Periareolar approach in video-assisted thoracoscopic surgery for right middle lobectomy: a novel technique.

BACKGROUND: Uniportal thoracoscopic right middle lobectomy (RML) poses greater technical challenges than other lobectomies. Although two-port thoracoscopy offers convenience, it results in heightened surgical trauma and scarring. The periareolar incision is rarely used in lobectomy while known for its cosmetic advantages. This study presents the periareolar access (combining a periareolar port and a 1-cm port) for video-assisted thoracoscopic surgery (VATS) in RML, comparing it with the traditional uniportal technique in both male and female patients. METHODS: Eighty patients who underwent RML were randomly divided into two groups: the periareolar VATS (PV) approach (n = 40) and the uniportal VATS (UV) approach (n = 40) from August 2020 to February 2023. All patients were followed up for 1 year and clinical data were collected and analyzed. RESULTS: No significant differences in complications, blood loss, duration of chest tube placement, and length of postoperative hospital stay were observed between two methods. However, the PV group exhibited significantly shorter operative time, reduced postoperative visible scarring and lower visual analogue scores (VAS) for postoperative pain (P < 0.05). Additionally, the PV group demonstrated significantly higher cosmetic and satisfaction scores at the 6-month postoperative assessment (P < 0.05). Notably, breast ultrasound follow-up revealed two cases injuries of the mammary glands in female patients, and sensory function of most nipple and areola remained intact except two cases in all PV group patients. CONCLUSIONS: Periareolar VATS emerges as a promising alternative approach for RML, providing clear benefits in pain management and cosmetic outcomes, while maintaining safety and convenience.

Reconstruction of the esophagus of patients with middle thoracic esophageal carcinoma using the remnant stomach following Billroth II gastrectomy.

It seems impossible to reconstruct the esophagus of patients with middle thoracic esophageal carcinoma with a history of distal gastrectomy using the remnant stomach. Although surgeons have made multiple efforts to reconstruct the esophagus using the remnant stomach, it can only be successfully used in cases of lower thoracic esophageal cancer. Additionally, the surgery is more complex than traditional esophagogastrostomy due to challenges including mobilization of the remnant stomach with the spleen and transposition of the pancreatic tail into the left hemithorax. Our operation proved that the remnant stomach, which we named as the completely mobilized remnant stomach after dissection of the feeding vessels, remained viable. We successfully integrated the completely mobilized remnant stomach in the reconstruction of the lower thoracic esophageal tract and then integrated it in Ivor Lewis esophagogastrostomy. We describe this new alternative surgical technique for the treatment of middle thoracic esophageal carcinoma in patients with a history of distal gastrectomy in this study. Clinical data of 23 patients from 2008 to 2019 were retrospectively analyzed. All patients underwent the Ivor Lewis procedure. All remaining vessels of the remnant stomach were dissected at their origins, and Roux-en-Y reconstruction or Braun anastomosis was performed. After esophagectomy during right thoracotomy, anastomosis of the remnant stomach and esophagus was performed. Two-field lymph node dissections were performed. There was no case of necrosis of the remnant stomach or of perioperative death. Serious complications included anastomotic leak in three cases, afferent-efferent loop syndrome in one, and anastomotic stricture in two. Application of the completely mobilized remnant stomach in Ivor Lewis esophagogastrostomy is feasible, and the surgical procedure is similar to that of normal esophagogastrostomy.

Modified left subsuperior segmentectomy via 2-cm uniportal video-assisted thoracoscopic surgery.

Independent subsuperior segmentectomy (S*) via uniportal video-assisted thoracoscopic surgery (VATS) has rarely been reported. We describe our modified technique of performing simplified left subsuperior segmentectomy for a lung nodule, via 2-cm uniportal VATS. The uniportal approach was different from the traditional approach made by blunt separation into the thorax without electrocautery. Our modified technique minimizes damage to the intercostal nerves and muscles. We also simplified the subsuperior segmentectomy procedure according to the findings of three-dimensional (3D) computed tomography angiography and bronchography. Combining these two techniques achieves a new more minimally invasive method for subsuperior segmentectomy.

