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Qi-Long-Tian Formula Extract Alleviates Symptoms of Acute High

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Qi-Long-Tian Formula Extract Alleviates Symptoms of Acute High Fu et al. Respir Res (2021) 22:52 https://doi.org/10.1186/s12931-021-01645-8 RESEARCH Open Access Qi-Long-Tian formula extract alleviates symptoms of acute high-altitude diseases via suppressing the infammation responses in rat Xing Fu1, Chunyan Yang2, Bing Chen3, Kexing Zeng4, Siyuan Chen4 and Yi Fu5* Abstract Background: Chinese Yunnan Province, located in the Yunnan–Guizhou Plateau, is a famous tourist paradise where acute high-altitude illness common occurs among lowland people visitors due to non-acclimatization to the acute hypobaric hypoxia (AHH) conditions. Traditional Chinese medicine, such as Qi-Long-Tian (QLT) formula, has shown efectiveness and safety in the treatment of acute high-altitude diseases. The aim of this study was to clarify the thera- peutic mechanisms of this traditional formula using a rat model in a simulated plateau environment. Methods: Following testing, lung tissue samples were evaluated by hematoxylin–eosin staining and for biochemical characteristics. mRNA-Seq was used to compare diferentially expressed genes in control rats, and in rats exposed to AHH and AHH with QLT treatment. Results: Infammation-related efectors induced following QLT treatment for AHH included MMP9 and TIMP1, and involved several phosphorylation signaling pathways implicated in AHH pathogenesis such as PI3K/AKT and MAPK signaling. Conclusion: This study provides insights into the major signaling pathways induced by AHH and in the protective mechanisms involved in QLT formula activity. Keywords: Acute hypobaric hypoxia, Qi-Long-Tian, mRNA-seq, Acute high-altitude diseases, Infammation Background PO2 drops by 1 mmHg. Millions of individuals, includ- Te northern part of Yunnan Province, China is located ing the military, mountain rescuers, and mountaineers, on a plateau with an average altitude of over 2000 m. With currently work or live at high altitudes, and are exposed increasing altitude, the atmospheric pressure decreases to the risk of mountain sickness. Physiological studies exponentially, resulting in a progressive reduction in the have shown that the altitude suitable for human sur- ambient partial pressure of oxygen (PO 2), termed hypo- vival is between 500 and 2000 m [1]. Individuals experi- baric hypoxia. When the altitude increases by 100 m, the ence difculty breathing and altitude symptoms appear atmospheric pressure usually drops by 5 mmHg, and the above 2000 m, while saliva secretion begins to decrease at 3000 m. Most individuals experience a state of incapaci- tation at 6000 m and loss of consciousness and hallucina- *Correspondence: [email protected] tions occur at 7000 m [2]. 5 The Third Afliated Hospital of Yunnan University of Chinese Medicine, Kunming Municipal Hospital of Traditional Chinese Medicine, No. 2628 When lowland people visit plateaus, a syndrome Xiangyuan Street, Chenggong District, Kunming 650500, Yunnan, China caused by poor adaptability to the acute hypobaric Full list of author information is available at the end of the article hypoxia (AHH) environment called acute mountain © The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creat iveco mmons .org/licen ses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creat iveco mmons .org/publi cdoma in/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Fu et al. Respir Res (2021) 22:52 Page 2 of 12 sickness (AMS) may occur [3]. Te principal symptoms with hypoxia pulmonary hypertension [17, 18], but the of AMS include headache, nausea, vomiting, fatigue, underlying mechanisms are still unclear. dizziness, and sleep disturbance [4]. Although AMS In the present study, we investigated the mRNA profle has been recognized over the past two centuries, little of lung tissues from rat models exposed to conditions of is known about the fundamental causes of these symp- normoxia, AHH, or to AHH with Qi-Long-Tian treat- toms [5]. Yunnan Province is a tourist paradise, where ment. Te results showed that several infammation- the number of tourists exceeded 600 million in 2020. related mediators were signifcantly induced under AHH It is foreseeable that AMS would seriously threaten the conditions, which also responded to QLT treatment. We health of travelers visiting the plateau. deduce that these molecule candidates may mediate the A series of physiological responses occur to adapt action of QLT in vivo. Tis study provides deeper insight to AHH conditions. In some cases, these maladaptive into