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Molecular signatures in the prevention of radiation damage by the synergistic effect of N-acetyl cysteine and qingre liyan decoction, a traditional chinese medicine, using a 3-dimensional cell culture model of oral mucositis.

Lambros MP, Kondapalli L, Parsa C, Mulamalla HC, Orlando R, Pon D, Huang Y, Chow MS - Evid Based Complement Alternat Med (2015)

Bottom Line: Microarray results revealed that the NAC-QYD caused the upregulation of genes encoding metallothioneins, HMOX1, and other components of the Nrf2 pathway, which protects against oxidative stress.Finally, tissue-cultures treated prophylactically with NAC-QYD showed significant downregulation of apoptosis, cytokines and chemokines genes, and constrained damage-associated molecular patterns (DAMPs).NAC-QYD treatment involves the protective effect of Nrf2, NFκB, and DNA repair factors.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Western University of Health Sciences, Pomona, CA 91766, USA.

ABSTRACT
Qingre Liyan decoction (QYD), a Traditional Chinese medicine, and N-acetyl cysteine (NAC) have been used to prevent radiation induced mucositis. This work evaluates the protective mechanisms of QYD, NAC, and their combination (NAC-QYD) at the cellular and transcriptional level. A validated organotypic model of oral mucosal consisting of a three-dimensional (3D) cell tissue-culture of primary human keratinocytes exposed to X-ray irradiation was used. Six hours after the irradiation, the tissues were evaluated by hematoxylin and eosin (H and E) and a TUNEL assay to assess histopathology and apoptosis, respectively. Total RNA was extracted and used for microarray gene expression profiling. The tissue-cultures treated with NAC-QYD preserved their integrity and showed no apoptosis. Microarray results revealed that the NAC-QYD caused the upregulation of genes encoding metallothioneins, HMOX1, and other components of the Nrf2 pathway, which protects against oxidative stress. DNA repair genes (XCP, GADD45G, RAD9, and XRCC1), protective genes (EGFR and PPARD), and genes of the NFκB pathway were upregulated. Finally, tissue-cultures treated prophylactically with NAC-QYD showed significant downregulation of apoptosis, cytokines and chemokines genes, and constrained damage-associated molecular patterns (DAMPs). NAC-QYD treatment involves the protective effect of Nrf2, NFκB, and DNA repair factors.

No MeSH data available.


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H and E staining of 3D-cultured human oral tissue. Nonirradiated, nontreated oral epithelium (negative control) consists of human oral keratinocytes and is divided into the stratum basalis (basal layer), stratum spinosum (the spindle shaped cell layer), and the stratum corneum (a); irradiated, nontreated tissue (positive control). Keratinocyte injury is noticeable at the top of the tissues, with an increased number of pyknotic cells at the bottom of the tissue (b); irradiated tissue prophylactically treated with NAC shows keratinocyte injury at the top of the tissue (c); irradiated tissue prophylactically treated with QYD shows keratinocyte injury (d); irradiated tissue prophylactically treated with NACQYD shows no injury and resembles the nonirradiated, nontreated negative control tissues (e).
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fig1: H and E staining of 3D-cultured human oral tissue. Nonirradiated, nontreated oral epithelium (negative control) consists of human oral keratinocytes and is divided into the stratum basalis (basal layer), stratum spinosum (the spindle shaped cell layer), and the stratum corneum (a); irradiated, nontreated tissue (positive control). Keratinocyte injury is noticeable at the top of the tissues, with an increased number of pyknotic cells at the bottom of the tissue (b); irradiated tissue prophylactically treated with NAC shows keratinocyte injury at the top of the tissue (c); irradiated tissue prophylactically treated with QYD shows keratinocyte injury (d); irradiated tissue prophylactically treated with NACQYD shows no injury and resembles the nonirradiated, nontreated negative control tissues (e).

Mentions: Using a 3D human cell culture model of oral keratinocytes, we studied the protective effect of three treatments on radiation damage. H and E staining of the nonirradiated tissue revealed a healthy, well-differentiated multilayer epithelium consisting of keratinocytes. In the lower part of the epithelium, the stratum basalis and stratum spinosum layers, consisting of cylindrical cells and elongated spindle cells, respectively, could be distinguished (Figure 1(a)). Untreated tissue samples irradiated with 12 Gy showed laceration and damage to the top part of the tissues (Figure 1(b)) and the presence of apoptotic cells (Figure 2(b)). Tissues pretreated with NAC before irradiation also revealed the formation of edematous and apoptotic cells (Figures 1(c) and 2(c)). The pretreatment of tissues with QYD showed fewer edematous cells (Figure 1(d)) compared with those pretreated with NAC (Figure 1(c)); however, the pretreatment with NAC-QYD resulted in healthy 3D tissue (Figure 1(e)) similar to the findings in the nonirradiated tissue.


