Heracleum paphlagonicum Czeczott Essential Oil Compound for Reducing Lung Inflammation in a CLP-Induced Sepsis Model

Abstract

This invention was developed to reduce the excessive inflammatory response and mitigate tissue damage seen in acute lung injury resulting from sepsis. Within the scope of this invention, it has been shown that doses of 100 mg / kg and 200 mg / kg of essential oil obtained from the seeds of Heracleum paphlagonicum Czeczott reduce the expression of pro-inflammatory mediators such as NF-κB, TNF-κB, IL-1β, IL-6, and HMGB1, and decrease cellular infiltration and structural damage in alveolar tissue.

Description

1 TARIFF CLP-Induced Sepsis Model: Reducing Lung Inflammation Heracleum paphlagon cum Czeczott Essential Oil Composition TECHNICAL AREA 5 This invention reduces lung inflammation in a CLP-induced sepsis model. heracleum paphlagon cum czeczott essential oil composition is pharmaceutical, It relates to the field of biotechnology and medical applications, especially those of plant origin. Essential oils are related to inflammation, oxidative stress, and immune response. It includes its use in the prevention and / or treatment of diseases. The invention encompasses these 10 within the scope of Heracleum paphlagon cum b tk snn seeds obtained Essential oil is used for systemic inflammatory conditions, especially sepsis, and these... Acute Lung Injury (ALI) and various pathological processes that develop depending on the situation. It includes its use in relation to its therapeutic effects. STATE OF THE ART Sepsis is the result of a severe host systemic inflammatory response to infection. The resulting heterogeneous clinical picture is that of multiple organs still present in intensive care units. Acute pulmonary insufficiency remains a significant problem. Injury (ALI) / acute respiratory distress syndrome (ARDS) is the most common sepsis and 20 It is an important complication (Dush anthan et al., 2011; Polat et al., 2013), Sepsis can easily occur in the early stages. Acute symptoms associated with sepsis. lung damage (ALI), edema, inflammatory cell infiltration, and consequently The deteriorated gas is characterized by its change in temperature. The complex pathophysiological mechanism of sepsis has led researchers to investigate sepsis 25 It has focused on researching pathogenesis and searching for effective treatment methods (Ap z (et al., 2025). Tumor necrosis factor (TNF)-α, IL-1 and IL-6 glycosides Initially, endotoxemia, fever, sepsis, and later autoimmune diseases. In studies on pathophysiology, therapeutic targets were identified and In clinical practice, cytokine-based treatment of inflammation became one of the cornerstones. 30 (Andersson & Tracey, 2011). However, new factors playing a role in the pathophysiology of sepsis. The search for stocks continues. Among these, the High Mobility group is box 1. (HMGB1) stimulates the immune system against sterile or infectious stimuli. 2 Thus, both innate and adaptive immune responses... Critical analysis plays a role in its regulation (Andersson & Tracey, 2011). Previous Studies have shown that HMGB-1 directly contributes to acute lung injury (ALI). has been determined (K m et al., 2005; Kwon et al., 2010; Ueno et al., 2004). New plant products or plant metabolites, new drugs and bioactive food additives 5 as an important source for substances (generally referred to as nutraceuticals) Its acceptance has become one of the most interesting topics in scientific research. (Bahador et al., 2016). Flowers with over 300 genera and over 3000 species. The Apaceae family is the most noteworthy in this regard, given its inclusion among the plants. Extracts obtained from various plants belonging to the family Ap aceae (Umbell ferae) 10 Cutting and anti-inflammatory effects have been determined (Dasht-Rahmatabad et al., 2007; Küpel et al., 2006). Ap aceae (Umbell ferae) family Heracleum species, fire various types of drugs such as decongestants, pain relievers, antiseptics, carminatives, and digestive aids. for purposes such as rheumatic infection, diarrhea, dysentery, back pain, and abdominal pain. It is widely preferred in traditional medicine due to its therapeutic properties. Research is being conducted on the essential oils of Heracleum species (Kose et al., 2020). Scientific studies on essential oils are increasing day by day. Essential oils are derived from plant parts. as, oct l acetate, oct l hexanoate, oct l octanoate, β-p nene, mr st sn, l monene and Hex l-butanoate is a complex mixture of compounds consisting mainly of gbb ioakt fb compounds. (Bahador et al., 2016; Ferrat et al., 2024; Hajhashem et al., 2009). 