Therapeutic Modulation of Ischemic Preconditioning by Alantolactone in Diseased Rat Hearts

INTRODUCTION:

Hyperlipidemia is a major contributor to ischemic heart disease (IHD). Ischemia-reperfusion (IR) injury describes the damage that tissue sustains when blood flow is re-established after a period of ischemia [1,2]. Extended periods of ischemia can cause irreversible cell damage and lead to apoptosis. Ischemic heart diseases are associated with oxidative stress and an excess of intracellular calcium [3,4]. Oxidative stress as well as calcium overload led to myocardial cell dysfunction and lead to apoptosis, which markedly impair function of cardiac [4,5]. The phenomenon of Ischemic preconditioning (IPC) offers noteworthy protection in contrast to subsequent episodes of IR injury. Protective mechanisms such as protein kinase C (PKC) activation and mitochondrial ATP-sensitive potassium (K_ATP) channel opening are triggered during the first phase. These processes aid in reducing cell damage and improving cell survival [6,7].

Ischemic preconditioning (IPC) is an internal mechanism to safeguard the heart from ischemia. During IPC, the myocardium experiences a short but severe period of sublethal ischemia and reperfusion [8,9] I/R-induced damage is linked to endothelial dysfunction, oxidative stress, and the inhibition of apoptosis-necrosis, and research has shown that IPC can mitigate these effects [10] Activation of the PI3K-Akt pathway and inhibiting the MPTP opening lead to development of mitochondrial ATP-sensitive potassium (KATP) channels [11,12].

Alantolactone, compound isolated from the medicinal herb Inula racemosa found in regions of Himalayas. [13,14]. Alantolactone, an active chemical constituent present in plant Inula Racemosa, exhibits property against cancer and inflammation, also possess anti-bacterial, and anti-fungal properties [15]. Solvent extraction and column chromatography have traditionally extracted and purified AL (Stojakowska et al., 2006; Ma et al., 2013), but these methods are time-consuming, wasteful, toxic, and unsuitable for large-scale production. We require efficient methods to extract alantolactone compounds from I. racemosa. Reduced operating temperature and pressure, reduced extraction time, compatibility with the environment, goods selectivity, a single step from extraction to separation, and the absence of residual solvents are all advantages of supercritical fluid extraction (SFE) over traditional solvent extraction methods. Among SFE solvents, supercritical CO2 is the most widely used. It can be extracted using low temperatures and pressures because it is a non-toxic, inert substance. The diffusion coefficient, selectivity, and solvent separation may all be improved by adjusting the working parameters (Demirbas¸, 2001; Schacht et al., 2008; Kruse and Vogel, 2010).

It may protect the heart against myocardial damage brought on by ISO by lowering oxidative stress and controlling ventricular and haemodynamic processes [16] In hyperlipidemia (HL), ischemic preconditioning (IPC) is less effective at inhibiting MPTP opening [17]. MPTP is a protein complex found in the inner mitochondrial membrane that can become permeable in certain pathological circumstances, such as stroke, traumatic brain damage, and ischemia reperfusion injury [18]. However, it is unknown whether combining IPC with alantolactone has any cardioprotective effects in HL condition and MPTP plays major role.

II. MATERIAL AND METHOD:

The study utilized Wistar rats, each weighing between 200 and 250 grams.

1. Investigational Chemicals and drugs

Procured chemicals and reagents were used in this investigation was of analytical quality and was made fresh and used immediately. The supplier of alantolactone is Sigma-Aldrich Chemicals in Bangalore, India. All study methods were authorised by the Institutional Animal Ethics Committee (KNIMT / PHAR / IAEC/ 18/ 05) in accordance with national regulations on experiments with animals.

2. Development of Hyperlipidaemic Condition

To induce experimental hyperlipidemia, a specially formulated high-fat diet was administered for six weeks. The diet consisted of specific proportions of corn starch, sugar, butter, casein, fiber and other nutrients. This established diet regimen effectively induced hyperlipidemia, as confirmed by measuring serum cholesterol and triglycerides using Span Diagnostics' assay kit.[14,19]

3. Rat Heart Isolation and Mounting for Physiological Studies

Heparin 500 IU/L, IP was used as anticoagulant and sodium pentobarbital (60 mg/kg, IM) was used to anaesthetise wistar rats. Following their removal, the hearts were placed on a Langendorff apparatus and retrogradely perfused with oxygenated KH buffer.