the mechanisms of action of QLT in the treatment of responses may predispose afected individuals to vari- AHH-related diseases. ous forms of high-altitude illness. Te lungs are the frst organ to sense changes in barometric pressure Methods Preparation of the extract of QLT formula and the PO2 in the environment and are closely asso- ciated with the body’s maladaptive responses to AHH Te composition of the QLT formula includes Rhodiola [6], which result in signifcant injuries caused by hypo- sachalinensis, Panax notoginseng, and dried Pheretima baric hypoxia, such as high-altitude pulmonary edema asiatica at a ratio of 10:5:2. Te raw medicinal materials (HAPE). were obtained and prepared by the Kunming Municipal HAPE is noncardiogenic pulmonary edema that may Hospital of TCMs. Te materials were diluted 10 times occur in unacclimatized persons within 2 to 4 days of in distilled water (w/v) and boiled twice for 1 h each time ascent to altitudes above 2500 m [7]. Clinical fndings under continuous stirring. After fltration, the residue include tachycardia, tachypnea, low-grade fever, hypox- was boiled with 8 volumes of purifed water for another emia, and unilateral or bilateral inspiratory crackles [8]. 2 h and fltered. Tereafter, the combined fltrates were Risk factors for HAPE include a prior history of the dis- subjected to centrifugation at 1500×g and the superna- order, male sex, rapid ascent, higher altitudes, pre-exist- tants were concentrated under vacuum to a fnal concen- ing respiratory infection, and intense exercise [9–11]. tration of 2 g/mL and stored at − 80 °C until use. Susceptible individuals develop exaggerated hypoxic pul- monary vasoconstriction and may experience a large rise Animals in pulmonary artery pressure when exposed to hypoxia. Specifc pathogen-free male Wistar rats (230 ± 20 g; As these changes are distributed unevenly within the pul- 6-weeks-old) were obtained from the Laboratory Ani- monary vascular bed, regional over perfusion of capil- mal Center of Kunming Medical University. All rats were laries occurs, leading to a ‘stress failure’ of the blood-gas maintained in an environment at 23 ± 2 °C with 50 ± 5% barrier, increased permeability, and pulmonary edema humidity and a 12 h light–dark cycle. Water and food [12]. Tis noninfammatory process may be accelerated were provided to animals ad libitum. Te experiments by impaired alveolar fuid clearance [13]. started after a week of adaptation. Te experimental pro- Traditional Chinese medicine (TCM), which has long tocol was approved by the Experimental Animal Care been practiced in the clinic in Asian countries, is an and Ethics Committees of Kunming Municipal Hos- important choice for preventing and treating disease. pital of Traditional Chinese Medicine (Approval No.: Formulae based on TCM theories are reported to be R-082018112). efective in treating hypoxia pulmonary hypertension which is a symptom of HAPE at the early stage. Te Qi- Rat model preparation and administration Long-Tian (QLT) formula is composed of three herbs Te 130 rats were randomized and divided into the blank (Rhodiola, Panax notoginseng, and dried pheretima at a (I), AHH (II), and AHH with QLT treatment (III) groups. ratio of 10:5:2) [14]. As a component of QLT, Rhodiola Animals of the blank group (n = 10) remained at sea-level has been used in the clinic as a lung protecting agent atmospheric pressure within the same laboratory condi- and can improve cardiopulmonary function, alleviate tions. Animals of groups II and III (60 rats for each) were pulmonary hypertension, and reduce the probability of exposed to AHH for a total of 24, 48, or 72 h (20 rats for the occurrence of HAPE [15, 16]. A previous study has each duration) in a decompression chamber (Feng Lei, shown that this formula reduced endothelin-1, vascular Guizhou, China) at a simulated altitude of 6000 m. Te endothelial growth factor, interleukin (IL)-1β, and tumor animals of the hypobaric hypoxia group III received 8 g/ necrosis factor (TNF)-α levels, and upregulated nitric kg (20 rats for each duration) QLT by gavage with once a oxide synthase (NOS) expression in experimental rats day for the duration of the experiment. Te rats of group Fu et al. Respir Res (2021) 22:52 Page 3 of 12 II received the same volume of saline intragastrically. Te at RT. Chloroform (0.2 mL) was added into an EP tube airfow of the hypobaric hypoxia decompression chamber and mixed vigorously for 30 s, and then allowed to sit was maintained 10 L/min during exposure to prevent the for another 5 min at RT. Next, the lysate was centrifuged accumulation of exhaled gases.
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