Modified 2-cm super single port vs. the traditional 3-cm single port for video-assisted thoracoscopic surgery lobectomy.

PURPOSES: We introduce a novel 2-cm single port designed to minimize intercostal muscle and nerve damage in video-assisted thoracoscopic surgery (VATS) lobectomy, and compared it with the 3-cm traditional single port. METHODS: We analyzed, retrospectively, the clinical data, safety, convenience, incision complications, and postoperative pain and numbness in 81 patients who underwent either modified (n = 42) or traditional (n = 39) single-port VATS lobectomy. RESULTS: The preoperative variables were comparable between both single-port VATS lobectomy groups after matching. There were no serious complications and there was no mortality in either group. There were no remarkable differences between the groups in intraoperative blood loss, chest tube duration, lymph node dissection, or postoperative complications. The modified single-port group had a longer operation time (p < 0.05), but the static and dynamic postoperative VAS scores and incisional numbness were better in the modified single-port group (p < 0.05). The modified single-port group also had an obvious advantage in incision seepage, healing, and appearance. CONCLUSIONS: Our 2-cm modified single port for lobectomy is safe and effective, and results in less postoperative pain and incisional numbness than the 3-cm traditional single port.

Autophagy decreases alveolar epithelial cell injury by regulating the release of inflammatory mediators.

To research the impact of autophagy on alveolar epithelial cell inflammation and its possible mechanism in the early stages of hypoxia, we established a cell hypoxia-reoxygenation model and orthotopic left lung ischemia-reperfusion model. Rat alveolar epithelial cells stably expressing GFP-LC3 were treated with an autophagy inhibitor (3-MA) or an autophagy promoter (rapamycin), followed by hypoxia-reoxygenation treatment for 2, 4, and 6 hr in vitro. In vivo, 20 male Sprague Dawley rats were randomly divided into four groups (model group: No blocking of the hilum in the left lung; control group: Blocking of the hilum in the left lung for 1 hr with dimethyl sulfoxide lavage; 3-MA group: Blocking of the hilum in the left lung for 1 hr with 100 ml/kg of 3-MA (5 µmol/L) solution lavage; and rapamycin group: Blocking of the hilum in the left lung for 1 hr with 100 ml/kg of rapamycin (250 nmol/L) solution lavage) to establish an orthotopic left lung ischemia model. This study demonstrated that rapamycin significantly suppressed the nuclear factor kappa B signaling pathway and limited the expression of proinflammatory factors. A contrary result was found after the 3-MA pretreatment. These findings indicate that autophagy reduces ischemia-reperfusion injury by repressing inflammatory signaling pathways in the early stages of hypoxia in vitro and in vivo. Autophagy could be a new protective method for application in lung ischemia-reperfusion injury.

Proteasome inhibition promotes autophagy and protects from endoplasmic reticulum stress in rat alveolar macrophages exposed to hypoxia-reoxygenation injury.

Alveolar macrophages play vital roles in acute lung injury, and macrophage response to hypoxia play relevant roles to disease mechanisms. There is growing evidence that cell death pathways play crucial roles in physiological and pathological settings and that the ubiquitin-proteasome system is involved in the regulation of these processes. However, the functional role of proteasome in alveolar macrophages exposed to hypoxia-reoxygenation (H/R) injury is unknown. We aimed to investigate the function of proteasome on alveolar macrophages exposed to H/R and the underlying mechanisms. NR8383 cells were pretreated with proteasome activator sulforaphane (SFN) or inhibitor MG-132 for 1 hr, and then submitted to 2/6 hr, 4/6 hr, and 6/6 hr H/R treatment. Cell viability was assessed with MTT assay. Autophagy was monitored using electron transmission microscope and flow cytometry and western blotting. The endoplasmic reticulum (ER) stress and unfolded protein response (UPR) pathways were equally analyzed by western blotting. Cell apoptosis was detected by immunohistochemistry, caspase3/7 activity, and western blotting. The viability of NR8383 cells exposed to H/R was affected by proteasome activity and proteasome inhibition significantly inhibited cell death. Treatment with MG-132 led to autophagy activation and induced the survival of NR8383 cells exposed to H/R. Pretreatment with SFN significantly decreased cell autophagy and induced cell death. ER stress was activated in H/R-treated NR8383 cells, and SFN further promoted ER stress whereas proteasome inhibition led to contrary results. Proteasome inhibtion hindered cell apoptosis as demonstrated by decreased caspase-3/7 activity, immunolabelling, and western blot results. Proteasome inhibition might be a promising approach for treating H/R injury-related lung diseases.