4. Experimental design for IPC studies in diseased hearts

Nine groups, each with six rats, participated in the study. All animals were acclimatized for 7 days before being randomly assigned to groups. Figure 2 shows a diagrammatic representation of the experiment protocol.

Group 1- Sham Control Group: After the mounting the isolated heart on lagendroff’s apparatus and 10 minutes of stabilisation, is subjected to 190 minutes with KH buffer. There is no involvement of global ischemia.

Group 2- IR (Ischemia-reperfusion) Control Group: Heart on lagendroff’s apparatus, after 10 minutes of stabilisation, exposed to global ischaemia for thirty minutes and reperfusion for one hundred and twenty minutes.

Group 3 - IPC (Ischemic Preconditioning) Control: The mounted heart is undergone with the four cycles of ischaemia for five minutes, reperfusion for five minutes, and then 30 minutes of global ischaemia followed by reperfusion for 120 minutes.

Group 4 – IR+HL (Hyperlipidaemic rats): The hyperlipidaemic rat heart after 10 minutes of stabilisation subjected with Ischemia reperfusion cycle.

Group 5- IPC +HL: After ten minutes of stabilisation, the isolated hearts of hyperlipidaemic rats underwent IPC cycle.

Group 6- AL (Alantolactone) +IR + HL: Before causing 30 minutes of global ischaemia and 120 minutes of reperfusion, the separated hearts from hyperlipidaemic rats were pre-treated with AL (50 mg/kg, given intraperitoneally) 30 minutes beforehand.

Group 7- IPC+AL+HL: Alantolactone (50 mg/kg, given intraperitoneally) was pre-treated 30 minutes prior to ischemia for 30 minutes and reperfusion for 120 minutes, and hearts from hyperlipidaemic rats were subjected to the IPC protocol, which consists of four cycles of 5 minutes of each ischaemia and reperfusion.

Group 8- IPC + Atr (Atractyloside) + HL: Four cycle if IPC is subjected to the heart and in the last cycle. Atractyloside was re-perfused followed by 30 minutes of ischemia and reperfusion for 120 minutes.

Group 9- IPC + AL+ Atr + HL: A treatment comprising atractyloside (Atr), alantolactone (AL) (50mg/kg), and ischaemic preconditioning (IPC) was administered to hyperlipidaemic rats. Four preconditioning sessions were given and Atractyloside was introduced during the final ischaemic preconditioning phase. After that, the hearts were given to ischaemia 30 minutes and reperfusion 120 minutes.

5. Evaluation of Myocardial Injury

LDH level and CK-MB level in the coronary effluent were measured using a commercial diagnostic kit from Coral Clinical System, Goa, India, in order to determine myocardial damage. International units per litre (IU/L) are used to display the findings[12,20].

6. Myocardial Infarct Size Assessment

From the Langendorff apparatus, the heart was extracted. The ventricles were kept at -4°C for the night after the auricles and the aortic root were removed. After that, the frozen ventricles were divided into homogeneous pieces that were 1-2 mm thick. After soaking for 30 minutes at 37°C in a 0.2 M Tris buffer solution (pH 7.4) containing a 1% w/v solution of triphenyl tetrazolium chloride (TTC), the slices are placed in an incubator. The infarcted cell region was left unstained, but viable cells were stained a deep red, due to the conversion of TTC to red formazan pigment by NADH and dehydrogenase enzymes. The infarct size was measured using the volume approach [14,21].

7. Myocardial Reduced Glutathione (GSH)

Mixtures of 0.1 ml supernatant, 2.0 ml phosphate buffer (0.3 M, pH 8.4), 0.4 ml double-distilled water, and 0.5 ml DTNB (5,5 dithiobis-2-nitrobenzoic acid) were used to assay GSH. After 10 minutes of incubation, the absorbance at 412 nm was measured for the combination. Microgrammes per millilitre of wet weight is the unit of measurement used to express the results [21].

