Effects Of Chongcaoshencha Capsule On Human Mesangial Cells Cultured In Vitro Under High Glucose Condition

Objective:To investigate the effects of Chongcaoshencha capsule on humanmesangial cells cultured in vitro under high glucose condition and explorethe nephroprotective mechanism of Chongcaoshencha capsule for DN.Methods:We first get the rats serum with different concentration of Chongcaoshenchacapsule and fosinopril, and then study the effects of the serum onHMCs. The cultured HMCs were divided into normal control group, highglucose group, High-dose, Middle-dose and Low-dose Chongcaoshenchacapsule group and fosinopril group. The HMCs induced by high glucose wereadopted to study. The normal control group HMCs were cultured with normalrat serum RPMI-1640 culture solution under normal glucose condition, highglucose group HMCs were cultured with normal rat serum high glucoseRPMI-1640 culture solution, the high-dose, middle-dose and low-doseChongcaoshencha group HMCs were incubated with different concentrationof Chongcaoshencha capsule rat serum RPMI-1640 culture solution underhigh glucose condition.the fosinopril group HMCs were incubated withfosinopril rat serum RPMI-1640 culture solution under high glucosecondition. The FCS concentration of every group is 10％. After therespective intervention of 24 hours, 48 hours and 72 hours, the HMCsproliferation was assessed by MTT assay. After culturing 48 and 72 hours byadding different serum respectively, transforming growth factor-betal(TGF-β1) , Fibronectin (FN) , Collagen-Ⅳ(Col-Ⅳ) , laminin (LN)secretion was determined by ELISA. The real time PCR was used to detectthe expression of transforming growth factor beta 1mRNA (TGF-β1mRNA)and plasminogen activator inhibitor-1 mRNA (PAI-1 mRNA)Result:(1) The high-dose Chongcaoshencha capsule serum can inhibit theproliferation of mesangial cells under high glucose condition (30mmol/L) at 24h,48h and 72h (p＜0.05, p＜0.01) , and showed better effect than thefosinopril serum. The middle-dose and low-dose Chongcaosheneha capsulecan inhibit the proliferation of mesangial cells under high glucose condition(30mmol/L) at 48h and 72h (p＜0.05) . (2) TheTGF-β1 secretion of thehigh-dose and middle-dose Chongcaoshencha capsule group is lower thanthat of the high glucose group at 48h and 72h (p＜0.05, p＜0.01) . Thelow-dose Chongcaoshencha capsule and fosinopril can decrease the secretionof TGF-β1 at 72h (p＜0.05) . Chongcaoshencha capsule has better effect. (3)The secretion of Col-Ⅳof Chongcaoshencha capsule group and thefosinopril group is lower than that in the high glucose group at 48h and 72h(p＜0.05, p＜0.01) . (4) The secretion of FN and LN of the high-doseChongcaoshencha capsule group is lower than that in the high glucose groupat 48h and 72h (p＜0.05, p＜0.01 ) . The middle-dose and low-doseChongcaoshencha capsule serum and fosinopril serum can decrease thesecretion of FN,LN at 72h (p＜0.05, p＜0.01) . (5) The expression ofTGF-β1mRNA and PAI-1mRNA of the high-dose and middle-doseChongcaoshencha capsule group and the fosinopril group are lower than thatof the high glucose group, and the expression of high-dose Chongcaoshenehacapsule group is the lowest (p＜0.05)Conclusion:Chongcaoshencha capsule not only can inhibit high glucose-inducedproliferation of HMCs, but also can decrease the secretion of TGF-β1, FN,Col-Ⅳ, LN, and degrade the expression of TGF-β1mRNA and PAI-1mRNA inthe high glucose cultured condition, so to delay the progression of DN.