20 Resistance to antibiotics and other synthetic drugs used in sepsis treatment. and / or increased side effects by increasing antibacterial and antioxidant activity. Certain medicinal plants are suitable alternative treatments. Sepsis is an organ dysfunction associated with an irregular host response to infection. It is a heterogeneous clinical situation defined as follows: CLP model, immune system, 25 b no myasal and hemodynamic reactions similar to those observed in humans Therefore, it is represented by mimicking sepsis (Prescott & Angus, 2018; Starr (et al., 2014). Excessive and disordered inflammation plays a role in the pathogenesis of sepsis. the response is triggered particularly by activation of the nuclear factor kappa B (NF-κB) pathway. The release of toxins (TNF-α, IL-6, IL-1β and HMGB1) results in tissue damage and 30 It leads to organ failure. Current treatment approaches are mostly ineffective. It includes supportive treatment and broad-spectrum antibiotic applications, inflammatory process directly targeted specific pharmacological agents with effective efficacy 3 It has not yet been developed sufficiently. Currently, synthetic drugs have negative side effects. Concerns about possible side effects, natural products need to become more widely known, especially in medicine. There is an increasing demand for the use of plants in therapy (Mossa et al., (2013). Potential therapeutic properties of essential oils obtained from medicinal plant seeds. It has been reported that there are effects (Fatem et al., 2010; Fatem et al., 2018). 5 It can target the inflammatory response caused by sepsis, and suppress cytokine production. It can reduce tissue damage and also has a lower side effect profile. The development of new and effective therapeutic agents is a significant technical problem. This is what we are facing. Within the scope of this invention, the existing pharmacological framework in question... Innovative and effective treatment strategies in processes where treatments are insufficient. 10 It provides the opportunity for development. EXPLANATION OF THE INVENTION This invention explains the excessive inflammation seen in acute lung injury resulting from sepsis. It was developed to reduce the response and mitigate tissue damage. The invention Within this scope, 15 species of Heracleum paphlagon cum Czeczott were obtained from horse seeds. The 100 mg / kg and 200 mg / kg doses of the essential oil contained NF-κB, TNF-α, IL-1β, The expression of IL-6 and HMGB1, which are inflammatory mediators, was reduced. cellular infiltration and structural degradation in alveolar tissue are reduced It has been shown. This technical solution uses 20 naturally derived, biocompatible, and low-toxicity compounds. The use of fiber optic brachycardia (FBR) as an alternative approach in the treatment of lung damage caused by sepsis. It offers a solution where current pharmacological treatments are insufficient. Innovative and effective anti-inflammatory treatment strategies in inflammatory processes. It provides the opportunity for development. In this invention, pathological changes in lung damage caused by sepsis nn n 25 It has been determined that Heracleum paplan cum Czeczott is effective in alleviating it. Moreover, Heracleum paplan cum Czeczott essential oil has been shown to be effective against ALI, NF-KB, TNF-α, IL-6, and Reducing HMGB1 expression has been proven effective in preventing toxic storms. The essential oil used in the study was found to contain NF-κB, TNF-α, IL-1β, IL-6 and HMGB1. By regulating their expression, it suppresses the inflammatory response and the lungs 30 any technical solution or patent related to its tissue-protective effect It does not exist. Therefore, this invention differs from the existing art, Heracleum paphlagon cum Czeczott essential oil of sepsis-related lung disease 4 mitigating the damage, its potential effects revealed for the first time using molecular methods. It places.  Multifaceted Mechanism of Action: Heracleum paphlagonium oil, s tok n by suppressing the body's own gut microbiota storm (NF-kB, TNF-α, IL-6 and HMGB1) It prevents the damage caused by the immune response (ALI). 