8. Superoxide Dismutase (SOD)

To make a total volume of 0.6 mL, combine 0.1 mL of 186 µM phenazine methosulphate, 2.3 mL of 300 mM nitro blue tetrazolium, and 0.2 mL of 780 µM NADH with sodium pyrophosphate buffer (pH 8.3). The combination was left to incubate at 30°C for 90 seconds. To stop the reaction, one millilitre of acetic acid was added. After adding 4 mL of n-butanol, the mixture was spun in a centrifuge at 3000 rpm for 10 minutes (see Table 1)[14].

9. Estimation of Catalase

The material was placed into a 3 ml cuvette along with 1.95 ml of 50 mM phosphate buffer (pH 7.0). We assessed the changes in absorbance at 240 nm at 15-second intervals for 30 seconds after administering 1 ml of 30 mM hydrogen peroxide. A unit per milligramme of protein represents the results [14]

10. Investigation of Mitochondrial Efficiency and Resilience

a. Separation of Mitochondria from Cardiac Tissue

Following perfusion in the control group, mitochondria were quickly extracted from the heart in each experimental group using a conventional procedure. Following reperfusion, mitochondria were extracted from the heart for the remaining groups, and they were then exposed to the HL models [22]

b. Assessment of Mitochondrial Function

To quantify mitochondrial activity, formazan, the reduced form of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), was generated. Spectrophotometry was used to measure the quantity of formazan produced at 595 nm. The results were displayed as the rate of formazan synthesis in milligrams per minute per milligram me of protein [14,23]

11. Immunoblot Assay

To prepare the heart tissue for protein extraction, it was quickly frozen in liquid nitrogen and kept at -80°C. A supernatant containing soluble proteins was obtained for further analysis by forcefully disrupting the thawed tissue and then centrifuging it for 20 minutes. The amount of protein in every sample was determined by use of a modified Bradford technique. Gel electrophoresis was performed by loading 35 µg of the isolated proteins onto SDS-PAGE gels and then separating them based on molecular weight. After proteins were deposited onto PVDF membranes, primary antibodies that target particular proteins were treated with the membranes. The next step was to apply secondary antibodies diluted at 1:1000. After that, the membranes were left to incubate at room temperature in the dark for one hour. β-actin was used as a standard to ensure uniform loading in all samples. An E-Gel Imager (Tanon 5200 Multi, Shanghai, China) was used to image bands and an enhanced chemiluminescence test was used to visualise bound antibody complexes [20].

12. Analysis of Histopathology and the Content of Cardiac Collagen

In a 10% buffered neutral formalin solution, the heart tissue was preserved. Following fixation, the tissues were fixed in paraffin wax, and a microtome was used to cut transverse mid-ventricular slices that were about 5 µm thick. Haematoxylin and eosin staining was then applied to the slices in order to assess myocyte size and necrosis. Additionally, the collagen content was examined using Picrosirius Red F3BA staining to evaluate fibrosis [21,24]

13. Statistical Analysis

Mean ± S.E.M. data were provided for each group of six animals. In order to analyse the data statistically, one-way ANOVA and Tukey's multiple comparison test were employed. We considered p-values below 0.05 to be statistically significant.

RESULTS:

1.High-Fat Diet effect on Body Weight of rats

Rats fed a hyperlipidaemic diet for six weeks had a significant rise in body weight. Nevertheless, the alantolactone-treated hyperlipidaemic rats' body weight did not decrease (Figure 3).

2. Hyperlipidaemic Diet effect on Serum Cholesterol and Triglyceride Levels

Rats given a hyperlipidaemic diet for six weeks showed a considerable rise in triglyceride as well as total cholesterol in their blood. The hyperlipidaemic rat’s high levels of triglycerides as well as total cholesterol were not brought down by alantolactone therapy (Figure 4).

