Objective To explore the influence of Linggui Zhugan Decoction(LGZGD) on high glucose induced podocyte autophagy.Methods LGZGD containing serum was prepared by intragastric administation of 4.2 g/kg(low dose), 8.4 g/k...Objective To explore the influence of Linggui Zhugan Decoction(LGZGD) on high glucose induced podocyte autophagy.Methods LGZGD containing serum was prepared by intragastric administation of 4.2 g/kg(low dose), 8.4 g/kg(medium dose), and 12.6 g/kg(high dose) LGZGD into SD rats respectively. MPC5 and AB8/13 podocyte cells were treated with 60 mmol/L glucose to establish diabetic nephropathy podocyte model in vitro. Both podocytes were divided into control group, high glucose group, low dose LGZGD group, medium dose LGZGD group, and high dose LGZGD group, respectively. For the three LGZGD groups, before LGZGD intervention, podocytes were treated with 60 mmol/L glucose for 3 days. After treated with LGZGD containing serum, cells were collected to analyze cell migration using Transwell assay, proliferation using CCK8, apoptosis and cell cycle using flow cytometry, autophagosome formation using transmission electron microscopy, and expression levels of Beclin-1, Atg5, LC3II/I, and P62 proteins using Western blot.Results Compared with the control group, the proliferation and migration of MPC5 and AB8/13 cells in the high glucose group slightly decreased, whereas these parameters restored after intervention with low and medium concentrations of LGZGD, with the medium dose LGZGD having the better effect(P < 0.05). Flow cytometry showed that the medium dose LGZGD group had a significantly lower apoptosis rate(P < 0.05) and higher survival rate(P > 0.05) compared to the high dose LGZGD group. High glucose arrested podocytes in G1 phase, whereas LGZGD shifted podocytes from being predominant in G1 phase to G2 phase. High dose LGZGD significanly reduced high glucose-increased autophagosome formation in both podocytes(P < 0.05). Western blot analysis showed that Beclin-1, Atg5, LC3II/I, and P62 expressions were increased in MPC5 cells treated with high glucose and reversed after adminstration of low and medium doses of LGZGD(P < 0.05).Conclusion LGZGD reduced apoptosis and enhanced autophagy in high glucose treated podocytes via regulating Beclin-1/LC3II/I/Atg5 expression.展开更多
Objective： To explore the effects of alternate-day-fasting （ADF） therapy combined with Lingguizhugan Decoction （LD）on blood lipid profiles of hyperlipidemic rats. Methods： Rats were randomly assigned into high-f...Objective： To explore the effects of alternate-day-fasting （ADF） therapy combined with Lingguizhugan Decoction （LD）on blood lipid profiles of hyperlipidemic rats. Methods： Rats were randomly assigned into high-fat-diet （HF） group andnormal-diet （ND） group. Hyperlipidemic rats fed with high-fat-diet for 5 weeks were randomly divided into ADF group,alternate-day-fasting with LD （ALG） group and model control （MC） group. The rats in ALG and ADF group weredeprived of food for 24 h every other day for 4 weeks. Rats in ALG group were administrated with LD at fasting day.After 4 weeks of ADF therapy, plasma TC, TG, LDL-c and HDL-c were measured in each group. Expression of miR-143and PPAR-γ protein from adipose was also analyzed. Results： When compared with MC group, after 4 weeks of ADF orcombined ADF and LD therapy, the body weight was evidently reduced in ADF and ALG groups （P = 0.028, P = 0.036by wk 8）. The levels of plasma TC and TG decreased in ADF group and ALG group, which were significantly lower thanthose in MC group （P 〈 0.001, P = 0.045; P 〈 0.001, P = 0.005）. However, the body weight and level of TC and TG inALG group showed non-statistical difference in comparison with ADF group （ALG vs. ADF, P 〉 0.05）. Expression ofmiR-143 and PPAR-γ were higher in MC group than that in NC group （P 〈 0.001）. Compared with MC group,expression of miR-143 and PPAR-γ were significantly decreased in ADF （P = 0.038, P = 0.015） and ALG （P = 0.007, P〈 0.001） groups. When compared with ADF group, expression of miR-143 and PPAR-γ were significantly decreased inALG （P = 0.041, P = 0.046） group. Conclusion： ADF therapy alone not only reduced blood lipids, but also inhibitedmiR-143 and PPAR-γ protein expression in visceral adipose tissue. However, LD couldn’t reduce the levels of bloodlipid profiles more effectively than using ADF alone. Perhaps the effects of LD combined with ADF in the prevention ofhyperlipidemia need further exploration.展开更多
