Effect Of Phytosterols In Pine Pollen On Growth、Differentiation、 Glycometabolism Of3T3-F442A Preadipocyte

Masson pine pollen, also known as “Song Huang” in Chinese traditional medicine,is the dry pollen of masson pine, Chinese pine or the same genera which belongs topinaceae family. It is the traditional pollen variety for food and medicine whichpossessing anti-aging, anti-fatigue, hypoglycemic effect and other pharmacologicaleffects. As a kind of natural components, phytosterols have received wide attentiondue to the prevention of cardiovascular diseases by lowering cholesterol in humanserum.Objective:Through the cultivation of3T3-F442A preadipocytes, we studied the effect ofphytosterols in pollen on growth、differentiation and glycometabolism of3T3-F442Apreadipocyte. By detecting the expression of AMPK、GLUT1content in the matureadipocytes, we tried to study the sterol in pollen affects the glycometabolism in theinternal mechanism of adipocytes.Methods:We detected the effect of phytosterols in pollen on growth of3T3-F442Apreadipocyte in24h,48h and96h. Then we cultured3T3-F442A preadipocytes byinsulin and detected the effect of phytosterols in pollen on differentiation of3T3-F442A preadipocyte by oil red O staining and the triglyceride level kit. In thenext step, we tested the effect of phytosterols in pollen on the growth activity of3T3-F442A adipocyte by MTT assay. In the last step, we detected the effect ofphytosterols in pollen on glycometabolism of3T3-F442A adipocyte by determinationof glucose kit. And then we detected the expression of AMPK、GLUT1content byAMPK、GLUT1ELISA Kit.Results:1、After24h, different concentrations of phytosterols in pollen significantly inhibitedthe growth activity of3T3-F442A preadipocyte(P＜0.01).2、Oil red O staining showed that cells became greatly changed round, a smallamount of lipid droplets in the cytoplasm around the nucleus. After6days, smalllipid droplets increased significantly, start with “Ring Ring”-like structure. In the groups of sterol in pollen, the differentiation rate increased with the increasingconcentration. In the groups of β-sitosterol also promoted the effect ofdifferentiation, which the6.4μg/mL group＇s differentiation rate was the highest.The result of content of triglyceride determination showed that three kinds ofmaterial increased the content of triglyceride, which the3.2μg/mL,6.4μg/mL and12.8μg/mL of sterol in pollen significantly enhanced the content of triglyceride (P＜0.01) with dosage-effect relationship.3、We tested the effect of phytosterols in pollen on the growth activity of3T3-F442Aadipocyte by MTT assay. The result showed that6.4μg/mL of phytosterols inpollen（96h）significantly inhibited the growth activity of3T3-F442A adipocyte(P＜0.01).4、We detected the effect of phytosterols in pollen on glycometabolism of3T3-F442Aadipocyte by determination of glucose kit. We found that except the group of6.4μg/mL of cholesterol, the other groups inhibited the glycometabolism of3T3-F442A adipocyte. And0.8μg/mL of phytosterols in pollen（24h）significantinhibited the glycometabolism of3T3-F442A adipocyte(P＜0.05).5、 Compared with the control, the3.2μg/mL,6.4μg/mL and12.8μg/mL ofphytosterols in pollen significantly inhibited the content of AMPK、GLUT1(P＜0.01).Conclusion:The groups of phytosterols in pollen had inhibition effects on growth of3T3-F442A preadipocyte. But it can promote the differentiation of3T3-F442Apreadipocyte and inhibit the growth activity of3T3-F442A adipocyte、glycometabolism、AMPK and GLUT1content.