3. IPC effect on the LDH and CK-MB release in Control and High-Fat Diet Groups

Both normal and hyperlipidaemic rat hearts released considerably more LDH and CK-MB after a 30-minute global ischaemia and a 120-minute reperfusion. Treatment with alantolactone had no additional effect on the LDH and CK-MB release in IR control HL rat hearts. In normal rat hearts but not in hyperlipidaemic ones, ischaemic preconditioning markedly reduced the LDH and CK-MB release caused by ischemia-reperfusion. However, when Alantolactone was administered to hyperlipidaemic rat hearts, this decrease was markedly recovered. In hyperlipidaemic rats treated with Alantolactone, the drop in LDH and CK-MB brought about by Alantolactone administration was substantially counteracted by giving atractyloside during the last reperfusion episode of ischaemic preconditioning (Figures 5a and 5b).

4. IPC Effect on Ischemia and Reperfusion caused Myocardial Infarction in Control and High-Fat Diet Groups

Both normal as well as hyperlipidaemic rat hearts showed a substantial growth in size of myocardial infarct after a 30-minute global ischaemia and a 120-minute reperfusion interval. In ischemia-reperfusion control hyperlipidaemic rat hearts, alantolactone had no effect on the size of myocardial infarcts. Ischaemic preconditioning considerably decreased the extent of myocardial infarcts caused by ischemia-reperfusion in normal rat hearts, but not in hyperlipidaemic ones. In hyperlipidaemic rats treated with Alantolactone, the infarct size reduction caused by Alantolactone therapy was significantly eliminated when atractyloside was administered during the last reperfusion episode of ischaemic preconditioning (Figure 6).

5. IPC Effect on Oxidative Stress Markers in Control and High-Fat Diet Groups

In both normal and hyperlipidaemic rat hearts, endogenous antioxidant levels (GSH, SOD, and catalase) were markedly reduced by a 30-minute global ischaemia followed by a 120-minute reperfusion. The ischemia-reperfusion control hyperlipidaemic rat hearts' GSH, SOD, and catalase levels were unaffected by the alantolactone therapy. In normal rat hearts but not in hyperlipidaemic rat hearts, ischaemic preconditioning considerably reduced the ischemia-reperfusion-induced drop in GSH, SOD, and catalase levels. However, in hyperlipidaemic rat hearts treated with Alantolactone, this mitigation brought on by ischaemic preconditioning was considerably recovered. The restoration of GSH, SOD, and catalase levels brought on by Alantolactone therapy was significantly reversed in hyperlipidaemic rats treated with Alantolactone when atractyloside was administered during the last reperfusion event of ischaemic preconditioning (Table 1).

6. IPC Effect on mitochondrial function and integrity in Control and High-Fat Diet Groups

According to formazan levels and TMRM fluorescence intensity (which indicates mitochondrial integrity), alantolactone (AL) affects the alterations in mitochondrial function brought on by ischaemic preconditioning (IPC). Post hoc analyses showed that hyperlipidaemic (HL) rat hearts had considerably worse mitochondrial function than the sham control. AL enhanced IPC's decreased mitochondrial activity in HL hearts compared to controls as well as IPC+HL groups. Nevertheless, when both atractyloside and AL were given, the improvement in mitochondrial function that IPC had brought about in HL rat hearts was lessened (Figure 7a and 7b).

7. IPC Effect on Histopathological Analysis Control and High-Fat Diet Groups

There is no discernible cardiac damage in the sham control group's photomicrographs, which display normal heart muscle architecture (endocardium, epicardium, papillary muscles, and vasculature). Photomicrographs of the I/R control treatment group show crucial myonecrosis, myophagocytosis, and lymphocytic infiltration. Furthermore, the sub-endocardium has notable chronic inflammation. While myonecrosis and sub-endocardial alterations persist, the IPC-treated group exhibits less inflammation and myonecrosis. Although there is more myonecrosis in the IPC + HL group than in the IPC group, there is less inflammation and myo-phagocytosis, and subendocardial vacuolar alterations are observable under a microscope.

Under the microscopical examination, the IPC + AL group exhibits less myo-necrosis and decreased inflammation. Less severe myonecrosis, myo-phagocytosis, and lymph vascular infiltration are seen in the IPC + HL + AL treated group, but oedema and inflammatory cell infiltration are less common. In contrast to the IPC and IPC + HL + AL groups, myonecrosis, inflammation, and decreased myo-phagocytosis are shown in the IPC + HL + AL + Atr treated group (Figure 8).