5  Dose-Dependent Efficacy: The study was conducted at specific doses such as 100 mg and 200 mg. By offering an optimized and effective treatment range, it provides a safe and effective treatment range. It defines it.  Natural and Plant Origin: Plant-based essential oils, compared to synthetic drugs. Because it is an oil, (in the appropriate formulation) the potential for side effects is less than 10%. low and more easily tolerated by the body It is anticipated.  Application variety: Pharmaceutical product, phytotherapeutic supplement or veterinary can be developed as a preparation Explanation of the Figures Figure 1. Representative photomicrographs of rat lung tissues. Figure 2a. NF-kB, TNF-α, IL-6, IL-1B in lung tissue caused by sepsis. Photographic images of HMGB1 expression. Figure 2b. NF-kB expression 20 Figure 2c. TNF-α expression. Figure 2d. IL-6 expression. Figure 2e. IL-1β expression. Figure 2f. HMGB1 expression. Figure 3. Components of Heracleum paphlagonum Czeczott Essential Oil 25 DETAILED DESCRIPTION OF THE INVENTION B tk material Collection of Specimens and Fieldwork Examples of B tk taxa were found within the borders of Kastamonu province, 30 km south of Tosya district center. Collected through field studies on the slopes and identified as part of the Flora of Turkey. This was done using the work titled "Flora of the Eastern Aegean Islands" (Dav s 1972). Araz In their studies, the fruits of the plants were preferred, lasting a total of 40 days over 2 years. As a result of the studies, 5000 grams of fruit samples were collected from each taxon. Drying of Samples After the collected plant materials, and especially the fruits, are brought to the laboratory... then, under environmental conditions, the diseased parts, soil and other foreign substances 5 It was cleaned by removing all substances and dried for approximately five days. B tk Examples nn Granulation The dried plant materials are processed to extract essential oils immediately before water distillation. To be granulated in order to be reduced to a size that can fit into the balloon of the distillation apparatus. This process was carried out using a grinding machine (28,000 rpm), and the samples were 15-10. The wind has been subjected to grinding processes. Obtaining Essential Oils from Plant Samples Extraction of essential oils from dried plant material, essential oil distillation equipment. The process was carried out and the resulting oils were stored at +4 °C for one week. In the water distillation method, 100 g of cleaned and ground fruit yields 15 The samples are placed in a glass balloon, and the connection between the balloon and the cooler is made of silicone. The oil was used to prepare the samples. The samples were boiled at 100 °C for 3 hours, then placed on a fingertip. The volatile oil and hydrolate phases of d-st-lat, which was condensed in the cooler, were separated. GC-MS Method (Volatile Compound Analysis) Chemical composition of Heracleum paphlagon cum Czeczott essential oil, gas 20 The analyses were performed using chromatography-mass spectrometry (GC-MS). The analyses, AOC-20 automatic sampling system equipped with Sh madzu GCMS-QP 2010 Ultra It was implemented in the system. Automatic sampler settings, solvent prewashing, six The solvent included the final wash and the sample wash. Both absorption and... The injection pulse speeds were set to high and the viscosity equalization time was 25. It was set to 0.2 seconds. The injection mode was set to normal. Gas Chromatograph separation, Rx-5MS capillary column (30.0 m length, 0.25 mm diameter, 0.25 This was performed using µm film thickness. The injector temperature was 250 °C. The settings were adjusted and injections were performed in a 5:1 split mode. Oven The temperature program was started at 40°C, held at this temperature for 3 minutes, and then... then the temperature was increased by 4 °C per minute to 240 °C and kept at this temperature for 10 minutes. 