Modified Sijunzi Decoction Inhibits Epithelial-Mesenchymal Transition of Non-Small Cell Lung Cancer by Attenuating AKT/GSK3ß Pathway in vitro and in vivo.

Modified Sijunzi Decoction (MSJZD) is an empirical prescription of Traditional Chinese Medicine (TCM) and has been corroborated to be effective in multiple human diseases, but its role in non-small cell lung cancer (NSCLC) is enigmatic. Here we mainly analyze the function and mechanism of MSJZD in NSCLC. In this study, we used a method that coupled ultra-performance liquid chromatography to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to investigate the major constituents in MSJZD with positive and negative ion modes. Additionally, in in vitro experiments, the effects of serum-containing MSJZD on the biological behavior of NSCLC cells induced by TGF-ß1 were assessed by cell function experiments. Then, the influences of serum-containing MSJZD on epithelial-mesenchymal transition (EMT)-related markers were examined by immunofluorescence and western blot assays. Also, the AKT/GSK3ß pathway and apoptosis-related markers were estimated by western blotting. Tumor xenografts were generated by subcutaneously injecting A549 cells into BALB/c nude mice to determine the effects of MSJZD in vivo. We first analyzed the composition of MSJZD. In positive ion mode, 47 kinds of components were identified. In negative ion mode, 45 kinds of components were identified. We also found that TGF-ß1 contributed to inducing cell morphological changes and EMT progression. In vitro, surprisingly, cell proliferation, migration as well as invasion in NSCLC cells induced by TGF-ß1, could be weakened by serum-containing MSJZD, and apoptosis was intensified. Moreover, serum-containing MSJZD weakened EMT passage and AKT/GSK3ß pathway activation and induced apoptosis-related markers in NSCLC cells triggered by TGF-ß1. In vivo, we discovered that MSJZD attenuated the tumor growth, promoted histopathological damage, and induced apoptosis in A549 tumor-bearing mice. Importantly, MSJZD has also restrained the development of EMT, AKT/GSK3ß pathway, and TGF-ß1 expression levels in nude mice. These findings demonstrated that MSJZD significantly weakened NSCLC progression by modulating EMT and AKT/GSK3ß pathway.

Network pharmacology based investigation into the effect and mechanism of Modified Sijunzi Decoction against the subtypes of chronic atrophic gastritis.

Chronic atrophic gastritis (CAG) is a common digestive disease without specific treatment. According to syndrome differentiation, traditional Chinese medicine (TCM) classified it into different syndromes and has achieved significant therapeutic effects. In this study, immune repertoire sequencing techniques combined with symptom scores, electronic gastroscopy as well as pathologic changes were used to evaluate the effect and the underlying mechanism of Modified Sijunzi Decoction (MSD) in treating CAG. The results showed that MSD could relieve CAG symptoms, improve pathologic changes in CAG with fatigue and tiredness symptom, but with no help in CAG with reversal heat symptom. Moreover, MSD could regulate immune disorders in CAG with fatigue and tiredness symptom, and 7 TCR biomarkers were explored in CAG patients with immune disorders. All these results indicated that MSD is effective in treating CAG patients with fatigue and tiredness symptom by tonifying the spleen qi, suggesting that CAG treatment based on syndrome differentiation is reasonable.

Modified Sijunzi decoction can alleviate cisplatin-induced toxicity and prolong the survival time of cachectic mice by recovering muscle atrophy.