8. AL Facilitates IPC-Induced Attenuation of PI3K Expression in HL Rat Heart

Figure 9, illustrates how alantolactone (AL) dramatically alters IPC-induced alterations in PI3K expression in hyperlipidaemic (HL) rat hearts. PI3K expression is much higher in AL-treated rat cardiac tissue than in the sham control group, according to post hoc test results. In rat hearts treated with AL, ischaemic preconditioning (IPC) decreases PI3K expression. In HL rat hearts, AL therapy intensifies the IPC-induced decrease in PI3K expression. Furthermore, IPC-mediated alterations in PI3K expression in HL rat hearts are considerably impacted by the combination of atractyloside and AL.

: GSH, Myocardial Reduced Glutathione; SOD, Superoxide Dismutase; CAT, Catalase.

Table:1

Figure s

Figure:1

Figure: 2

Figure: 3

Figure: 4

Figure: 5

Figure: 6

Figure: 7

Figure: 8

Figure: 9

Legend for Table

Table : 1 All values are mean ± S.D. (n=6). The following comparisons were made: ^aP<0.05 with sham control, ^bP<0.05 with I/R control, ^cP<0.05 with IR control, ^dP<0.05 with IPC control, ^eP<0.05 with IR control, ^b,d,fP <0.05 with IPC control and IPC + HL, ^f,gP <0.05 with IPC control and IPC +AL +HL, ^f,hP <0.05 with IPC control and IPC + AL + HL.

Legends of Figure

Figure: 1 Structure of Alantolactone

Figure:2 The schematics representation of the experimental methodology for various groups of animals.

Figure :3 Effect of High-Fat Diet on Body Weight

Figure: 4 Serum Cholesterol and Triglyceride

Figure: 5 Alantolactone restored IPC-mediated alterations in LDH and CKMB in the hearts of HL rats respectively. All data are shown as mean standard deviation (n=6) unless otherwise specified. It was discovered that I/R control was more successful than sham in terms of ^aP, and that it was more effective than I/R control and I/R + HL in terms of ^cP and ^dP. Although IPC control and IPC + HL were shown to be more successful in terms of objective evidence, IPC control was found to be more effective in terms of anecdotal evidence. Using a one-way ANOVA and the Bon-ferroni Post hoc test, it was discovered that ^{e, h, ,j}P0.05 was substantially lower than that of IPC control and IPC + AL + HL.

Figure:6 The HL rat heart's infarct was altered by IPC, but AL was repaired. For all values, mean ± S.D. (n=6) is used. The following comparisons were made: ^aP<0.05 versus sham control, ^bP<0.05 versus I/R control, ^cP<0.05 versus IR control, ^c,dP<0.05 versus I/R control and I/R +HL, ^eP <0.05 versus IPC control, ^e,fP <0.05 versus IPC control and IPC + HL, and ^e,gP <0.05 versus IPC control and IPC +HL. (one way ANOVA followed by Bon-ferroni Post hoc test) ^e,hP < 0.05 compared with IPC control and IPC + HL, ^jP < 0.05 compared with IPC control and IPC + AL + HL.

Figure:7a and 7b. In the HL rat heart, AL corrected in IPC induced alterations in function (formazan production) and mitochondrial integrity (TMRM fluorescence intensity) respectively. For all values, mean ± S.D. (n=6) is used. ^aP<0.05 against sham control, ^bP<0.05 against IR control, ^cP<0.05 against IR control, ^c,dP<0.05 against IR control and IR +HL. ^eP <0.05 against IR control. ^e,fP <0.05 against IPC control and IPC + HL. ^e,gP <0.05 against IPC control and IPC +HL. ^e,hP <0.05 against IPC control and IPC + Al + HL (one way ANOVA followed by Bon-ferroni Post hoc test).

Figure: 8 Heart histological alterations brought on by IPC include normal and hyperlipidaemia. The impact of hyperlipidaemia on the heart in patients with pretreatment AL, AtR, or pretreated AL plus Atr, as reported by histopathology. Whereas sham control displays normal myocardial cell architecture, I/R control displays necrotic alterations in myocardial tissue. IPC control indicates that myocardial tissue has undergone a regeneration phase. Myocardial tissues exhibit necrotic changes when I/R + HL, necrotic changes when I/R + AL + HL, less regenerative changes when IPC + HL, more regenerative changes when IPC + AL + HL, and necrotic changes when IPC + AL+ Atr + HL.