Autophagy Activation by Rapamycin Before Hypoxia-Reoxygenation Reduces Endoplasmic Reticulum Stress in Alveolar Epithelial Cells.

BACKGROUND: To determine potential effects of autophagy activation on hypoxia-reoxygenation (H/R) induced damage of a rat alveolar epithelial cell line. METHODS: CCL149 cells were subjected to autophagy agonist (rapamycin, Rap), autophagy inhibitor (3-methyladenine, 3-MA) or PBS for 1 h before H/R treatment for 2 h, 4 h and 6 h. The optimal concentration of Rap (150 nM, 200 nM and 250 nM) or 3-MA (5 mM, 10 mM and 15 mM) was obtained from MTT assay. Autophagy was determined by fluorescence microscopy of eRFP-LC3 positive cells, transmission electron microscopy of autophagosome, western blot of LC3, AMPK, Beclin-1, HDAC6 and p62 proteins. Endoplasmatic reticulum stress was indicated by detecting expressions of BIP, XBP-1 and CHOP via western blot. RESULTS: Rap at concentration of 250 nM before H/R increased the autophagy formation with more eRFP-LC3 positive cells and higher expressions of LC3-II, Beclin-1, HDAC6 and p62, but lower expressions of BIP, XBP-1 and CHOP in H/R treated CCL149. This effect seemed to be still obvious after H/R exposure for 6 h. The contrary results were obtained by treatment with 5 mM 3-MA. CONCLUSION: Rap might be a promising agent before mechanical ventilation or reperfusion to prevent re-damage in hypoxia related lung diseases.

Blocking the NOTCH pathway can inhibit the growth of CD133-positive A549 cells and sensitize to chemotherapy.

Cancer stem cells (CSCs) are believed to play an important role in tumor growth and recurrence. These cells exhibit self-renewal and proliferation properties. CSCs also exhibit significant drug resistance compared with normal tumor cells. Finding new treatments that target CSCs could significantly enhance the effect of chemotherapy and improve patient survival. Notch signaling is known to regulate the development of the lungs by controlling the cell-fate determination of normal stem cells. In this study, we isolated CSCs from the human lung adenocarcinoma cell line A549. CD133 was used as a stem cell marker for fluorescence-activated cell sorting (FACS). We compared the expression of Notch signaling in both CD133+ and CD133- cells and blocked Notch signaling using the γ-secretase inhibitor DAPT (GSI-IX). The effect of combining GSI and cisplatin (CDDP) was also examined in these two types of cells. We observed that both CD133+ and CD133- cells proliferated at similar rates, but the cells exhibited distinctive differences in cell cycle progression. Few CD133+ cells were observed in the G2/M phase, and there were half as many cells in S phase compared with the CD133- cells. Furthermore, CD133+ cells exhibited significant resistance to chemotherapy when treated with CDDP. The expression of Notch signaling pathway members, such as Notch1, Notch2 and Hes1, was lower in CD133+ cells. GSI slightly inhibited the proliferation of both cell types and exhibited little effect on the cell cycle. The inhibitory effects of DPP on these two types of cells were enhanced when combined with GSI. Interestingly, this effect was especially significant in CD133+ cells, suggesting that Notch pathway blockade may be a useful CSC-targeted therapy in lung cancer.

