Exploration on effects of Modified Zhenwu Decoction on mitochondrial biosynthesis in HK-2 cells based on the oxidative stress and hypoxia
Objective:To observe the effects of Modified Zhenwu Decoction on the expression of ROS,PHD2,HIF-1 α,PGC-1 α,NRF1,NRF2 and TFAM in HK-2 cells treated with hypoxia and LPS,the potential mechanism of Modified Zhenwu Decoction delaying the progression of chronic renal failure(CRF)was investigated.Methods:HK-2 cells were divided into normal group(A),model group(B),blank serum group(C),drug-containing serum group(D),FM19G11 group(E)and NAC group(F).After 72 h of intervention,the ROS levels were detected by flow cytometry.The expression of PGC-1 α was detected by immunofluorescence.The morphological changes of mitochondria were observed by electron microscope.Apoptosis was detected by TUNEL kit.The protein expression levels of HIF-1 α,PHD2,PGC-1 α,NRF1,NRF2 and TFAM were detected by Western Blot.Results:Compared with group A,ROS content in group B significantly increased(P＜0.01),the expression of PGC-1 α significantly decreased(P＜0.01),the protein expressions of PHD2,PGC-1 α,NRF1,NRF2 and TFAM significantly decreased(P＜0.01),while the expressions of HIF-1 α significantly increased(P＜0.01),the number of mitochondria decreased,the inner chamber swelled,and the ridge shifted to the periphery,the number of apoptosis significantly increased(P＜0.0l).Compared with group B,ROS content in cells significantly decreased after advanced intervention with Modified Zhenwu Decoction(P＜0.0 1),the expression of PGC-1 α significantly increased(P＜0.01),the protein expressions of PHD2,PGC-1 α,NRF1,NRF2 and TFAM significantly increased(P＜0.01),while the expressions of HIF-1 α significantly decreased(P＜0.01),the number of mitochondria increased and the pathological changes decreased,the number of apoptosis significantly decreased(P＜0.01).Conclusion:Modified Zhenwu Decoction may reduce cell apoptosis and delay the progression of CRF by inhibiting oxidative stress and hypoxia of proximal curved renal tubular epithelial cells and improving mitochondrial biosynthesis.
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