Figure: 9 Changes in the level of expression in hyperlipidaemic rat heart tissue were mediated by AL restored in IPC. The arbitrary unit of PI3K expression to β-actin is used to express the PI3K protein histogram. For all values, mean ± S.D. (n=6) is used. The Bon-ferroni Post hoc test was used to compare the following: ^aP<0.05 with sham control, ^bP<0.05 with IR control, ^cP<0.05 with IR control, ^c,dP<0.05 with IR control and IR +HL, ^b,eP <0.05 with IR control and IPC + HL, and ^fP<0.05 with IPC control and IPC + AL + HL.

DISCUSSION:

Specifically, the results show that alantolactone (AL) significantly improves the cardioprotective effects of ischaemic preconditioning (IPC) in hearts with hyperlipidaemia (HL), mostly via reducing the opening of the mitochondrial permeability transition pore (MPTP). The development of hyperlipidaemia was confirmed by the considerable increases in body weight, serum cholesterol, and triglyceride levels in rats that were fed a high-fat diet for six weeks. Although AL treatment did not normalize these elevated levels, the findings indicate that dietary interventions alone are inadequate to address these risk factors. This underscores the necessity for complementary treatments.

IPC significantly reduced myocardial infarction volume size in normal rat hearts, but this cardioprotection was lessened in HL rat hearts. Alantolactone (AL) treatment did not independently decrease infarction volume size in HL rats infer with ischemia-reperfusion (I/R) injury. However, adding atractyloside (Atr) during the final reperfusion episode nullified the IPC-induced reduction in infarct size in AL-treated HL rat hearts. This suggests that the effectiveness of IPC in reducing myocardial damage is compromised in the presence of hyperlipidemia, and AL's protective effects are influenced by the presence of Atr.

The biomarkers CK-MB and LDH, indicators of myocardial injury, were significantly elevated after I/R injury in both normal and hyperlipidaemic (HL) rat hearts. While IPC reduced these markers in normal hearts, it did not have the same effect in HL hearts. The IPC-induced decrease in CK-MB and LDH levels in HL hearts was reversed by alantolactone (AL) therapy, indicating that AL may mitigate some of the detrimental effects of hyperlipidaemia on myocardial damage. However, the administration of atractyloside (Atr) during IPC nullified this protective effect, highlighting the complex interactions between these treatments. After I/R damage, IPC successfully reduced the decline in endogenous antioxidants (GSH, SOD, and catalase) in normal hearts, but not in HL hearts. AL treatment restored these antioxidant levels in HL hearts, indicating AL may boost antioxidant defences in hyperlipidaemic conditions. However, Atr administration during IPC negated this beneficial effect, suggesting that AL and IPC pathways are susceptible to disruption by Atr.

The results highpoint the complex character of alantolactone (AL) cutting-edge the modulation of mitochondrial function during ischemic preconditioning (IPC) in hyperlipidaemic (HL) rat hearts. In comparison to sham controls, HL rat hearts showed markedly impaired mitochondrial function as measured by formazan levels and TMRM fluorescence intensity.

This impairment indicates that hyperlipidemia exerts a detrimental effect on mitochondrial integrity and activity.

Histopathological analysis showed that IPC reduced inflammation and myonecrosis in normal hearts, with these benefits further enhanced by AL treatment in HL hearts. However, Atr administration during IPC nullified these histological improvements induced by AL, indicating that IPC and AL's protective effects in HL conditions depend on the absence of Atr.

The study found that AL facilitated IPC-induced attenuation of PI3K expression in HL hearts. Post hoc results showed a significant increase in PI3K expression in AL-treated hearts with relation to the sham control. IPC reduced PI3K expression in AL-treated hearts, and AL further enhanced this reduction in HL hearts. The combination of Atr with AL significantly affected IPC-mediated PI3K expression changes, indicating PI3K signalling’s critical role in the observed cardioprotective effects.

