| Diabetes, of which the main characteristic is disorder of glucolipid absorption, has become one of the diseases damaging human health. And the prevalence of diabetes has also increased year by year. Therefore, the research into how to prevent the disease has become hot.Grape seed procyanidin extract (GSPE) is a kind of complex mixture of polyphenols, the main monomers of GSPE are caatechin, epicatechin and epigallocatechin gallate. These monomers connecting together in many different ways form different degrees of GSPE. We find that the polymers grape seed procyanidins (OPC)’s can adjust glucolipid absorption in T2DM rats, and some monomers can increase the glucose uptake of skeletal muscle cell in vitro.The research into GSPE mainly concentrates on vivo experiments in glucose absorption domain at present, while the vitro experiments at cellular level of the research are not carried on sufficiently. Both the researchs of GSPE are measured as a complex extraction at present,so it is necessary to study the effects of GSPE with different degree in vitro experiments.The purpose of my work is to find out the effects of GSPE with different degree of polymerization on glucose absorption in L6 rat skeletal muscle cells. And we also investigated the method of rapidly inducing the L6 rat skeletal muscle cells differ. The results are as follows:(1) Reducing the concentration of serum can promote the differentiation of L6 rat skeletal muscle cells:the grown well L6 rat skeletal muscle cells were trypsinizated and then cultured in 96 well plates. When the L6 rat skeletal muscle cells grew with adherence, the high glucose DMEM containing 2% fetal bovine serum (FBS) or 2% horse serum (HS) or 10% FBS was added into the L6 rat skeletal muscle cells. Each group repeats 8 time, and the L6 rat skeletal muscle cells were cultured in 37 ℃,5% CO2 incubator. Change the medium after one day and observe the differentiation of L6 rat skeletal muscle cells. We find that the L6 rat skeletal muscle cells cultured in high glucose DMEM containing 10% FBS were differentiated fully after 11 days, but the cell cultured in high glucose DMEM containing 2% FBS or 2%HS were differentiated fully after 6 days.(2) High concentrations of different polymerization degree GSPE affect the differentiation of L6 rat skeletal muscle cells:when different concentrations (0μg/mL、25 μg/mL、50μg/mL、 100μg/mL、 200μg/mL) and different polymerization degrees(oligomeric group mDP=3.3, high polymer group-I mDP=6.4, high polymer group-II mDP=10.5) of GSPE are added into the fully differentiated L6 rat skeletal muscle cells for 12 hours, we find out with increasing concentration, the growth state of the cells gradually deteriorate, cell activity gradually weakened, when the effect of the concentration is 200μg/mL, cells lysis and apoptosis gradually.(3) High concentrations of different polymerization degree GSPE lower the activity of L6 rat skeletal muscle cells: ① The activity of cells decreased with the increase of concentration.When the concentration is higher than 25μg/mL, all the 3 different polymerization degrees of GSPE inhibited the differentiation of L6 rat muscle cell activity, the effect of the inhibition differs significantly with different concentrations (P<0.05). The inhibition rate of L6 rat muscle cell activity by oligomeric group GSPE from low concentration to high concentration is 5.5±3.7%、 30.1±2.6%、 72.3±1.4%、 90.1±0.4%. The inhibition rate of L6 rat muscle cell activity by high polymer group-I from low concentration to high concentration is 18.7±3.3%、 45.0±2.3%、79.7±0.9、90.5±0.4%. The inhibition rate of L6 rat muscle cell activity by high polymer group-II from low concentration to high concentration is 19.7±4.0%、 43.7±2.8%、 80.5±9.7%、91.3±3.5%.②nder the condition of the same concentration, different degree of polymerization GSPE showed different effects on the activity of differentiated L6 rat skeletal muscle cells. When the concentration are 25μg/mLand 50μg/mL,compared with high polymer group-I and high polymer group-II,oligomeric group have effect on the activity of L6 rat skeletal muscle cells,the difference was significant.(P<0.05).The difference is not significant when compared high polymer group-I with high polymer group-Ⅱ.When the concentrition were 100μg/mL and 200μg/mL,the difference is not significant among the three groups.(4) Different degree of polymerization GSPE promote glucose absorption of differentiated L6 rat skeletal muscle cells. The study found when the oligomerization GSPE concentrations are below 20μg/mL, it can significantly promote the differentiation of L6 rat rat muscle cells glucose uptake (P<0.05). The rate of glucose uptake increases by 15.83%±3.20,24.57%±1.41,43.43%±1.97,8.70%±1.54 after added oligomeric group GSPE from low concentration to high concentration. When the high polymer group-I, high polymer group-II are compared with the control group, the difference was not significant.In summary, different concentrations of GSPE have different effects on the differentiation of L6 rat muscle cell growth and glucose absorption. Higher concentration of GSPE inhibit the differentiation of L6 rat muscle cell; Lower concentration of GSPE promote the glucose consumption of L6 rat muscle cell. The effect is related to the polymerization degree, and in a concentration dependent manner. Tests showed that with increasing degree of polymerization, the effect gradually weakened. |
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