Study on the Effective Material Basis and Mechanism of Traditional Chinese Medicine Prescription (QJC) Against Stress Diarrhea in Mice

Stress diarrhea is a major challenge for weaned piglets and restricts pig production efficiency and incurs massive economic losses. A traditional Chinese medicine prescription (QJC) composed of Astragalus propinquus Schischkin (HQ), Zingiber officinale Roscoe (SJ), and Plantago asiatica L. (CQC) has been developed by our laboratory and shows marked anti-stress diarrhea effect. However, the active compounds, potential targets, and mechanism of this effect remain unclear and warrant further investigation. In our study, we verified the bioactive compounds of QJC and relevant mechanisms underlying the anti-stress diarrhea effect through network pharmacology and in vivo experimental studies. After establishing a successful stress-induced diarrhea model, histomorphology of intestinal mucosa was studied, and Quantitative real-time PCR (RT-qPCR) probe was used for the phosphoinositide 3-kinase (PI3K)–Akt signaling pathway to verify the therapeutic effect of QJC on diarrhea. First, using the network pharmacology approach, we identified 35 active components and 130 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in QJC. From among these, we speculated that quercetin, luteolin, kaempferol, scutellarein, and stigmasterol were the main bioactive compounds and assumed that the anti-diarrhea effect of QJC was related to the PI3K–Akt signaling pathway. The RT-qPCR indicated that QJC and its bioactive components increased the expression levels of PI3K and Akt, inhibited the expression of phosphatase and tensin homolog (PTEN), and activated the PI3K–Akt signaling pathway to relieve stress-induced diarrhea. Furthermore, we found that QJC alleviated the pathological condition of small intestine tissue and improved the integrity of the intestinal barrier. Taken together, our study showed that the traditional Chinese medicine QJC, quercetin, luteolin, kaempferol, scutellarein, and stigmasterol alleviated the pathological condition of small intestine tissue and relieved stress-induced diarrhea by increasing the expression levels of PI3K and Akt and inhibiting the expression levels of PTEN.

Jianpi Qushi Heluo Formula alleviates renal damages in Passive Hemann nephritis in rats by upregulating Parkin-mediated mitochondrial autophagy

Jianpi Qushi Heluo Formula (JQHF) is an empirical traditional Chinese medicine prescription for treating Membranous Nephropathy (MN) clinically in China. The therapeutic effect of JQHF has been reported in our previous studies. However, the exact mechanism is still unknown. In this study, by establishing an experimental rat model of MN induced by Sheep anti-rat Fx1A serum, we evaluated the effects of JQHF and Tetrandrine (TET), and Benazepril was used as a positive control. As an autophagy agonist, TET is one of the most active components in JQHF. After 4 weeks, significant kidney damage was observed in the rats in the Model group; comparatively, JQHF markedly decreased 24 h urinary protein, Total Cholesterol (TC), and increased serum total Albumin (ALB). Histology showed that JQHF caused significant improvements in glomerular hyperplasia, renal tubular damage, IgG immune complex deposition, and the ultrastructure of mitochondria in MN rats. Flow cytometry analysis showed that treatment with JQHF reduced the level of reactive oxygen species and apoptosis rate, and upregulated mitochondrial membrane potential. Western blot analysis demonstrated that JQHF could protect against mitochondrial dysfunction and apoptosis by upregulating the expression of PINK1, Mitochondrial Parkin, and LC3-II/I, downregulating the expression of Cytoplasmic Parkin, P62, Cytochrome c, and Caspase-3 in the kidneys of MN rats. From images of co-immunofluorescence, it is observed significantly increase in the co-localization of PINK1 and Parkin, as well as LC3 and mitochondria. Similarly, TET treatment significantly upregulated the mitochondrial autophagy and reduced apoptosis in rats after 4 weeks compared with the model group. Comparatively, the ability of JQHF to alleviate renal damage was significantly higher than those of Benazepril and TET. It was demonstrated that JQHF could delay pathology damage to the kidney and hold back from the progression of MN by inhibiting apoptosis and upregulating the mitochondrial autophagy by PINK1/Parkin pathways.

A Network Pharmacology-Based Study of the Mechanism of Renshen Baidu Powder in Treating Ulcerative Colitis

Ulcerative colitis (UC), one forms of chronic inflammatory bowel disease (IBD), is characterized by recurrent inflammation, and Renshen Baidu Powder (RSBDP) is often used as a traditional Chinese medicine prescription for treating of UC, but the pharmacological mechanisms remains unclear. This research aims to explore the mechanisms of RSBDP in the treatment of UC based on network pharmacology. Through multiple databases collected the compounds and targets, and constructed the network. The crossover genes enrichment analysis were performed by protein-protein interaction (PPI) construction and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, 181 compounds and 266 targets were obtained. Potential candidate ingredients mainly include phytosterols, flavonoids, and coumarins, all of which have anti-inflammatory activities. GO and KEGG enrichment analysis suggested that RSBDP have anti-inflammatory and immunomodulatory effects. The effect of RSBDP on UC might be achieved by regulating the balance of cytokines (i.e., IL-6, TNF, IL-1) and the downstream mediators (i.e., STAT1, STAT3) in the immune system. And inflammation-, immune- and hypoxic-related pathways, like TNF, Toll-like receptors and HIF-1 signaling pathway. These results provide a theoretical basis for studying the effective substances of RSBDP in the treatment of UC and their mechanism of action for further research.

Sheng Jing Decoction, as a Traditional Chinese Medicine Prescription, Can Promote Spermatogenesis and Increase Sperm Motility

BackgroundSheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. It has been applied in many hospitals in China, and the clinical effect of patients' reaction is satisfactory. However, pharmacological function and molecular mechanism of SJD are poorly understood. In this study, mainly investigated the function of SJD on spermatogenesis and sperm motility, and explored the potential mechanisms.MethodsThe oligozoospermia model of ICR mice was induced by injecting intraperitoneally (ip) with 60 mg/kg dose of cyclophosphamide. At the same time of modeling, high, medium and low doses of SJD were given orally for treatment respectively. Sperm vitality and motility were detected and analyzed by CASA, and sperm morphology was tested by Papanicolaou staining. Sperm mitochondrial membrane potential (MMP) was tested by JC-1 staining. Sperm plasma membrane integrity was determined by SYBR-14/PI double staining. Histopathological changes of testis were analyzed by HE staining and Immunohistochemistry. Genes expression related to spermatogenesis and sperm development were detected by real-time RT-PCR.Results High, medium and low doses of SJD are effective to recover from the impairment of the whole body and testicular tissue by cyclophosphamide inducing, and to rescue the damage of testicular tissue cells including sertoli cells and germ cells by cyclophosphamide inducing. SJD can all partly restore the decrease in sperm concentration, sperm vitality, sperm motility and normal sperm morphology rate by cyclophosphamide inducing. Ki67 staining analyses confirm SJD can promoted testicular tissue cells proliferation. Real-time RT-PCR analyses reveal SJD can up-regulates the expression of proliferation-associated gene Lin28a, and differentiation-associated genes Kit, Sohlh2 and Stra8. SJD can also reduce the impairment of MMP and sperm plasma membrane integrity by cyclophosphamide inducing.ConclusionSJD is effective to support sperm quantity and quality by increasing sperm concentration, sperm vitality, sperm motility and normal sperm morphology rate. SJD can promote spermatogenesis by up-regulating the expression of the proliferation-associated gene Lin28a, and the differentiation-associated genes (Kit, Sohlh2 and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.