6 It was maintained throughout. Total oven program duration recorded was 63 minutes. Carrier gas. Helium (He) was used as the gas flow control. In pressure mode, the total flow rate is... The flow rate will be 13.7 mL / min, the column flow rate will be 1.78 mL / min, and the linear velocity will be 48.1 cm / s. It was operated as shown. The purging flow rate was set to 3.0 mL / min. Mass spectrometric detection was performed using electron ionization (EI) mode. 5 It was realized. The ion source temperature was reached 200 °C and the interface temperature was reached 250 °C. It's been adjusted. Solvent latency recorded as 3 minutes. MS data, 0.30 s scan time. and was obtained with a scanning speed of 1428 in scanning mode in the range of 40–450 m / z. The detector voltage was adjusted according to the automatic adjustment results of the device. Volatile Identification of components, comparison of obtained mass spectra with reference mass 10 This was done by comparing spectral libraries. The results are in Table 1. It has been summarized. animals In this study, W star alb no rats aged 12-13 weeks (weight: 200-300 gr) were used. 48 male Alb No W Star rats were used. The rats were fed a standard 12-hour feeding cycle. The light / dark cycle was maintained at temperatures between 19°C and 22°C. The rats were divided into 6 groups of 7 rats each; Group I (Sham): The cecum was mandible and purged but not ligated or perforated. Group II (CLP): Cecal Ligation and Perforation Model Group III (CLP-HPC50): Celiac Ligation and Perforation Model + Heracleum 20 paphlagon cum Czeczott EO 50mg / kg, Group IV (CLP-HPC 100): Cecal Ligation and Perforation Model + Heracleum paphlagon cum Czeczott EO 100mg / kg, Group V (CLP-HPC 200): Celiac Ligation and Perforation Model + Heracleum paphlagon cum Czeczott EO 200mg / kg, 25 Group VI (CLP-CEF100): Cebiliary ligation and perforation model + cefr axon 100 mg / kg. Rats were fasted for 12 hours before surgical intervention. Groups III, IV and 50 mg / kg, 100 mg / kg and 200 mg / kg Heracleum in 1 ml volume to single rats Paphlagon cum Czeczott's essential oil is used from 0 hours onwards for 30 days. Twelve hours later, intraperitoneal infusion ( .p.) was applied. The study was completed at 36 hours. terminated. 7 Creation of a CLP-derived sepsis model. Sepsis model will be induced by cecal ligation and puncture (CLP) method (M shra et al., 2008). Rats were administered Xylaz n (10 mg / kg) and ketam n (90 mg / kg) under anesthesia. The abdominal area is shaved and treated with R vanol, following asepsis and antisepsis rules. Accordingly, an incision 2 cm long was made on the left side of the medial line. 5 Through the incision, the muscles will be retracted to reach the peritoneal membrane, and the peritoneum will be... It opened and pulled the cecum out. Depending on the size of the cecum, use a 3 / 5 ratio at a distance of 3 mm. Using a 20G needle tip, the shape was determined. The apex of the cecum was shaped in a "+" direction. del nd .B rm ktar basmat le de klardan çir k dan sonra karnı ç back 10 It was closed by applying pressure to the skin, muscles, and dermis in sequence, leaving it as is. The rats, about to come out of anesthesia, will be taken to a safe area in cages. Sham In the group, an incision was made on the left side of the abdomen. The peritoneum, muscle, and skin were incised sequentially. The area was closed using the TDK (Turkish Language Association) procedure. 36 hours later, all animals were given general anesthesia. under the pretext of killing by overdose, whole blood samples were taken using intracardiac acid technique 15 It was removed. After removal, the lung tissue was divided into two pieces. : One piece - At 800°C, it was stored for western blot analysis, and the other samples were sent for histopathological and Immunoh stock was stored in 10% formalin solution for myasal analysis. Findings 20 F tok myasal Prof l of Heracleum paphlagon cum Czeczott Essential Oil The study also utilized Gas Chromatography and Mass Spectrometry (GC-MS) methodology. The production was achieved using Heracleum paphlagon cum Czeczott essential oil. The chemical composition was determined by GC-MS analysis, and a total of 48 volatile components were identified. is defined (Table 1). 25 8 Table 1. Volatile compounds of Heracleum paphlagon cum Czeczott. seeds detected by GC–MS method. components of oil 9 The main components of essential oil include Acetyl ester (39.97%) and Butyrate <hexyl->(16.82%) are included. The identified secondary components. between, Hexyl 2-Methyl Butyrate with 4.56%, Isobutyrate with 4.40% <hexyl->And Hexano c ac d, octyl ester stood out with 3.69%. H. cephalopathy findings 5 In the study, scoring results showed that lung damage was more prevalent in the CLP group. The index was significantly more severe according to the Sham group (Figure 1G, P<0.05). It was determined that lung damage in CLP-HPC groups was indexed as CLP-CEF100. Group