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| Apoptosis and autophagy of podocytes studied based on “tonifying kidney collaterals and invigorating spleen and kidney” |
| Hits 1387 Download times 1001 Received:January 02, 2019 |
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| DOI
10.11656/j.issn.1672-1519.2019.07.17 |
| Key Words
diabetic nephropathy;Tongluo Yishen Decoction;podocytes;apoptosis;autophagy;relationship |
| Author Name | Affiliation | E-mail | | FAN Zenghui | Shaanxi University of Chinese Medicine, Xianyang 712046, China | | | MA Fengfeng | Shaanxi University of Chinese Medicine, Xianyang 712046, China | | | LI Xiaohui | Shaanxi University of Chinese Medicine, Xianyang 712046, China | jyhtt20050418@126.com | | CHEN Liming | Shaanxi University of Chinese Medicine, Xianyang 712046, China | | | GU Haorong | Shaanxi University of Chinese Medicine, Xianyang 712046, China | |
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| Abstract
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| [Objective] Based on the treatment method of "tonifying kidney and collaterals,tonifying spleen and kidney",to investigate the regulatory effect of Tongluo Yishen Decoction on the expressions of autophagy and apoptosis proteins SIRT1,BNIP3,P62 and bcl-2 in mouse glomerular podocytes cultured in vitro with high glucose. [Methods] Forty mature SPF-grade SD male rats were randomly divided into the normal group,the large,medium and small dose groups of traditional Chinese medicine,10 rats in each group. According to the method of human and rat body surface area conversion,the concentration of traditional Chinese medicine required by rats was calculated:large dose concentration (4.76 g/mL),medium dose concentration (2.38 g/mL),and small dose concentration (1.19 g/mL). After 10 days of gavage,the drug-containing serum was taken for later use.Podocytes were divided into 6 groups:normal group (5.6 mmol/L glucose+10% normal rat serum),high glucose group (30 mmol/L glucose+10% normal rat serum),Tongluo Yishen Decoction large/medium/small dose intervention groups (30 mmol/L glucose+10% Tongluo Yishen Decoction large/medium/small dose rat serum) and hypertonic group (24.5 mmol/L mannitol+10% normal rat serum). The cells were cultured for 48 h according to the requirements of each group and collected. The apoptosis status and morphological changes of podocytes in each group were observed after fluorescence staining with Hoechst 33342. The apoptosis rate of podocytes was measured by flow cytometry. Expression levels of SIRT1,BNIP3,P62 and Bax proteins in cells were detected by Western Blot. [Results] The results of flow cytometry showed that the medium and small dose of Tongluo Yishen Decoction could reduce the apoptosis rate of podocytes induced by high glucose (P<0.01 or P<0.05) and the high dose group could not improve apoptosis (P>0.05). The observation results of Hoechst33342 fluorescence staining also confirmed that the medium and small dose groups of Tongluo Yishen Decoction could reduce the apoptosis rate of podocytes induced by high glucose. Western blot results showed that compared with the high-glucose group,SIRT1 protein expression was increased in the medium and small dose groups of tongluo yishen prescription,while BNIP3,P62 and Bax protein expression was decreased (P<0.05 or P<0.01). Protein expressions of SIRT1,BNIP3,P62 and Bax were not significantly changed in the high-dose group (P>0.05). [Conclusion] Medium and small doses of Tongluo Yishen Decoction can activate autophagy to reduce apoptosis of podocytes in vitro under the stimulation of high glucose,and reduce apoptosis rate and expression of apoptotic protein. |
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