A bioinformatics and network pharmacology approach to the mechanisms of action of Shenxiao decoction for the treatment of diabetic nephropathy.

BACKGROUND: The epithelial-to-mesenchymal transition (EMT) of renal tubular epithelial cells is the main pathological alteration in diabetic nephropathy (DN). Traditional Chinese medicine (TCM) has been used for the treatment of DN in clinical practice and has been proven to be effective. PURPOSE: This aim of this study was to shed light on the efficacy of Shenxiao decoction (SXD) on the EMT of renal tubular epithelial cells and the molecular mechanisms of SXD in mice with DN, as well as on the high glucose (HG)- and TGF-ß1-induced EMT of NRK-52E and HK-2 cells. STUDY DESIGN AND METHODS: A bioinformatics and network pharmacology method were utilized to construct the active ingredient-target networks of SXD that were responsible for the beneficial effects against DN. The effects of RUNX3 were validated in HG- and TGF-ß1-induced EMT processes in NRK-52E and HK-2 cells. RESULTS: Bioinformatics analysis revealed that 122 matching targets were closely associated with the regulation of cell migration and the AGE-RAGE signaling pathway in diabetic complications. The results also revealed that, relative to the mice with DN, the mice in the treatment group had an improved general state and reduced blood glucose levels. The degradation of renal function was ameliorated by SXD. Moreover, the protective effects of SXD were also observed on renal structural changes. Furthermore, SXD suppressed the activation of the transforming growth factor (TGF)-ß1/Smad pathway and upregulated the RUNX3 and E-cadherin levels and downregulated the extracellular matrix (ECM) protein levels in mice with DN. SXD was further found to prevent the HG- and TGF-ß1-induced EMT processes in NRK-52E and HK-2 cells. Additionally, the overexpression of RUNX3 markedly inhibited the EMT and TGF-ß1/Smad pathway induced by HG and TGF-ß1 in NRK-52E and HK-2 cells. CONCLUSION: Taken together, these results suggest that SXD maybe alleviate EMT in DN via the inhibition of the TGF-ß1/Smad/RUNX3 signaling pathway under hyperglycemic conditions.

[Mechanism of effective components of Mori Folium in alleviating insulin resistance based on JNK signaling pathway].

A stable hepatoma cell line(Hep G2 cell) insulin resistance model was established and used to analyze the effect of effective components of Mori Folium in alleviating insulin resistance,and preliminary explore the mechanism for alleviating insulin resistance. The Hep G2 insulin action concentration and the duration of action were investigated using the glucose oxidase method(GOD-POD method) to establish a stable Hep G2 insulin resistance model. Normal control group,model group,Mori Folium polysaccharide group,Mori Folium flavonoid group and rosiglitazone group were divided to determine the glucose consumption. The effect of Mori Folium effective components on Hep G2 insulin resistance was analyzed. The mRNA expressions of JNK,IRS-1 and PDX-1 in each group were detected by Real-time quantitative PCR(qRT-PCR). The protein expressions of p-JNK,IRS-1 and PDX-1 were detected by Western blot. And the mechanism of effective components of Mori Folium in alleviating insulin resistance was investigated. The results showed that the glucose consumption was significantly decreased in the insulin resistance cells after incubation with 25. 0 mg·L-1 insulin for 36 h(P<0. 01),and the model was relatively stable within 36 h. Mori Folium polysaccharides and flavonoids all alleviated insulin resistance,among which Mori Folium flavonoids had better effect in alleviating Hep G2 insulin resistance(P<0. 05). The qRT-PCR analysis showed that Mori Folium polysaccharides and flavonoids could inhibit JNK and IRS-1 mRNA expressions,while enhancing PDX-1 mRNA expression. Western blot analysis displayed that Mori Folium polysaccharides and flavonoids could inhibit p-JNK and IRS-1 protein expressions,while enhancing PDX-1 protein expression. Mori Folium polysaccharides and flavonoids can alleviate insulin resistance in Hep G2 cells,and its mechanism may be the alleviation of insulin resistance by inhibiting JNK signaling pathway.

[Clinical observation on delayed encephalopathy after carbon monoxide poisoning treated with acupuncture to restore consciousness combined with hyperbaric oxygen treatment].

OBJECTIVE: To compare the efficacy differences on delayed encephalopathy after carbon monoxide poisoning (DEACMP) between acupuncture to restore consciousness combined with hyperbaric oxygen treatment and simple hyperbaric oxygen treatment. METHODS: Forty-one patients with DEACMP were randomly divided into an observation group (21 cases) and a control group (20 cases). In the observation group, acupuncture was applied at Neiguan (PC 6), Shuigou (GV 26), Baihui (GV 20), Sishencong (EX-HN 1), Fengchi (GB 20), Hegu (LI 4), Sanyinjiao (SP 6) and Taichong (LR 3), and hyperbaric oxygen treatment was given as well. In the control group, simple hyperbaric oxygen treatment was used. The treatment was adopted once every day, and continuous 5 days' treatment made one session in the two groups. There were two days at the interval between two sessions and 6 sessions in the two groups. The changes of scores of mini mental state examination (MMSE) and Barthel index (BI) for activity of daily life and routine electroencephalogram (EEG) before and after treatment were compared. RESULTS: After treatment, the scores of MMSE and BI and EEG were all improved compared with those before treatment (all P<0.01). The raise of the scores of MMSE and BI in the observation group was more obvious than that in the control group (both P<0.05) and the improvement of EEG abnormal condition in the observation group was also superior to that in the control group after treatment (P<0.05). CONCLUSION: Acupuncture to restore consciousness combined with hyperbaric oxygen could obviously improve the cognitive function, activity of daily life and changes of EEG, and it is better than simple hyperbaric oxygen treatment.