F (Figure 2) showed no statistically significant difference (P>0.05). Lung Cellular infiltration in the intraalveolar septum, hyperemia and perbronchus-bronchus lymphoma d 10 The degree of hyperplasia in the tissue was evaluated. Statistical analysis findings are in Table. It is shown in Figure 1. Cell infiltration in the interalveolar septum CLP-HPC100 In the group (Figure 2D), the severity was statistically milder compared to the CLP group. It was noticed (Figure 2B, P<0.05). In the interalveolar septum, hyperemic and perbronchial- Bronchial lymph node hyperplasia in tissue, HPC groups and CLP-CEF100 groups 15 No statistically significant difference was found between them (Figure 2 CDE, P>0.05). Immunoh stock myasal Results In lung tissue caused by sepsis, NF-kB, TNF-α, IL-6, IL-1B, and Photographs of HMGB1 expression in Fig. 2a, statistical data in Table It was given in 2. 20 Table 2. Different groups of lung cancer, cytopathology and immunohistochemical stocks. Variation scoring. NF-kB expression is associated with intraalveolar septum cell NF-traction and perbronchial- NF-kB expression in bronchial lymphoid tissue, bronchial and bronchial epithelium cells, It was observed that the CLP group was more severe than the CLP-HPC200 group (Figure 3b). P<0.05). 5 TNF-α expression was higher in the CLP-HPC (100 and 200) and CLP-CE100 groups. In the CLP group, the severity of the condition was observed to be greater (P<0.01). Interalveolar TNF-α in septal cell infiltration and in bronchial and bronchial epithelium cells. CLP-HPC100 and CLP-CEF100 expression was higher in the sham and CLP groups. It was determined that the statistic was less severe in the group (Figure 10). 3c,P<0.01). IL-6 expression was more intense in the CLP group and in the CLP-HPC (100-200) group. It was observed that the severity was mild (P<0.01). Interalveolar septum cells IL-6 expression in nf treatment was higher in the CLP-HPC group compared to the CLP group (100-200) In the group, milder severity, perbronchial-bronchial lymphoid tissue CL-HPC100 15 According to the group, CLP-CEF100 group showed a lighter intensity of staining (P<0.001). Also, IL-6 expression in bronchial and bronchial leptomeningeal cells was observed in the CLP-HPC100 group. According to the group, the statistic was milder in severity (Figure 3d, P<0.05). IL-1B expression was higher in the CLP-HPC50 group compared to the CLP-HPC-200 group. It was noticed that it was more severe (P<0.01). Interalveolar septum cell 20 In the nf transduction group, IL-1B was used, and in the CLP-HPC200 group, CLP-HPC50 and CLP-HPC100 were used. It was determined that CLP-HPC 50 was expressed more mildly than the other group (P<0.01). Per bronchus-bronchus lymphoid tissue group: IL-1B, CLP-HPC200 and CLP-CEF100 It was observed that it was more severe than the other group (P<0.001). In addition, bronchial and bronchial lesions IL-1B expression in epthelium cellular matrix was higher in CLP-25 compared to Sham and CLP-HPC50 groups. Milder severity was observed in the HPC200 group (Figure 3e, P<0.001). HMGB1 expression was significantly higher in the CLP group, as were CLP-HPC50 and CLP- It was determined that the HPC100 group had mild severity (P<0.01). HMBG1, intra-veolar CLP- in septal cell infection and in bronchial and bronchial epithelium cells The HPC100 group had a milder immune reaction compared to the CLP group. was observed (Figure 1 3f, P<0.01).

Claims

11 REQUESTS 1. The invention relates to the prevention and / or treatment of sepsis-induced acute lung injury. It is an essential oil compound intended for use, and its properties are; Heracleum paphlagon cum Czeczott b tk sec , translated this b tk nn essential wind oil 5 The character of Çermes is damaged.

2. Claim 1 states that gbbr is an invention, and its characteristic is; Heracleum paphlagon cum Czeczott's essential oil is characterized by a concentration of 50-200 mg / kg.

3. Claim 1 states that gbbr is an invention and its characteristic is; Heracleum paphlagon cum Czeczott oil is preferably consumed in doses of 100 mg / kg or 200 mg / kg. character zed r.

4. Claim 1 states that gbbr is an invention, and its characteristic is; Heracleum paphlagon cum Czeczott essential oil contains HMGB1, NF-κB, TNF-α, IL-1β and IL-6 Pro-inflammatory mediators increase their expression simultaneously with NH2. etmes le karakter zed r. 15 25

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