基金supported by Guangdong Bureau of Traditional Chinese Medicine (20211082)
文摘Objective To explore the influence of Linggui Zhugan Decoction(LGZGD) on high glucose induced podocyte autophagy.Methods LGZGD containing serum was prepared by intragastric administation of 4.2 g/kg(low dose), 8.4 g/kg(medium dose), and 12.6 g/kg(high dose) LGZGD into SD rats respectively. MPC5 and AB8/13 podocyte cells were treated with 60 mmol/L glucose to establish diabetic nephropathy podocyte model in vitro. Both podocytes were divided into control group, high glucose group, low dose LGZGD group, medium dose LGZGD group, and high dose LGZGD group, respectively. For the three LGZGD groups, before LGZGD intervention, podocytes were treated with 60 mmol/L glucose for 3 days. After treated with LGZGD containing serum, cells were collected to analyze cell migration using Transwell assay, proliferation using CCK8, apoptosis and cell cycle using flow cytometry, autophagosome formation using transmission electron microscopy, and expression levels of Beclin-1, Atg5, LC3II/I, and P62 proteins using Western blot.Results Compared with the control group, the proliferation and migration of MPC5 and AB8/13 cells in the high glucose group slightly decreased, whereas these parameters restored after intervention with low and medium concentrations of LGZGD, with the medium dose LGZGD having the better effect(P < 0.05). Flow cytometry showed that the medium dose LGZGD group had a significantly lower apoptosis rate(P < 0.05) and higher survival rate(P > 0.05) compared to the high dose LGZGD group. High glucose arrested podocytes in G1 phase, whereas LGZGD shifted podocytes from being predominant in G1 phase to G2 phase. High dose LGZGD significanly reduced high glucose-increased autophagosome formation in both podocytes(P < 0.05). Western blot analysis showed that Beclin-1, Atg5, LC3II/I, and P62 expressions were increased in MPC5 cells treated with high glucose and reversed after adminstration of low and medium doses of LGZGD(P < 0.05).Conclusion LGZGD reduced apoptosis and enhanced autophagy in high glucose treated podocytes via regulating Beclin-1/LC3II/I/Atg5 expression.
文摘Objective： To explore the effects of alternate-day-fasting （ADF） therapy combined with Lingguizhugan Decoction （LD）on blood lipid profiles of hyperlipidemic rats. Methods： Rats were randomly assigned into high-fat-diet （HF） group andnormal-diet （ND） group. Hyperlipidemic rats fed with high-fat-diet for 5 weeks were randomly divided into ADF group,alternate-day-fasting with LD （ALG） group and model control （MC） group. The rats in ALG and ADF group weredeprived of food for 24 h every other day for 4 weeks. Rats in ALG group were administrated with LD at fasting day.After 4 weeks of ADF therapy, plasma TC, TG, LDL-c and HDL-c were measured in each group. Expression of miR-143and PPAR-γ protein from adipose was also analyzed. Results： When compared with MC group, after 4 weeks of ADF orcombined ADF and LD therapy, the body weight was evidently reduced in ADF and ALG groups （P = 0.028, P = 0.036by wk 8）. The levels of plasma TC and TG decreased in ADF group and ALG group, which were significantly lower thanthose in MC group （P 〈 0.001, P = 0.045; P 〈 0.001, P = 0.005）. However, the body weight and level of TC and TG inALG group showed non-statistical difference in comparison with ADF group （ALG vs. ADF, P 〉 0.05）. Expression ofmiR-143 and PPAR-γ were higher in MC group than that in NC group （P 〈 0.001）. Compared with MC group,expression of miR-143 and PPAR-γ were significantly decreased in ADF （P = 0.038, P = 0.015） and ALG （P = 0.007, P〈 0.001） groups. When compared with ADF group, expression of miR-143 and PPAR-γ were significantly decreased inALG （P = 0.041, P = 0.046） group. Conclusion： ADF therapy alone not only reduced blood lipids, but also inhibitedmiR-143 and PPAR-γ protein expression in visceral adipose tissue. However, LD couldn’t reduce the levels of bloodlipid profiles more effectively than using ADF alone. Perhaps the effects of LD combined with ADF in the prevention ofhyperlipidemia need further exploration.