CONCLUSION:

The findings imply that Alantolactone (AL) has the potential to enhance the cardioprotective effects of IPC in HL rat hearts, primarily by modulating oxidative stress markers and PI3K expression. However, the presence of atractyloside (Atr) can disrupt these benefits, highlighting the complex interplay between these compounds and the underlying molecular pathways. These results emphasize the importance of understanding the molecular mechanisms involved in cardioprotection to develop effective therapeutic strategies for ischemic heart disease in hyperlipidaemic conditions.

Acknowledgements

The authors would like to thank Kamla Nehru Institute of Management and Technology, Sultanpur for making research facilities available to the writers.

Contribution of the Authors

Conceptualization: AKT, MP. Methodology AKT, MP. Software: RKS, AKJ, CSS. Validation: AKT, RKS, AT. Formal analysis: PSG, MP, CSK. Investigation: AKT. Resources: SKS, AKJ. Data Curation: AKT, MP. Writing - Original Draft: AKT. Writing - Review & Editing: AKT, MP, RKS, SKS, AKJ, CSS.

Conflict of interest

The authors declare that they have no conflicts of interest

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors

Ethical statement

This research was reviewed and approved by the Institutional Animal Ethical Committee with registration number. KNIMT/PHAR/IAEC/18/0

REFERENCES:

[1] Bottardi A, Prado GFA, Lunardi M, Fezzi S, Pesarini G, Tavella D, et al. Clinical Updates in Coronary Artery Disease: A Comprehensive Review. J Clin Med 2024;13. https://doi.org/10.3390/jcm13164600.

[2] Zhao X, Wang D, Qin L. Lipid profile and prognosis in patients with coronary heart disease: a meta-analysis of prospective cohort studies. BMC Cardiovasc Disord 2021;21. https://doi.org/10.1186/s12872-020-01835-0.

[3] Bertero E, Popoiu TA, Maack C. Mitochondrial calcium in cardiac ischemia/reperfusion injury and cardioprotection. Basic Res Cardiol 2024;119:569–85. https://doi.org/10.1007/s00395-024-01060-2.

[4] Mariana M, Cairrao E. Phthalates implications in the cardiovascular system. J Cardiovasc Dev Dis 2020;7. https://doi.org/10.3390/JCDD7030026.

[5] Bennett MR. Basic science APOPTOSIS IN THE CARDIOVASCULAR SYSTEM. n.d.

[6] Hausenloy DJ, Yellon DM. Myocardial ischemia-reperfusion injury: A neglected therapeutic target. Journal of Clinical Investigation 2013;123:92–100. https://doi.org/10.1172/JCI62874.

[7] Zhang H, Hu H, Zhai C, Jing L, Tian H. Cardioprotective Strategies After Ischemia–Reperfusion Injury. American Journal of Cardiovascular Drugs 2024;24:5–18. https://doi.org/10.1007/s40256-023-00614-4.

[8] Hao Y, Xin M, Feng L, Wang X, Wang X, Ma D, et al. Review Cerebral Ischemic Tolerance and Preconditioning: Methods, Mechanisms, Clinical Applications, and Challenges. Front Neurol 2020;11. https://doi.org/10.3389/fneur.2020.00812.

[9] Turer AT, Hill JA. Pathogenesis of myocardial ischemia-reperfusion injury and rationale for therapy. American Journal of Cardiology 2010;106:360–8. https://doi.org/10.1016/j.amjcard.2010.03.032.

[10] Yang X, Cohen M V., Downey JM. Mechanism of cardioprotection by early ischemic preconditioning. Cardiovasc Drugs Ther 2010;24:225–34. https://doi.org/10.1007/s10557-010-6236-x.

[11] Zhang M, Liu Q, Meng H, Duan H, Liu X, Wu J, et al. Ischemia-reperfusion injury: molecular mechanisms and therapeutic targets. Signal Transduct Target Ther 2024;9. https://doi.org/10.1038/s41392-023-01688-x.

