signaling pathway.