Tauroursodeoxycholic Acid Inhibited Apoptosis and Oxidative Stress in H2O2-Induced BMSC Death via Modulating the Nrf-2 Signaling Pathway: the Therapeutic Implications in a Rat Model of Spinal Cord Injury.

Spinal cord injury (SCI) is a prevalent and significant injury to the central nervous system, resulting in severe consequences. This injury is characterized by motor, sensory, and excretory dysfunctions below the affected spinal segment. Transplantation of bone marrow mesenchymal stem cells (BMSCs) has emerged as a potential treatment for SCI. However, the low survival as well as the differentiation rates of BMSCs within the spinal cord microenvironment significantly limit their therapeutic efficiency. Tauroursodeoxycholic acid (TUDCA), an active ingredient found in bear bile, has demonstrated its neuroprotective, antioxidant, and antiapoptotic effects on SCI. Thus, the present study was aimed to study the possible benefits of combining TUDCA with BMSC transplantation using an animal model of SCI. The results showed that TUDCA significantly enhanced BMSC viability and reduced apoptosis (assessed by Annexin V-FITC, TUNEL, Bax, Bcl-2, and Caspase-3) as well as oxidative stress (assessed by ROS, GSH, SOD, and MDA) both in vitro and in vivo. Additionally, TUDCA accelerated tissue regeneration (assessed by HE, Nissl, MAP2, MBP, TUJ1, and GFAP) and improved functional recovery (assessed by BBB score) following BMSC transplantation in SCI. These effects were mediated via the Nrf-2 signaling pathway, as evidenced by the upregulation of Nrf-2, NQO-1, and HO-1 expression levels. Overall, these results indicate that TUDCA could serve as a valuable adjunct to BMSC transplantation therapy for SCI, potentially enhancing its therapeutic efficacy.

Patchouli alcohol attenuates 5-fluorouracil-induced intestinal mucositis via TLR2/MyD88/NF-kB pathway and regulation of microbiota.

Intestinal mucositis causes great suffering to cancer patients who undergo chemotherapy and radiotherapy. Owing to the uncertain side effects of anticancer drugs to attenuate patients' intestinal mucositis, many studies focused on traditional Chinese medicine (TCM). Patchouli alcohol (PA) is an active compound extracted from Pogostemon cablin, and has potent gastrointestinal protective effect. However, whether PA has an effect on intestinal mucositis is still unknown. Therefore, we established a rat model of intestinal mucositis via intraperitoneal injection of 5-fluorouracil, and intragastrically administrated PA (10, 20, and 40 mg/kg) to evaluate the effect of PA on intestinal mucositis. The routine observation (body weight, food intake, and diarrhea) in rats was used to detect whether PA had an effect on intestinal mucositis. Levels of inflammatory cytokines (TNF-α, IL-1ß, IL-6, IL-10, and MPO), mucosal barrier proteins (zonula occludens -1 (ZO-1), claudin-1, occludin, myosin light chain (MLC), and mucin-2) and intestinal microbiota were determined to elucidate the underlying mechanism of PA action on intestinal mucositis in rats. The results showed that PA could effectively improve body weight, food intake, and diarrhea in intestinal mucositis rats, preliminary confirming PA efficacy. Further experiments revealed that PA not only decreased the levels of TNF-α, IL-1ß, IL-6, and MPO but also increased the level of IL-10 significantly. In addition, the expression of mucosal barrier proteins and microbiota community were also improved after PA treatment in diseased rats. Hence, PA may prevent the development and progression of intestinal mucositis by improving inflammation, protecting mucosal barrier, and regulating intestinal microbiota.

Protective role of ß-patchoulene from Pogostemon cablin against indomethacin-induced gastric ulcer in rats: Involvement of anti-inflammation and angiogenesis.

BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are most widely used as effective anti-inflammatory agents. However, their clinical application brings about inevasible gastrointestinal side effects. Pogostemon cablin is a traditional herbal medicine used for the treatment of gastrointestinal diseases in China. One of its representative components, the tricyclic triterpenoid ß-patchoulone (ß-PAE) has demonstrated great anti-inflammatory activity and gastroprotective effect against ethanol-induced gastric injury, but its protective effect against gastric ulcer induced by indomethacin is still unknown. PURPOSE: To assess the protective effect of ß-PAE against ulcer produced by indomethacin and reveal the underlying pharmacological mechanism. STUDY DESIGN: We used an indomethacin-induced gastric ulcer model of rats in vivo. METHODS: Gastroprotective activity of ß-PAE (10, 20, 40 mg/kg, i.g.) was estimated via indomethacin-induced gastric ulcer model in rats. Histopathological and histochemical assessment of ulcerated tissues were performed. Protein and mRNA expression were determined by Elisa, Western blotting and qRT-PCR. RESULTS: ß-PAE could inhibit ulcer formation. Histopathological and histochemical assessment macroscopically demonstrated that ß-PAE alleviates indomethacin-induced gastric ulceration in dose-dependent manner. After administration of ß-PAE, elevated tumor necrosis factor -α level was significantly decreased and the phosphorylation of JNK and IκB was markedly inhibited. ß-PAE suppressed the levels of E-selectin, P-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule and monocyte chemoattractant protein 1, as well as myeloperoxidase. Meanwhile, ß-PAE increased cyclooxygenase enzyme activities (COX-1 and COX-2) to enhance the production of prostaglandin E2. Proangiogenic protein, vascular endothelial growth factor and its receptor fms-like tyrosine kinase-1 mRNA expression were promoted while anti-angiogenic protein, endostatin-1 and its receptor ETAR mRNA expression were decreased. CONCLUSION: ß-PAE may provide gastroprotection in indomethacin-induced gastric ulcer in rats by reducing inflammatory response and improving angiogenesis.

Patchoulene Epoxide Isolated from Patchouli Oil Suppresses Acute Inflammation through Inhibition of NF-κB and Downregulation of COX-2/iNOS.

According to the GC-MS analysis, compositional variation was observed between samples of patchouli oil, of which an unknown compound identified as patchoulene epoxide (PAO) was found only in the long-stored oil, whose biological activity still remains unknown. Therefore, the present study aimed to evaluate the potential anti-inflammatory activity with three in vivo inflammatory models: xylene-induced ear edema, acetic acid-induced vascular permeability, and carrageenan-induced paw edema. Further investigation into its underlying mechanism on carrageenan-induced paw edema was conducted. Results demonstrated that PAO significantly inhibited the ear edema induced by xylene, lowered vascular permeability induced by acetic acid and decreased the paw edema induced by carrageenan. Moreover, PAO markedly decreased levels of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), prostaglandin E2 (PGE2), and nitric oxide (NO), but increased levels of interleukin-4 (IL-4) and interleukin-10 (IL-10). PAO was also shown to significantly downregulate the protein and mRNA expressions of cyclooxygenase-2 (COX-2) and inducible nitric-oxide synthase (iNOS). Western blot analysis revealed that PAO remarkably inhibited p50 and p65 translocation from the cytosol to the nucleus by suppressing IKKß and IκBα phosphorylation. In conclusion, PAO exhibited potent anti-inflammatory activity probably by suppressing the activation of iNOS, COX-2 and NF-κB signaling pathways.

[Comparative Study on Quality of Volatile Oil of Acori Tatarinowii Rhizoma from Two Habitats].