Transareolar Video-Assisted Thoracoscopic Surgery in Females: A Novel Incision for Pulmonary Ground Glass Nodule Resection.

Purpose: Uniportal video-assisted thoracoscopic surgery (VATS) is recognized for its minimally invasive nature, widely adopted globally. However, the evident scarring it leaves often triggers psychological apprehension and resistance to surgery. Transareolar incision, known for its superior cosmetic outcome with no visible scars, poses challenges in women due to the risk of mammary gland damage. In this report, we present successful pulmonary ground glass nodule (GGN) resection using transareolar VATS in female patients, aiming to address these concerns. Materials and Methods: We retrospectively analyzed the clinical data of 35 female patients who underwent GGN resection through transareolar VATS between August 2020 and March 2022. Results: There were no serious complications or perioperative deaths in this cohort of 35 female patients undergoing GGN resection through transareolar VATS. The operations, including local resection or segmentectomy, had an average duration of 70.1 ± 26.4 minutes, with a tube duration of 4.7 ± 2.1 days and a hospitalization time of 7.2 ± 2.3 days. The surgical approach varied, with 21 cases using transareolar uniport, 8 cases assisted by a 3-mm tiny port, and 6 cases converted to two-port VATS. Scar outcomes varied, with 21 cases showing no scar, 8 cases displaying a microscar, and 6 cases presenting a dominant scar of 1.7 ± 0.5 cm. Postoperative pain scores at 1 week and 1 month were 1.9 ± 0.9 and 1.0 ± 0.9, respectively, and the wound numbness occurred in 2.86% (1/35) of cases. Regarding breast complications, 2 patients suffered delayed healing of the incision. No damage and inflammation of glands were detected by breast B-mode ultrasonography. Conclusions: The transareolar incision emerges as a novel approach for VATS in female patients, offering advantages in terms of pain management and cosmetic outcomes.

Comparison of gene mutation profile in different lung adenocarcinoma subtypes by targeted next-generation sequencing.

BACKGROUND: Disease prognosis after resection of lung cancer could be affected by pathological subtypes. In this study, we investigated the difference of gene variation and significantly altered pathways between adenocarcinoma in situ (AIS)/microinvasive adenocarcinoma (MIA) and invasive adenocarcinoma (IAC) subtypes to reveal the molecular mechanism of prognosis differences. METHODS: Sixty one tumor tissues were subjected to DNA extraction and customized 136 gene targeted next-generation sequencing. Comparisons between groups were performed with two-sided Fisher's exact test for categorical variables and two-tailed unpaired t test for numerical variables. RESULTS: A total of 402 somatic mutations involved in 70 genes were detected in all these samples, and 74.29% of these genes were mutated in at least two samples. PMS2, ARID1A, EGFR, and POLE were the most frequently mutated genes. ALK_EML4 fusion was observed in one IAC patient and RET_ KIF5B fusion in one AIS patient. A significant higher proportion of patients with TP53 gene mutation was observed in the IAC group (P = 0.0057). The average onset age in IAC group is 62.48 years, which is greater than other subtypes (P = 0.0166). It revealed that mutations in genes involved in the mTOR signaling pathway (56.52% vs 26.32%, P = 0.0288) and Hippo signaling pathway (34.78% vs 10.53%, P = 0.0427) were significantly enriched in IAC subtypes, suggesting the key involvement of mTOR and Hippo signaling pathways in lung tumor development and malignant progression. CONCLUSIONS: This study revealed the heterogeneity of gene mutations and significantly altered pathways between different lung cancer subtypes, suggesting the potential mechanism of different prognosis.

Two-Centimeter Single-Port Incision Minimizing Intercostal Nerve Damage Applied in Video-Assisted Thoracoscopic Surgery Segmentectomy for Pulmonary Ground Glass Nodules.

