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 therapeutic effect of Linggui Zhugan Decoction on nonalcoholic fatty liver disease through in vivo animal experiments, and to explore the mechanism of action of Linggui Zhugan Decoction in th...Objective: To explore the therapeutic effect of Linggui Zhugan Decoction on nonalcoholic fatty liver disease through in vivo animal experiments, and to explore the mechanism of action of Linggui Zhugan Decoction in the intervention of nonalcoholic fatty liver disease through network pharmacology and molecular docking technology. Methods: A rat model of non- alcoholic fatty liver disease was established after 20 weeks of high-fat feeding, and the rats were divided into six groups, normal group, model group, simvastatin group (4 mg/kg), low- dose group of Linggui Zhugan Decoction (2.1 g/kg), medium-dose group of Linggui Zhugan Decoction (4.2 g/kg), and high-dose group of Linggui Zhugan Decoction (8.4 g/kg), and the liver morphological changes of HE staining and oil red O staining after the intervention of Linggui Zhugan Decoction were observed, and the microplate reader detected aspartate aminotransferase, alanine aminotransferase, Four levels of blood lipids. Network pharmacology combined with molecular docking was used to screen the potential mechanism and target of Linggui Zhugan Decoction for the intervention of non-alcoholic fatty liver disease. Results: Linggui Zhugan Decoction significantly improved the levels of alanine aminotransferase and alanine aminotransferase in model rats (P<0.05). HE staining and oil red O staining showed that Linggui Zhugan Decoction significantly reduced liver lipid deposition in rats in the model group. Linggui Zhugan Decoction could significantly reduce the quadruplical levels of blood lipids in model rats (P<0.05). Network pharmacology screened out 10 key targets and 5 key signaling pathways;The molecular docking results showed that the first 20 active ingredients all had good binding ability to the top 10 targets. Conclusion: The results show that Linggui Zhugan Decoction has the effect of regulating blood lipids and improving liver tissue lesions in non-alcoholic fatty liver disease, and preliminarily verifies the regulatory effect of Linggui Zhugan Decoction on lipid metabolism-related genes.展开更多
Objective To observe the clinical effect of fire needle combined with Linggui Bafa for post-herpetic neuralgia （PHN）. Methods One hundred and twenty patients were randomly divided into an observation group and a con...Objective To observe the clinical effect of fire needle combined with Linggui Bafa for post-herpetic neuralgia （PHN）. Methods One hundred and twenty patients were randomly divided into an observation group and a control group with 60 cases each. For observation group, fire needle was applied to prick Zusanli （足三里 ST 36）, Yanglingquan （阳陵泉 GB 34）, Taichong （太冲 LR 3）, Sanyinjiao （三阴交 SP 6） and partial Ashi points around the lesion, and Linggui Bafa was combined to perform acupuncture therapy, once every other day, 10 times as a course of treatment; for the control group, only fire needle was applied. After 2 courses of treatment, the comparison of the value of visual analogue scale （VAS） and the total effective rate for the two groups were carried out. Results The VAS value of the observation group after treatment was obviously lower than that before treatment （2.28±2.08 vs 6.12±1.96） and that of the control group （2.28±2.08 vs 3.62±2.90）, there were significant differences （all P〈0.01）. The total effective rate of the observation group was obviously higher than that of the control group [93.3% （56/60） vs 76.7% （46/60）], the difference between the two groups was statistically significant （P〈0.05）. Conclusion The effect of fire needle combined with Linggui Ba.fa for PHN is obviously superior to that of single fire needle therapy.展开更多