ETHNOPHARMACOLOGICAL RELEVANCE: Sijunzi decoction is a well-known traditional Chinese medicine (TCM) commonly used for invigorating vital energy and for the enhancement of immunity. Modified Sijunzi decoctions have been extensively used to treat cachexia and improve the quality of life of cancer patients undergoing chemotherapy. AIM OF THE STUDY: This study was aimed to provide comprehensive evidence for the anti-cachectic effect of a modified Sijunzi decoction (Zhen-Qi; ZQ-SJZ) and characterize its anti-cachectic mechanism, especially in cisplatin-induced muscle atrophy. MATERIALS AND METHODS: We employed a Lewis lung carcinoma (LLC)-induced cancer cachectic mouse model to demonstrate the anti-cachectic effect of ZQ-SJZ. Moreover, we provided an in vitro C2C12 myotube formation model to investigate the effect of ZQ-SJZ in hampering cisplatin-induced muscle atrophy. RESULTS: The administration of ZQ-SJZ can recover tumor- and/or cisplatin-induced body weight loss, intestinal mucosal damage, as well as forelimb grip strength and myofiber size. The administration of ZQ-SJZ also significantly prolonged the survival of LLC-induced cachectic mice under cisplatin treatment. Mechanistically, ZQ-SJZ increased the levels of myogenic proteins, such as myosin heavy chain (MyHC) and myogenin, and decreased the atrophy-related protein, atrogin-1, in cisplatin-treated C2C12 myotubes in vitro. In addition, cisplatin-induced mitochondria dysfunction could be hampered by the co-administration of ZQ-SJZ, by which it recovered the cisplatin-mediated decrease in PGC-1α and PKM1 levels. CONCLUSIONS: The administration of ZQ-SJZ can recover tumor- and/or cisplatin-induced cachectic conditions and significantly prolong the survival of LLC-induced cachectic mice under cisplatin treatment. The profound effect of ZQ-SJZ in hampering tumor- and/or cisplatin-induced cachexia may be due to its modulation of the mitochondrial function and subsequent myogenesis. Taken together, these results demonstrated the anti-cachectic mechanism of ZQ-SJZ and its potential use as a palliative strategy to improve the efficacy of chemotherapy.

Effects of modified Sijunzi decoction on the expression of p53 and Bcl-2 in burn-induced esophageal lesion / 中国生化药物杂志

Objective To establish burn-induced esophageal lesion model by adding NaOH with different concentrations in rabbits, and investigate the effect of modified sijunzi decoction on the p53 and Bcl-2.Methods After injection with different concentrations of NaOH, esophagus was dissected and observed anatomically.Rabbits were given a gavage of modified Sijunzi decoction daily for 20 days, and then were injected with NaOH.Esophageal epithelium isolated from each group was observed by hematoxylin-eosin staining.p53 and Bcl-2 protein and mRNA expression was measured with western blot and qRT-PCR, respectively.Results The degree of corrosion of esophageal epithelium was positively correlated with the concentration of NaOH.p53 protein and mRNA levels were increased after NaOH challenge; this increase was inhibited by treatment with modified Sijunzi decoction.Additionally, NaOH decreased Bcl-2 protein and mRNA, which was attenuated by modified Sijunzi decoction.Conclusion Modified sijunzi decoction can relieve the esophageal alkali burning in a dose-dependent manner, suggesting that modified Sijunzi decoction may be a useful strategy to treat chemical injuries in esophageal tissue.

Effect of Modified Sijunzi Decoction on Humoral Immunity of Rats with Exercise-induced Exhaustion / 广州中医药大学学报

Objective To observe the influence of modified Sijunzi Decoction on the decrease of immune function of exercise-induced exhaustion rats. Methods Thirty SD rats were given adaptive training, and then 6 dead or non-adaptive rats were dropped out. The remaining 24 SD rats were randomly divided into normal group ( N= 8) , model group ( N = 8) , and modified Sijunzi Decoction group ( MSD group, N = 8) . Normal group did not receive any medication and took diet freely. The rats of model group and MSD group were orally given distilled water or modified Sijunzi Decoction 8 g per day respectively after the modeling by exercise-induced exhaustion. After 61 days, the serum was taken from the three groups, and immunoglobulin lgA, lgG, lgM, and complement C3,C4 were detected. Results The contents of immunoglobulins IgA, IgG, IgM and complements C3, C4 in MSD group were higher than those in the model group ( P<0.01) . Conclusion The decrease of immunity occurs in the process of exercise stress, but can be delayed by modified Sijunzi Decoction..