Latent variable analysis on Chinese medicine syndrome in patients with antituberculosis drug-induced liver injury.

OBJECTIVE: To explore the Chinese medicine (CM) syndrome discipline in patients with antituberculosis drug (ATBD)-induced liver injury to provide the basis of the standard Chinese medicine treatment for the disease with latent variable analysis. METHODS: Epidemiological investigation method was adopted. Two hundred and sixty-one patients with ATBD-induced liver injury were investigated using CM syndrome questionnaire. The syndrome types were determined with exploratory factor analysis (EFA), and the latent variables were analyzed by confirmatory factor analysis (CFA). RESULTS: Totally 26 indexes related to CM syndrome differentiation were obtained from the 261 eligible cases, among them, 5 were as the latent dependent variables, which corresponded to 5 common syndrome types, including dampness encumbering the Spleen (Pi), Liver (Gan)-qi stagnation, Spleen and Stomach (Wei) deficiency, stasis-toxin accumulation, and qi-yin deficiency. CFA indicated that the indexes with loading coefficient [Symbol: see text]0.6 exactly reflected the connotation of its corresponding syndrome type. CONCLUSIONS: Five CM syndrome types are the most common in patients with ATBD-induced liver injury, which relate to their corresponding indexes for differentiation. It is feasible to apply combined EFA and CFA for explaination and measurement of the existence of CM syndrome under specific diseases.

Study on the relationship between Chinese medicine constitutive susceptibility and diversity of syndrome in diabetic nephropathy.

OBJECTIVE: To explore the relationship between Chinese medicine (CM) constitutive susceptibility and syndrome diversity in diabetic nephropathy (DN). METHODS: Epidemiologic investigation on constitution adopting the "Constitution in Chinese Medicine Questionnaire" (CCMQ), and survey on syndrome type by CM syndrome scale (preliminary) were carried out in 180 DN patients. Cluster analysis on symptom items was used to determine the syndrome type, and canonical correlation analysis was used to analyze the relationship between patients' constitution and syndrome. RESULTS: Baseline levels in all enrolled patients were not different statistically. Cluster analysis showed 8 syndromes existed in DN patients, namely: I, qi-yin deficiency with qi-stagnancy type; II, yin-yang deficiency with heat-water-blood stasis type; III, qi-yin deficiency with dampness-heat type; IV, yin-yang deficiency with blood-stasis and heat type; V, qi-yin deficiency with stagnant heat type; VI, yin-yang deficiency with inner dampness-heat stagnancy type; VII, yin deficiency with heat stagnancy type; and VIII, Kidney (Shen)-Spleen (Pi) deficiency with stagnant heat type. Correlation analysis on the 8 syndromes and the 9 constitutions showed statistical significant correlations between syndrome III and dampness-heat constitution (P=0.0001); syndrome IV and blood-stasis constitution (P=0.0001); and syndrome VII and yin-deficiency constitution (P=0.0180). CONCLUSION: Certain relationship revealed between CM constitutions and syndrome types; constitution decides the disease genesis, its syndrome type and prognosis, as well as the change of syndromes.

Association of Chinese medicine constitution susceptibility to diabetic nephropathy and transforming growth factor-ß1 (T869C) gene polymorphism.

OBJECTIVE: To explore the association of Chinese medicine constitution susceptibility to diabetic nephropathy (DN) and transforming growth factor (TGF)-ß1 (T869C) gene polymorphism. METHODS: TGF-ß1 gene polymorphism detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was screened for 180 DN cases and 180 type 2 diabetic mellitus (T2DM) cases without combined DN. Patients with DN were surveyed epidemiologically with constitution in the Chinese medicine questionnaire (CCMQ). Binary logistic regression analysis was utilized to study the correlation between nine types of Chinese medicine constitution and TGF-ß1 (T869C) gene polymorphisms. RESULTS: The DN group has a higher frequency of TGF-ß1 (T869C) gene polymorphism than the T2DM group, and CC/CT genotypes than the T2DM group [CC, CT, TT (DN group): 88, 87, 5 (cases) versus (T2DM group) 71, 73, 36 (cases), P<0.05]. The phlegm-dampness constitution, damp-heat constitution, and blood stasis constitution have correlations with TGF-ß1 (T869C) gene polymorphism. CONCLUSION: Chinese medicine constitutions were associated with TGF-ß1 (T869C) gene polymorphism, a potential predictor of susceptibility to DN in T2DM patients.