Purposes: We introduced a novel modified 2-cm single-port incision made by blunt separation minimizing intercostal muscle and nerve damage applied in video-assisted thoracoscopic surgery (VATS) segmentectomy, and compared it with the traditional single-port incision or the novel incision plus a 3-mm tiny port, aiming to explore a more minimally invasive single-port technique for VATS segmentectomy. Materials and Methods: We retrospectively analyzed the clinical data of 174 pulmonary ground glass nodule patients who received single-port VATS segmentectomy (54 modified 2-cm single port, 67 modified single port plus tiny port, and 53 traditional single port, respectively) in our medical center from May 2020 to December 2022. Three kinds of approaches were compared retrospectively, concerning their safety, feasibility, and postoperative pain. Results: There were no serious complications and mortality in either group. The blood loss, tube duration, and hospitalization time were comparable among the three groups (P > .05). The 2-cm single-port and 2-cm single-port plus tiny-port group were obviously more advantageous in the visual analog scores of postoperative pain, the wound numbness, incision healing and appearance than that in the traditional group (P < .05), while they were comparable. Notably, the operation time of the 2-cm plus tiny-port group was shorter than that of the 2-cm group (P < .05) and similar to the traditional single-port group. Conclusions: The 2-cm modified single-port applied for VATS segmentectomy is feasible and safe, and has obviously advantages in postoperative pain, numbness, and appearance of incision. With addition of tiny port, the convenience of the operation can be significantly increased without increasing pain. Our finding could provide a promising new incision mode for VATS segmentectomy.

Clinical significance of nutritional risk screening for older adult patients with COVID-19.

OBJECTIVES: The aim of this study was to assess the nutritional risks among older patients with COVID-19 and their associated clinical outcomes using four nutritional risk screening (NRS) tools: Nutrition Risk Screening 2002 (NRS 2002), Malnutrition Universal Screening Tool (MUST), Mini Nutrition Assessment Shortcut (MNA-sf), and Nutrition Risk Index (NRI). METHODS: We retrospectively analyzed the data of patients with COVID-19 older than 65 years who were treated in our hospital from January 28, 2020 to March 5, 2020, and explored the relationship between nutritional risk and clinical outcomes. RESULTS: A total of 141 patients with COVID-19 (46 common COVID-19, 73 severe COVID-19, and 22 extremely severe COVID-19) were enrolled in the study. NRS 2002 identified 85.8% of patients as having risk, with being identified 41.1% by MUST, 77.3% by MNA-sf, and 71.6% by NRI. The agreement strength was moderate between NRS 2002 and MNA-sf, NRI, fair between MUST and MNA-sf, NRI, fair between MNA-sf and NRI, poor between NRS 2002 and MUST (P < 0.01). After adjustment for confounding factors in multivariate regression analysis, patients in the risk group had significantly longer LOS, higher hospital expenses (except MNA-sf), poor appetite, heavier disease severity, and more weight change(kg) than normal patients by using NRS 2002, MNA-sf, and NRI(P < 0.05). CONCLUSIONS: The NRS 2002, MNA-sf, and NRI are useful and practical tools with respect to screening for patients with COVID-19 who are at nutritional risk, as well as in need of additional nutritional intervention.

Aß40 Promotes the Osteoblastic Differentiation of Aortic Valve Interstitial Cells through the RAGE Pathway.

Amyloid beta (Aß) peptide 40 enhances the activation of receptor for advanced glycation end products (RAGE) in immune-inflammatory diseases. RAGE exhibits several effects in the setting of numerous cardiovascular events. We hypothesized that the Aß40/RAGE pathway is involved in the osteoblastic differentiation of the valvular interstitial cell (VIC) phenotype, and RAGE knockout intervention could reduce the calcification of aortic valve interstitial cells (AVICs) by inhibiting the extracellular-regulated kinase1/2 (ERK1/2)/nuclear factor kappa-B (NF-κB) signaling pathway. To test this hypothesis, the activation of Aß40/RAGE pathway in human calcific AVs was evaluated with immunohistochemical staining. Cultured calcific VIC models were used in vitro. The VICs were stimulated using Aß40, with or without RAGE small interfering ribonucleic acid (siRNA), and ERK1/2 and NF-κB inhibitors for analysis. Our data revealed that Aß40 induced the ERK1/2/NF-κB signaling pathway and osteoblastic differentiation of AVICs via the RAGE pathway in vitro.

