The Protective Effects And Mechanism Of Rhizoma Nelumbinis On Podocytes Induced By High Glucose

Objective:To observe the proliferation effect of Rhizoma Nelumbinis medicated serum on podocytes in high glucose environment, and the expression of nephrin and podocin in podocytes, so as to explore the protective mechanism that Rhizoma Nelumbinis on podocytes.Methods:Conditionally immortalized mouse podocytes, 37 ℃ incubator with no γ-IFN in10% FBS RPMI medium culture for 14 days, the podocyte differentiation and maturation were randomly divided into seven groups,(1) normal glucose group(NG: 5.5mmol/LGlu),(2) high glucose group(HG:30 mmol/LGlu),(3) hypertonic group(MA:NG+ 24.5 mmol/L mannitol),(4) low-dose Rhizoma Nelumbinis group(RL: HG+Rhizoma 1.5 g/Kg/d medicated serum),(5) medium-dose Rhizoma Nelumbinis group(RM: HG+ Rhizoma 3.0 g/kg/d medicated serum),(6) high-dose Rhizoma Nelumbinis group(RH,HG+ Rhizoma 6.0 g/kg/d medicated serum);(7) Benazepril group(BP,HG+Benazepril medicated serum). The culture fluid of all the above groups contained 10% rat serum. CCK-8 method was used to detect the proliferation of podocytes in 12 h, 24 h, 36 h, 48 h; Changes of nephrin and podocin protein expression in the cells of all the groups after 48 h were detected by Western Blot.Results:1. Effect of different doses Rhizoma Nelumbinis on the proliferation of podocytesinduced by high glucose: Compared with NG group, HG group when cultured 12 h no significant difference in the OD, OD value decreased at 24 h,but there was no statistical significance(P>0.05), 36 h OD value lower was statistically significant(P<0.05), 48 h OD value decreased more significantly(P<0.05), Differences of OD value between MA group at each time point and NG group were not statistically significant(P>0.05); It suggested that the cell proliferation was inhibited after cultured 36 h in high glucose medium, and the inhibition effect was also obvious in 48 h, and the effect was independent of the high osmotic pressure caused by glucose. Podocytes were treated with different doses of Rhizoma Nelumbinis, compared with HG group, the OD values of RH group was increased after cultured 36 h(P<0.05), the OD values of RM, RH and BP group was increased in 48 h(P<0.05); Over the same period the RM, RH group was no significant difference in BP group; It was suggested that middle and high doses of Rhizoma Nelumbinis can antagonize the proliferation inhibition of high glucose on podocyte, and the best time was 48 h.2 Expression of nephrin and podocin protein in the cells of each group: Compared with the NG group, the expression of nephrin and podocin protein in HG group was significantly decreased(P<0.05), MA group had no difference between the expression of two proteins. Compared with HG group, the expression of nephrin and podocin protein in the cells which were intervented by different dose of Rhizoma Nelumbinis and Benazepril were increased. The RM group of podocin protein expression increased significantly(P<0.05), the RH and BP group of nephrin and podocin protein expression increased more significantly(P<0.05).Conclusions:1. High glucose can significantly inhibit the proliferation of podocytes, middle and high dose of Rhizoma Nelumbinis medicated serum can antagonize the proliferation inhibition of high glucose on podocyte.2. Expression of nephrin and podocin protein in high glucose environment wasdecreased, middle and high dose of Rhizoma Nelumbinis group can be raised the two protein expression on podocytes under high glucose environment. It was suggested that Rhizoma Nelumbinis may upregulate in podocytes of nephrin and podocin protein expression, to protect podocytes.