Objective To evaluate the therapeutic effect and safety of time-oriented points opening of Linggui Bafa for Bell＇s palsy and provide a new idea for the clinical treatment of this problem. Methods Fifty-four cases inc...Objective To evaluate the therapeutic effect and safety of time-oriented points opening of Linggui Bafa for Bell＇s palsy and provide a new idea for the clinical treatment of this problem. Methods Fifty-four cases included in the study were randomized into two groups, and the treatment group was treated with the routine acupuncture combined with time-oriented points opening of Linggui Bafa. The control group was treated with the routine acupuncture only, Hegu （合谷 El 4）, Kunlun （昆仑 BL 60）, Jiache （颊车 ST 6）, Dicang （地仓 ST 4）, Xiaguan （下关 ST 7）, Cuanzhu （攒竹 BL 2）, Yangbai （附白 GB 14）, and Quanliao （颧髎 SI 18） were selected as the main points. The manipulation and time of retaining needles were as same as the treatment group. Both groups were treated once a day, 5 days made one session and 4 sessions were required in all. Modified Portmann＇s scale, Horse-Brackmann grading scale （H-B）, FDIP and FDIS were used to assess the therapeutic effect after the treatment. Results The curative and improvement rate in the treatment group was 80.8% （22/26）, and that in the control group was 66.7% （16/24）, with the statistically significant difference （P〈O.05）, the comparison of scores of Portmann＇s scale, H-B, FDIP and FDIS in the same group and between the two groups before and after the treatment also had statistically significant differences （all P〈0.05）. Conclusion The method of time-oriented points opening of Linggui Bafa for Bell＇s palsy is better than the routine acupuncture for Bell＇s palsy.展开更多
Objective:To investigate the inhibitory effect of Linggui Zhugan Decoction(LZD,苓桂术甘汤)on the ventricular remodeling(VR)after acute myocardial infarction(AMI)and related mRNA and proteins expression in transforming...Objective:To investigate the inhibitory effect of Linggui Zhugan Decoction(LZD,苓桂术甘汤)on the ventricular remodeling(VR)after acute myocardial infarction(AMI)and related mRNA and proteins expression in transforming growth factor-beta 1(TGF-β1)/Smad signaling pathway,and explain its putative mechanism.Methods:A VR model was generated by ligation of coronary artery in mice.Two weeks after surgery,60 mice were randomly divided into the model group,the sham-operation group(distilled water),the positive control group(2.4 mg/kg simvastatin),and the low-,medium-and high-dose LZD groups(2.1,4.2,8.4 g crude drug/kg,respectively)by a random number table,10 mice in each group.Mice in each group was treated for 4 weeks.Changes of hemodynamics indices and cardiac weight index were detected by the PowerLab data acquisition and analysis recording instrument.Morphology changes of myocardial tissue were observed by hematoxylin-eosin and Masson staining.The expressions of TGF-β1,Smad2,Smad3,p-Smad2 and p-Smad3 in myocardial tissue were detected by Western blotting.The mRNA expressions of TGF-β1,Smad2 and Smad3 were detected by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).The expressions of matrix metalloprotein 2(MMP2),MMP9,collagenⅠand collagen Ⅲ were observed by immunohistochemical methods.Results:VR mice showed significant dysfunction in hemodynamic indices and cardiac structure and function.Compared with the shamoperation group,myocardial tissue damage,interstitial fibrosis occurred in the model mice,left ventricular systolic pressure(LVSP),left ventricular pressure maximum contraction rate(+dp/dtmax)and left ventricular pressure maximum relaxation rate(-dp/dtmax)decreased significantly(all P<0.01),while left ventricular end-diastolic pressure(LVEDP),cardiac weight index and left ventricular weight index elevated significantly,meanwhile TGF-β1,p-Smad2,p-Smad3,Smad2,Smad3,MMP2,MMP9,collagen Ⅰ,collagen Ⅲ protein expressions in myocardial tissue and TGF-β1 Smad2 and Smad3 mRNA expressions increased significantly(all P<0.01).Compared with the model group,LZD could significantly improve the pathological changes of myocardial tissue,increase LVSP,+dp/dtmax and-dp/dtmaxf lower LVEDP,reduce the whole heart weight index and left ventricular weight index and inhibit the over-expressions of TGF-β1f p-Smad2,p-Smad3,Smad2,Smad3,MMP2,MMP9,collagenⅠand collagenⅢproteins in myocardial tissue and mRNA expressions of TGF-β1 Smad2 and Smad3(P<0.05 or P<0.01).Conclusion:LZD can significantly suppress VR induced by AMI,and its underlying mechanism may be associated with its inhibitory effect on the TGF-β1/Smad signaling pathway.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.