3,3',4',5,7-Pentamethylquercetin reduces angiotensin II-induced cardiac hypertrophy and apoptosis in rats.

Quercetin has been shown to possess beneficial pharmacological properties in treatment of heart disease, but lack of stability and bioavailability limits its clinical use. In this study, we investigated for the first time the effect of a methylated form of quercetin, 3,3',4',5,7-pentamethylquercetin (PMQ), on myocardial protection in rats. Angiotensin II was delivered to Sprague-Dawley rats subcutaneously, while PMQ (5 mg/kg) was administered by oral gavage; blood pressure was monitored daily. The production of NADPH oxidase was measured, and cardiac hypertrophy and apoptosis were detected. The results revealed that PMQ could downregulate the expression of the NADPH oxidase gene and reduce angiotensin II- induced cardiac hypertrophy and apoptosis in rats. Therefore, we believe that PMQ showed beneficial effects on myocardium in angiotensin II-administered rats, and its potential to be used for treatment of cardiovascular disease deserves further attention.

Fucosterol exerts antiproliferative effects on human lung cancer cells by inducing apoptosis, cell cycle arrest and targeting of Raf/MEK/ERK signalling pathway.

BACKGROUND: Phytochemicals have attained tremendous attention as the chemo-preventive and chemotheruptic agents. Fucosterol is a phytosterol that in prevalently found in marine algae and many other plant species. Previous studies have indicated the potential of fucosterol as an anticancer agent. However, the information on the anticancer activity of fucosterol against lung cancer as well as several other types of cancers is scantly. PURPOSE: The present study was designed to investigate the anticancer activity of fucosterol against a panel of lung cancer cell lines. METHODS: MTT and colony formation assays were used to determine the cell viability. DAPI and annexin V/PI staining assays were used for the detection of apoptosis. Cell cycle analysis was performed by flow cytometery. Boyden chamber assay was used to monitor cell migration and western blot analysis was used to determine the protein expression. In vivo evaluation was carried out in xenografted mice models. RESULTS: The results indicated that fucosterol inhibits the growth of the lung cancer cell lines. However, the anticancer effects were more profound against the A549 and SK-LU-1 cancer cells (IC50, 15 µM). In contrast, the anticancer effects of fucosterol on the non-cancerous lung cell lines were minimal. Further investigation revealed that the anticancer effects of fucosterol on the A549 and SK-LU-1 cells are due to the induction of apoptosis. Fucosterol significantly enhanced the expression of Bax and cleaved caspase-3 which was concomitant with decline in the expression of Bcl-2. Fucosterol also triggered G2/M cell cycle arrest of the A549 and SK-LU-1 cells which was associated with decrease in the expression of Cdc2, Cyclin A, Cyclin B1 and upregulation of the negative regulators of cell cycle progression (p21Cip1, and p27Kip1). Moreover, fucosterol could also inhibit the invasion of A549 and SK-LU-1 cells. Finally fucosterol could also inhibit the growth of xenografted tumours in mice. CONCLUSION: Taken together, fucosterol inhibits the growth of lung cancer cells and may prove to be a lead molecule for the treatment of lung cancer.

The completely mobilized remnant stomach: A new choice to reconstruct the esophagus in lower thoracic esophageal carcinoma with a history of distal gastrectomy.

