Mechanistic study of lipid metabolism disorders in diabetic kidney disease treated with GLQMP based on network pharmacology,molecular docking and in vitro experiments

Background:In this study,we used network pharmacology and molecular docking combined with vitro experiments to explore the potential mechanism of action of Gualou Qumai pill(GLQMP)against DKD.Methods:We screened effective compounds and drug targets using Chinese medicine systemic pharmacology database and analysis platform and Chinese medicine molecular mechanism bioinformatics analysis tools;and searched for DKD targets using human online Mendelian genetics and gene cards.The potential targets of GLQMP for DKD were obtained through the intersection of drug targets and disease targets.Cytoscape software was applied to build herbal medicine-active compound-target-disease networks and analyze them;protein-protein interaction networks were analyzed using the STRING database platform;gene ontology and Kyoto Encyclopedia of Genes and Genomes were used for gene ontology and gene and genome encyclopedia to enrich potential targets using the DAVID database;and the AutoDock Vina 1.1.2 software for molecular docking of key targets with corresponding key components.In vitro experiments were validated by CCK8,oil red O staining,TC,TG,RT-qPCR,and Western blot.Results:Through network pharmacology analysis,a total of 99 potential therapeutic targets of GLQMP for DKD and the corresponding 38 active compounds were obtained,and 5 core compounds were identified.By constructing the protein-protein interaction network and performing network topology analysis,we found that PPARA and PPARG were the key targets,and then we molecularly docked these two key targets with the 38 active compounds,especially the 5 core compounds,and found that PPARA and PPARG had good binding ability with a variety of compounds.In vitro experiments showed that GLQMP was able to ameliorate HK-2 cell injury under high glucose stress,improve cell viability,reduce TC and TG levels as well as decrease the accumulation of lipid droplets,and RT-qPCR and Western blot confirmed that GLQMP was able to promote the expression levels of PPARA and PPARG.Conclusion:Overall,this study revealed the active compounds,important targets and possible mechanisms of GLQMP treatment for DKD,and conducted preliminary verification experiments on its correctness,provided novel insights into the treatment of DKD by GLQMP.

Based on the theory of“Kidney-blood-bone”to explore the mechanism of GH/IGF-1 system in the prevention and treatment of osteoporosis in Qing'e pill

Objective:To observe the changes of growth hormone(GH)/insulin-like growth factor-1(Igf-1)system in blood,kidney and bone tissue of rats,and to explore the preventive and therapeutic effect of Qing'e pill on osteoporosis due to kidney deficiency and its mechanism,to provide the basis for studying the etiology and pathogenesis of osteoporosis due to kidney deficiency.Methods:The rat osteoporosis model was established by ovariectomy,they were randomly divided into normal group,model group,Western Medicine Group,Jianpi Group,high-dose group,middle-dose group and low-dose group,the levels of serum alkaline phosphatase(ALP),tartrate acid phosphatase(TRACP),growth hormone(GH)and GHR(Ghr)in kidney,bone and blood were determined by Elisa,the contents of insulin-like growth factor-1(Igf-1)and insulin-like growth factor-1 receptor(IGF-1R)were analyzed by SPSS 25.0 software.Results:1.Compared with the normal group,the contents of TRACP and ALP in serum of the model group were significantly increased,suggesting that the rats developed highconversion osteoporosis.2.The levels of serum,bone and renal GHR,Igf-1 and insulin-like growth factor-1 receptor(IGF-1R)in the model group were significantly lower than those in the control group.3.Compared with the model group,the contents of TRACP and ALP in the high,middle and low dose groups were significantly decreased.4.Compared with the model group,the contents of GHR,Igf-1 and IGF-1R in blood,bone and kidney tissue of the high,middle and low dose groups were increased.Conclusion:1.Qing'e pill can improve the bone metabolism of osteoporosis.2.Qinge pill may prevent and cure osteoporosis by increasing the contents of GHR,Igf-1 and IGF-1R in blood,bone and kidney.

Pattern of disharmony between the heart and kidney: Theoretical basis, identification and treatment

'Harmony between the heart and kidney' refers to the physiological relationship between these two zang organs in Chinese medicine,while 'disharmony between the heart and kidney' (also called disharmony between fire and water) refers to the pathological state.The pattern of disharmony between the heart and kidney is widely observed in patients with insomnia,anxiety disorder and menopausal syndrome,etc..In order to gain a full and systematical understanding of this pattern,from the perspective of ancient Chinese philosophy and zang-fu theory in Chinese medicine,we systematically reviewed and discussed the functions and physiological characteristics of the heart and kidney,the origin and development of theories relating to heart-kidney relationship,the pathogenesis and identification of the pattern,as well as the commonly used classical formulas for its treatment,including Coptis and DonkeyHide Gelatin Decoction (Huánglián (E)jiāo Tāng) and Grand Communication Pill (Jiāotài Wán).Two examples of clinical modifications of these formulas in the treatment of insomnia and menopausal syndrome are provided in this article.It should be noted that in clinical practice,these formulas should be used flexibly,and modified in accordance to the condition of the patient.