OBJECTIVE: To compare the quality of volatile oil of Acori Tatarinowii Rhizoma from Guangxi and Sichuan. METHODS: The volatile oil was extracted from Acori Tatarinowii Rhizoma by using steam distillation method, and analyzed by GC-MS. Peak area normalization method was used for calculating the relative percentage contents of chemical constituents, and hierarchical cluster analysis was used for classifying the 20 batches of samples by their relative contents of the main components that were methyleugenol, cis-methylisoeugenol, γ-asarone, ß-asarone and α-asarone. RESULTS: The average extraction rate of 10 batches of volatile oil in Acori Tatarinowii Rhizoma from Guangxi was 1. 61%, and 10 batches of samples from Sichuan was 1. 72%. The relative percentage contents of five main components totaled 78. 19% and 88. 84%, respectively. By t-test, there was no statistical difference between samples from Guangxi and Sichuan. In the hierarchical cluster analysis,10 batches of samples from Guangxi and 10 batches of samples from Sichuan could respectively be classified into four clusters subcategories and five clusters subcategories, while the mean of samples of Guangxi and the mean of samples of Sichuan respectively analyzed with 20 batches of the two habitats that all were classified in the same clusters subcategories. The results of similarity showed that the correlation coefficients of 8 batches in 10 batches of samples from Guangxi were over 0. 9, while 1 batch was only 0. 466. The correlation coefficients of 7 batches in 10 batches of samples from Sichuan had were over 0. 9, while 1 batch was only 0. 069. The correlation coefficients between the mean of samples of Guangxi and the mean of samples of Sichuan was 0. 996. CONCLUSION: Quality of the different batches of volatile oil from Acori Tatarinowii Rhizoma have significant differences, but it has no obvious correlation with the habitats.

[Analysis of the constituents of volatile oil from Fructus Auranti Immaturus by GC-MS].

OBJECTIVE: To analysis the constituents of volatile oil from Fructus Auranti Immaturus by GC-MS. METHODS: The volatile oil was extracted by steam distillation, then separated by capillary gas chromatography. The constituents of volatile oil were identified and their amount were determined by normalization method. RESULTS: Nineteen components were identified from Fructus Auranti Immaturus and their amount accounted 95.791% of total volatile oil. The main components were Limonene (68.25%), gamma-Terpinene (13.02%), alpha-Terpinol (3.28%), beta-Cymene (3.09%), beta-Myrcene (2.34%), alpha-Pinene (1.53%), beta-Pinene (1.05%). CONCLUSION: The main component of volatile oil from Fructus Auranti is Limonene.

[Effects of volatile oil of Rhizoma Acori Tatarinowii on morphology and cell viability in cultured cardiac myocytes].

OBJECTIVE: To observe the effects of volatile oil of Rhizoma Acori Tatarinowii (VOSCP) on morphology and cell viability in cultured neonate rat cardiac myocytes. METHODS: The cardiac myocytes were cultured by trypsin under cool treatment, and the cell purity was assayed with imminocytochemistry; Morphological changes were observed under phase contrast microscope after cardiac myocytes with VOSCP of different concentration for 24h in vitro, and the cell viability was examed by MTT assay. RESULTS: The purity of cultured neonate rat cardiac myocytes was higher than 95%; VOSCP of different concentration could depress pulse frequency and 100-160 mg/L VOSCP can obviously improve the viability of cardiac myocytes. The relation of dose-effect relationship was a parabola whose peak was at the value of 140 mg/L. CONCLUSION: VOSCP of proper concentration can depress pulse frequency and improve the viability of cardiac myocytes.

[Effects of beta-asaron on gene expression in mouse brain].

OBJECTIVE: To observe effects of beta-asaron on gene expression in BALB/c mouse brain by gene chip. METHODS: The mRNA of mouse brain samples of experiment group and control group was markered with cy5, cy3 fluorescence separately, and hybridized with 4000 genes on MGEC-40S gene chip after mix. Fluorescence intensity was scanned with Genepix 400 B scanner and analysed with Genepix Pro3.0 soft. RESULTS: Among 4000 genes, there are 15 genes with difference expression (the ratio > 1.9 or < 0.59) in two groups. In beta-asaron group, 4 genes expression raised relating to ion channels, substance transform through membrane, calcium-dependent protein kinase regulation, apoptosis, 11 genes expression declined relating to excitatory neurotransmitter metabolism in the CNS, T lymphocyte tend, genes expression regulation and drug metabolism. CONCLUSION: beta-asaron has effects on multi-target genes in mouse brain.