基金Anhui Natural Science Foundat ion General Project(No.2208085MH277)。
文摘Objective: To explore the therapeutic effect of Linggui Zhugan Decoction on nonalcoholic fatty liver disease through in vivo animal experiments, and to explore the mechanism of action of Linggui Zhugan Decoction in the intervention of nonalcoholic fatty liver disease through network pharmacology and molecular docking technology. Methods: A rat model of non- alcoholic fatty liver disease was established after 20 weeks of high-fat feeding, and the rats were divided into six groups, normal group, model group, simvastatin group (4 mg/kg), low- dose group of Linggui Zhugan Decoction (2.1 g/kg), medium-dose group of Linggui Zhugan Decoction (4.2 g/kg), and high-dose group of Linggui Zhugan Decoction (8.4 g/kg), and the liver morphological changes of HE staining and oil red O staining after the intervention of Linggui Zhugan Decoction were observed, and the microplate reader detected aspartate aminotransferase, alanine aminotransferase, Four levels of blood lipids. Network pharmacology combined with molecular docking was used to screen the potential mechanism and target of Linggui Zhugan Decoction for the intervention of non-alcoholic fatty liver disease. Results: Linggui Zhugan Decoction significantly improved the levels of alanine aminotransferase and alanine aminotransferase in model rats (P<0.05). HE staining and oil red O staining showed that Linggui Zhugan Decoction significantly reduced liver lipid deposition in rats in the model group. Linggui Zhugan Decoction could significantly reduce the quadruplical levels of blood lipids in model rats (P<0.05). Network pharmacology screened out 10 key targets and 5 key signaling pathways;The molecular docking results showed that the first 20 active ingredients all had good binding ability to the top 10 targets. Conclusion: The results show that Linggui Zhugan Decoction has the effect of regulating blood lipids and improving liver tissue lesions in non-alcoholic fatty liver disease, and preliminarily verifies the regulatory effect of Linggui Zhugan Decoction on lipid metabolism-related genes.
文摘Objective To observe the clinical effect of fire needle combined with Linggui Bafa for post-herpetic neuralgia （PHN）. Methods One hundred and twenty patients were randomly divided into an observation group and a control group with 60 cases each. For observation group, fire needle was applied to prick Zusanli （足三里 ST 36）, Yanglingquan （阳陵泉 GB 34）, Taichong （太冲 LR 3）, Sanyinjiao （三阴交 SP 6） and partial Ashi points around the lesion, and Linggui Bafa was combined to perform acupuncture therapy, once every other day, 10 times as a course of treatment; for the control group, only fire needle was applied. After 2 courses of treatment, the comparison of the value of visual analogue scale （VAS） and the total effective rate for the two groups were carried out. Results The VAS value of the observation group after treatment was obviously lower than that before treatment （2.28±2.08 vs 6.12±1.96） and that of the control group （2.28±2.08 vs 3.62±2.90）, there were significant differences （all P〈0.01）. The total effective rate of the observation group was obviously higher than that of the control group [93.3% （56/60） vs 76.7% （46/60）], the difference between the two groups was statistically significant （P〈0.05）. Conclusion The effect of fire needle combined with Linggui Ba.fa for PHN is obviously superior to that of single fire needle therapy.