Liuwei Dihuang Pill(六味地黄丸)Treats Postmenopausal Osteoporosis with Shen(Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression

Objectives： To investigate the mechanism of Liuwei Dihuang Pill （六味地黄丸, LDP） in treating postmenopausal osteoporosis （PMOP） with Shen （Kidney） yin deficiency. Methods： In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group （Group A）, PMOP Shen-yang deficiency group （Group B）, PMOP without Shen deficiency group （Group C）, and control group （Group N）. Real-time polymerase chain reaction （RT-PCR） and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 （CLCF1）, ankyrin repeat and SOCS box containing 1 （ASB1）, and proldneticin 2 （PROK2） genes and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. Results： The mRNA （P〈0.05） and protein （P〈0.01） expression levels of the CLCF1 gone in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gone were obviously up-regulated （P〈0.01）. After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated （both P〈0.01）, and the average bone density of the top femur had significantly increased （P〈0.05）. In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3. Conclusions： The CLCF1 gone is an important gone associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gone expression and activation of the JAK/STAT signaling pathway.

Effect of Zhuchun Pill on Immunity and Endocrine Function of Senile Kidney-Yang Daficiency

Objective: To evaluate the effect of Zhuchun pill (ZCP) on senile Kidney-Yang Deficiency (KYD). Methods: By single blind method, 45 elderly males with KYD were divided randomly into the ZCP group (treated by ZCP) and the control group (treated by placebo, starch capsule) and treated for 3 months.Results: ZCP could greatly improve the main symptoms of KYD, P < 0. 01 in comparison with the control group. After treatment, in the ZCP group, the 3H-TdR lymphocyte transformation rate, levels of serum C3,IgA, IgD, IgG and IgM, superoxide dismutase, plasma adrenocortical hormone, testosterone and cyclic adenosine monophosphate all increased, P < 0. 01, and plasma lipid peroxide decreased, P < 0. 01. While in the control group, no remarkable change was found in these parameters. Conclusion: ZCP could improve the KYD through regulating the immunity, endocrine function and scavenging the free radicals in organism.

Effect of Shexiang Baoxin Pill(麝香保心丸) in Alleviating Early Hypertensive Renal Injury in Rats

Objective:To investigate the effect of Shexiang Baoxin Pill(麝香保心丸,SBP)on early hypertensive renal injury in rats and to explore the possible mechanism.Methods:Twelve-week-old spontaneous hypertensive rats(SHRs)with high-salt diet(dietary containing 8%NaCl)were randomized into the SBP group[40 mg/(kg·d)],losartan potassium group[20 mg/(kg·d)]and saline group by stratified random sampling method,12 in each group.Blood pressure and urea albumin creatinine ratio were measured.After 10 weeks,the expression levels of serum creatinine(Scr),hypersensitive C-reactive protein(hs-CRP),interleukin(IL)-1β,IL-6,tumor necrosis factorα(TNF-α),and transforming growth factorβ(TGF-β)in serum were assessed.Kidney pathology periodate-schiff staining was performed.Semi-quantitative count of macrophage infiltration was determined by immunochemistry of CD68 staining.Real-time quantitative polymerase chain reaction and Western blot were performed to examine the mRNA and protein expressions of Toll-like receptor 4(TLR4),nuclear factorκB(NF-κB),monocyte chemokine peptide(MCP-1),inducible nitric oxide synthase(iNOS),and arginase-1(Arg-1).Results:SBP did not affect the mortality of SHR(P<0.05).SBP significantly reduced the level of elevated blood pressure of SHRs,but the effect was less significantly than that of losartan potassium.SBP decreased urine protein(P<0.01)and the expression levels of IL-1β,IL-6,TNF-α,and TGF-βin serum.The 22-week-old SHRs showed mild proliferation of glomerular endothelial cells,glomerular ischemic lesions,inflammatory cell infiltration in renal tubular interstitium and arteriosclerosis.Both SBP and losartan potassium had alleviated renal pathological change,and significantly reduced the infiltration of macrophage(P<0.05,P<0.01).SBP and losartan potassium decreased the expressions of TLR4,NF-κB,MCP-1,iNOS,and Arg-1.Conclusion:SBP significantly modified the early hypertensive renal injury by reducing inflammation,and the effect was similar to losartan potassium.