OBJECTIVE: It is a significant surgical challenge to reconstruct esophagus for the patients following distal gastrectomy (DGE). Remnant stomach seems to be a better choice compared with colon or jejunal. But many complicated surgical methods were performed because of limitation of feeding vessels. We found the remnant stomach remained viable when all the feeding vessels were dissected. We used the completely mobilized stomach to reconstruct esophagus successfully in 29 lower thoracic esophageal carcinoma patients with a history of DGE. METHODS: The clinical data of 29 patients were retrospectively analyzed from August 2005 to March 2017 who accepted esophagoplasty by the completely mobilized remnant stomach. All the vessels of the remnant stomach were dissected including short gastric, posterior gastric, left gastric and left gastroepiploic vessels. The DGE included 2 Billroth I and 27 Billroth II. RESULT: No perioperative death, no remnant stomach necrosis occurred. One Leakage was the iatrogenic injury on the remnant stomach. The other postoperative complications were the pulmonary infection(5) and arrhythmia(4). CONCLUSION: The completely mobilized remnant stomach was viable and functional after dissecting all the feeding vessels. Application of it was a new and feasible surgical method to perform esophagoplasty with the simpler procedure and less complication.

Associations between autophagy, the ubiquitin-proteasome system and endoplasmic reticulum stress in hypoxia-deoxygenation or ischemia-reperfusion.

The activation of autophagy has been demonstrated to exert protective roles during hypoxia-reoxygenation (H/R)-induced brain injuries. This study aimed to investigate whether and how preconditioning with a proteasome inhibitor (MG-132), a proteasome promoter (Adriamycin, ADM), an autophagy inhibitor (3-methyladenine, 3-MA) and an autophagy promoter (Rapamycin, Rap) affected endoplasmic reticulum stress (ERS), the ubiquitin-proteasome system (UPS), autophagy, inflammation and apoptosis. Ubiquitin protein and 26S proteasome activity levels were decreased by MG-132 pretreatment but increased by ADM pretreatment at 2h, 4h and 6h following H/R treatment. MG-132 pretreatment led to the increased expression of autophagy-related genes, ER stress-associated genes and IκB but decreased the expression levels of NF-κB and caspase-3. ADM pretreatment led to the decreased expression of autophagy-related genes, ERS-associated genes and IκB but increased the expression of NF-κB and caspase-3. Pretreatment with 3-MA reduced the expression of autophagy-related genes, autophagy and UPS co-related genes, as well as apoptosis-related although the latter was increased by Rap pretreatment at 2h, 4h and 6h following H/R treatment. In vivo, pretreatment of rats with ADM, MG-132, 3-MA or Rap followed by ischemia-reperfusion (I/R) treatment resulted in similar changes. Proteasome inhibition preconditioning strengthened autophagy and ER stress but decreased apoptosis and inflammation. Autophagy promotion preconditioning exhibited similar changes. The combination of a proteasome inhibitor and an autophagy promoter might represent a new possible therapy to treat H/R or I/R injury-related diseases.

Autophagy decreases alveolar macrophage apoptosis by attenuating endoplasmic reticulum stress and oxidative stress.

To study the impact of autophagy on alveolar macrophage apoptosis and its mechanism in the early stages of hypoxia, we established a cell hypoxia-reoxygenation model and orthotopic left lung ischemia-reperfusion model. Rat alveolar macrophages stably expressing RFP-LC3 were treated with autophagy inhibitor (3-methyladenine, 3-MA) or autophagy promoter (rapamycin), followed by hypoxia-reoxygenation treatment 2 h, 4 h or 6 h later. Twenty Sprague-Dawley male rats were randomly divided into four different groups: no blocking of left lung hilum (model group), left lung hilum blocked for 1h with DMSO lavage (control group), left lung hilum blocked for 1 h with 100 ml/kg 3-MA (5 µmol/L) lavage (3-MA group), and left lung hilum blocked for 1 h with 100 ml/kg rapamycin (250 nmol/L) lavage (rapamycin group). Rapamycin decreased the unfolded protein response, which reduced endoplasmic reticulum stress-mediated apoptosis in the presence of oxygen deficiency. Rapamycin increased superoxide dismutase activities and decreased malondialdehyde levels, whereas 3-MA decreased superoxide dismutase activities and increased malondialdehyde levels. Thus, autophagy decreases alveolar macrophage apoptosis by attenuating endoplasmic reticulum stress and oxidative stress in the early stage of hypoxia in vitro and in vivo. This could represent a new approach to protecting against lung ischemia-reperfusion injury.