Molecular signatures in the prevention of radiation damage by the synergistic effect of N-acetyl cysteine and qingre liyan decoction, a traditional chinese medicine, using a 3-dimensional cell culture model of oral mucositis.

Lambros MP, Kondapalli L, Parsa C, Mulamalla HC, Orlando R, Pon D, Huang Y, Chow MS - Evid Based Complement Alternat Med (2015)

H and E staining of 3D-cultured human oral tissue. Nonirradiated, nontreated oral epithelium (negative control) consists of human oral keratinocytes and is divided into the stratum basalis (basal layer), stratum spinosum (the spindle shaped cell layer), and the stratum corneum (a); irradiated, nontreated tissue (positive control). Keratinocyte injury is noticeable at the top of the tissues, with an increased number of pyknotic cells at the bottom of the tissue (b); irradiated tissue prophylactically treated with NAC shows keratinocyte injury at the top of the tissue (c); irradiated tissue prophylactically treated with QYD shows keratinocyte injury (d); irradiated tissue prophylactically treated with NACQYD shows no injury and resembles the nonirradiated, nontreated negative control tissues (e).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4326035&req=5

fig1: H and E staining of 3D-cultured human oral tissue. Nonirradiated, nontreated oral epithelium (negative control) consists of human oral keratinocytes and is divided into the stratum basalis (basal layer), stratum spinosum (the spindle shaped cell layer), and the stratum corneum (a); irradiated, nontreated tissue (positive control). Keratinocyte injury is noticeable at the top of the tissues, with an increased number of pyknotic cells at the bottom of the tissue (b); irradiated tissue prophylactically treated with NAC shows keratinocyte injury at the top of the tissue (c); irradiated tissue prophylactically treated with QYD shows keratinocyte injury (d); irradiated tissue prophylactically treated with NACQYD shows no injury and resembles the nonirradiated, nontreated negative control tissues (e).
Mentions: Using a 3D human cell culture model of oral keratinocytes, we studied the protective effect of three treatments on radiation damage. H and E staining of the nonirradiated tissue revealed a healthy, well-differentiated multilayer epithelium consisting of keratinocytes. In the lower part of the epithelium, the stratum basalis and stratum spinosum layers, consisting of cylindrical cells and elongated spindle cells, respectively, could be distinguished (Figure 1(a)). Untreated tissue samples irradiated with 12 Gy showed laceration and damage to the top part of the tissues (Figure 1(b)) and the presence of apoptotic cells (Figure 2(b)). Tissues pretreated with NAC before irradiation also revealed the formation of edematous and apoptotic cells (Figures 1(c) and 2(c)). The pretreatment of tissues with QYD showed fewer edematous cells (Figure 1(d)) compared with those pretreated with NAC (Figure 1(c)); however, the pretreatment with NAC-QYD resulted in healthy 3D tissue (Figure 1(e)) similar to the findings in the nonirradiated tissue.

Bottom Line: Microarray results revealed that the NAC-QYD caused the upregulation of genes encoding metallothioneins, HMOX1, and other components of the Nrf2 pathway, which protects against oxidative stress.Finally, tissue-cultures treated prophylactically with NAC-QYD showed significant downregulation of apoptosis, cytokines and chemokines genes, and constrained damage-associated molecular patterns (DAMPs).NAC-QYD treatment involves the protective effect of Nrf2, NFκB, and DNA repair factors.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Western University of Health Sciences, Pomona, CA 91766, USA.

ABSTRACT
Qingre Liyan decoction (QYD), a Traditional Chinese medicine, and N-acetyl cysteine (NAC) have been used to prevent radiation induced mucositis. This work evaluates the protective mechanisms of QYD, NAC, and their combination (NAC-QYD) at the cellular and transcriptional level. A validated organotypic model of oral mucosal consisting of a three-dimensional (3D) cell tissue-culture of primary human keratinocytes exposed to X-ray irradiation was used. Six hours after the irradiation, the tissues were evaluated by hematoxylin and eosin (H and E) and a TUNEL assay to assess histopathology and apoptosis, respectively. Total RNA was extracted and used for microarray gene expression profiling. The tissue-cultures treated with NAC-QYD preserved their integrity and showed no apoptosis. Microarray results revealed that the NAC-QYD caused the upregulation of genes encoding metallothioneins, HMOX1, and other components of the Nrf2 pathway, which protects against oxidative stress. DNA repair genes (XCP, GADD45G, RAD9, and XRCC1), protective genes (EGFR and PPARD), and genes of the NFκB pathway were upregulated. Finally, tissue-cultures treated prophylactically with NAC-QYD showed significant downregulation of apoptosis, cytokines and chemokines genes, and constrained damage-associated molecular patterns (DAMPs). NAC-QYD treatment involves the protective effect of Nrf2, NFκB, and DNA repair factors.

No MeSH data available.


Related in: MedlinePlus