文摘Objective To evaluate the therapeutic effect and safety of time-oriented points opening of Linggui Bafa for Bell＇s palsy and provide a new idea for the clinical treatment of this problem. Methods Fifty-four cases included in the study were randomized into two groups, and the treatment group was treated with the routine acupuncture combined with time-oriented points opening of Linggui Bafa. The control group was treated with the routine acupuncture only, Hegu （合谷 El 4）, Kunlun （昆仑 BL 60）, Jiache （颊车 ST 6）, Dicang （地仓 ST 4）, Xiaguan （下关 ST 7）, Cuanzhu （攒竹 BL 2）, Yangbai （附白 GB 14）, and Quanliao （颧髎 SI 18） were selected as the main points. The manipulation and time of retaining needles were as same as the treatment group. Both groups were treated once a day, 5 days made one session and 4 sessions were required in all. Modified Portmann＇s scale, Horse-Brackmann grading scale （H-B）, FDIP and FDIS were used to assess the therapeutic effect after the treatment. Results The curative and improvement rate in the treatment group was 80.8% （22/26）, and that in the control group was 66.7% （16/24）, with the statistically significant difference （P〈O.05）, the comparison of scores of Portmann＇s scale, H-B, FDIP and FDIS in the same group and between the two groups before and after the treatment also had statistically significant differences （all P〈0.05）. Conclusion The method of time-oriented points opening of Linggui Bafa for Bell＇s palsy is better than the routine acupuncture for Bell＇s palsy.
基金Supported by the National Natural Science Foundatio of China(No.30973707,81373533,81202631)the Natural Science Foundation of Anhui Province(NO.1508085QH192,1608085QH222)。
文摘Objective:To investigate the inhibitory effect of Linggui Zhugan Decoction(LZD,苓桂术甘汤)on the ventricular remodeling(VR)after acute myocardial infarction(AMI)and related mRNA and proteins expression in transforming growth factor-beta 1(TGF-β1)/Smad signaling pathway,and explain its putative mechanism.Methods:A VR model was generated by ligation of coronary artery in mice.Two weeks after surgery,60 mice were randomly divided into the model group,the sham-operation group(distilled water),the positive control group(2.4 mg/kg simvastatin),and the low-,medium-and high-dose LZD groups(2.1,4.2,8.4 g crude drug/kg,respectively)by a random number table,10 mice in each group.Mice in each group was treated for 4 weeks.Changes of hemodynamics indices and cardiac weight index were detected by the PowerLab data acquisition and analysis recording instrument.Morphology changes of myocardial tissue were observed by hematoxylin-eosin and Masson staining.The expressions of TGF-β1,Smad2,Smad3,p-Smad2 and p-Smad3 in myocardial tissue were detected by Western blotting.The mRNA expressions of TGF-β1,Smad2 and Smad3 were detected by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).The expressions of matrix metalloprotein 2(MMP2),MMP9,collagenⅠand collagen Ⅲ were observed by immunohistochemical methods.Results:VR mice showed significant dysfunction in hemodynamic indices and cardiac structure and function.Compared with the shamoperation group,myocardial tissue damage,interstitial fibrosis occurred in the model mice,left ventricular systolic pressure(LVSP),left ventricular pressure maximum contraction rate(+dp/dtmax)and left ventricular pressure maximum relaxation rate(-dp/dtmax)decreased significantly(all P<0.01),while left ventricular end-diastolic pressure(LVEDP),cardiac weight index and left ventricular weight index elevated significantly,meanwhile TGF-β1,p-Smad2,p-Smad3,Smad2,Smad3,MMP2,MMP9,collagen Ⅰ,collagen Ⅲ protein expressions in myocardial tissue and TGF-β1 Smad2 and Smad3 mRNA expressions increased significantly(all P<0.01).Compared with the model group,LZD could significantly improve the pathological changes of myocardial tissue,increase LVSP,+dp/dtmax and-dp/dtmaxf lower LVEDP,reduce the whole heart weight index and left ventricular weight index and inhibit the over-expressions of TGF-β1f p-Smad2,p-Smad3,Smad2,Smad3,MMP2,MMP9,collagenⅠand collagenⅢproteins in myocardial tissue and mRNA expressions of TGF-β1 Smad2 and Smad3(P<0.05 or P<0.01).Conclusion:LZD can significantly suppress VR induced by AMI,and its underlying mechanism may be associated with its inhibitory effect on the TGF-β1/Smad signaling pathway.