[12] Kumar Tiwari A, Gupta PS, Prasad M, of Authors OI, Arun Kumar Tiwari co-A, Author C. Involvement Of Inula Racemose Hook Extract on Ischemic Preconditioning Mediated Cardioprotection in Diabetes-188X. vol. 10. 2023.

[13] Cai Y, Gao K, Peng B, Xu Z, Peng J, Li J, et al. Alantolactone: A Natural Plant Extract as a Potential Therapeutic Agent for Cancer. Front Pharmacol 2021;12. https://doi.org/10.3389/fphar.2021.781033.

[14] Tiwari AK, Gupta PS, Prasad M, Malairajan P. Modulation of Inula racemosa Hook Extract on Cardioprotection by Ischemic Preconditioning in Hyperlipidaemic Rats. J Pharmacopuncture 2022;25:369–81. https://doi.org/10.3831/KPI.2022.25.4.369.

[15] Cai Y, Gao K, Peng B, Xu Z, Peng J, Li J, et al. Alantolactone: A Natural Plant Extract as a Potential Therapeutic Agent for Cancer. Front Pharmacol 2021;12. https://doi.org/10.3389/fphar.2021.781033.

[16] Liu M, Liu P, Zheng B, Liu Y, Li L, Han X, et al. Cardioprotective effects of alantolactone on isoproterenol-induced cardiac injury and cobalt chloride-induced cardiomyocyte injury. Int J Immunopathol Pharmacol 2022;36. https://doi.org/10.1177/20587384211051993.

[17] Ruiz-Meana M. Ischaemic preconditioning and mitochondrial permeability transition: A long-lasting relationship. Cardiovasc Res 2012;96:157–9. https://doi.org/10.1093/cvr/cvs177.

[18] Bouchareb R, CÔté N, Marie-Chloé-Boulanger, Le Quang K, El Husseini D, Asselin J, et al. Carbonic anhydrase XII in valve interstitial cells promotes the regression of calcific aortic valve stenosis. J Mol Cell Cardiol 2015;82:104–15. https://doi.org/10.1016/j.yjmcc.2015.03.002.

[19] I K, N M, Y-J Y, J-Y L. Induce Hyperlipidemia in Rats Using High Fat Diet Investigating Blood Lipid and Histopathology. Journal of Hematology and Blood Disorders 2018;4. https://doi.org/10.15744/2455-7641.4.104.

[20] Srivastav RK, Ansari TM, Prasad M, Vishwakarma VK, Upadhyay PK, Ahsan F, et al. An inquest into regulatory mechanism of caveolin by ischemic preconditioning against orchidectomy-challenged rat heart. Mol Cell Biochem 2021;476:2587–601. https://doi.org/10.1007/s11010-021-04109-1.

[21] Srivastav RK, Ansari TM, Prasad M, Vishwakarma VK, Upadhyay PK, Ahsan F, et al. An inquest into regulatory mechanism of caveolin by ischemic preconditioning against orchidectomy-challenged rat heart. Mol Cell Biochem 2021;476:2587–601. https://doi.org/10.1007/s11010-021-04109-1.

[22] Waterborg JH. The Lowry Method 7 7 From: The Protein Protocols Handbook, 2nd Edition The Lowry Method for Protein Quantitation. n.d.

[23] Kamboj SS, Kumar V, Kamboj A, Sandhir R. Mitochondrial oxidative stress and dysfunction in rat brain induced by carbofuran exposure. Cell Mol Neurobiol 2008;28:961–9. https://doi.org/10.1007/s10571-008-9270-5.

[24] Jaiswal A, Kumar S, Enjamoori R, Seth S, Dinda AK, Maulik SK. Peripheral benzodiazepine receptor ligand Ro5-4864 inhibits isoprenaline-induced cardiac hypertrophy in rats. Eur J Pharmacol 2010;644:146–53. https://doi.org/10.1016/j.ejphar.2010.06.058

Therapeutic Modulation of Ischemic Preconditioning by Alantolactone in Diseased Rat Hearts

Article Information:

Abstract:

Keywords:

Article :

INTRODUCTION:

II. MATERIAL AND METHOD:

RESULTS:

DISCUSSION:

CONCLUSION:

REFERENCES:

Article Citation

APA

MLA

Chicago

Harvard

Vancouver