Comparative Study on the Role of Berberine and Berberis lycium Royle Roots Extract against the Biochemical Markers and Cyclin D1 Expression in HCC Animal Model

Diethylamine nitrosamine (DEN), as an initiator of liver tumor, and carbon tetrachloride (CCl4), as a tumor promoter, have been used to study the molecular events of liver cancer in animal models. Recently, our in vitro study reported BLE (Berberis lycium Royle ethanol extract) as the most effective agent against liver cancer, thus we continued our study in vivo to assess the hepatoprotective effect of BLE and its most active alkaloid, berberine, in albino mice (70 male). Moreover, we investigated the biochemical/immunohistochemical effects of a single alkaloid versus the effect of Berberis extract in mice liver. Hepatic cancer was induced in mice by a single intraperitoneal injection with DEN (100 mg/kg b.wt), followed by biweekly injections of CCl4 (0.5 mL/kg) for 30 days. The development of liver cancer was assessed after 60 days of DEN injection by measuring the elevated level of the serum tumor marker alpha-fetoprotein (AFP) and liver function test (ALT, AST, ALP, and BUN) markers. After the confirmation of liver cancer development, the BLE extract and berberine were fed to mice for 90 days and the serum biomarkers for liver injury (LFTs and AFP) were measured again. Overall, berberine (120 mg/kg b.wt) proved to be a stronger agent in reducing the symptoms of HCC in mice, as compared with BLE. Histopathological analysis agreed well with the biochemical observations. Immunohistochemistry analysis suggested significant suppression of the quantitative expression of the key oncogene cyclin D1 at low (60 mg/kg) and high (120 mg/kg) doses of berberine. These findings implicate the amelioration of hepatocarcinoma by berberine more prominently in mice, by suppression of cyclin-dependent kinase activator (CD1) expression, reducing LFTs, as well as AFP, in the serum. Thus, our findings are novel, as berberine may help in controlling the perturbation in CD1 associated with aggressive forms of HCC. However, future studies should be directed at finding out whether berberine has any effect on inhibitors (p27 and CDKI) of cyclin-dependent kinase too.

Ethnobotanical and Biochemical Study of Berberis lycium Royle Collected from Different Areas of Azad Jammu and Kashmir

Berberis lycium Royle has a long history of medicinal uses to treat different diseases. It naturally grows on the mountains of Indian subcontinent. Its ethnobotanical and biochemical study from the state of Azad Jammu and Kashmir (AJ&K) was not previously explored. So, the objective of the current study was to explore the ethnobotanical and biochemical properties of the B. lycium Royle population of AJ&K. For this purpose, samples of B. lycium Royle were randomly collected from five districts of Azad Jammu and Kashmir, including thirty-five locations. Demographic features of informants such as plant part used, methods of preparation, modes of administration, conservation status, and ethnomedicinal uses were documented. It was used for treating different diseases such as diabetes, arthritis, joint pain, and stomach ulcer. This plant is very famous for providing medicinal roots, leaves, and fruits which are extensively used in many parts of the world. The biochemical analysis was conducted for total phenolic contents (TPC), chlorophyll contents, and antioxidant activity. The highest level of TPC found was 88.66 ± 1.07 µg/g of gallic acid equivalent phenolic (GAE) from leaves collected from Patikka (Chanjhal), Muzaffarabad District, AJ&K. The highest total chlorophyll contents (3.75 ± 0.53 µg/ml) were found in samples collected from Sathrian, Neelum District. The highest antioxidant activity with lowest IC50 value (33.26 µg/ml) was obtained from the root of sample collected from Bakreyali, Muzaffarabad District, as compared with other districts. The concentration of berberine was found to be 4.76 percent in the root bark of B. lycium Royle, estimated by high-performance liquid chromatography (HPLC). In syrup composition, 0.95 mg/5 ml of berberine was used. Hence, it is concluded that amongst the five districts, the plant parts (stem, fruits, and root) collected from Muzaffarabad District, AJ&K, showed the highest medicinal potential due to its unique climatic conditions.

Reproductive Biology and Pollination Ecology of Berberis lycium Royle: A Highly Valued Shrub of Immense Medicinal Significance

Study of reproductive biology and pollination ecology helps in understanding the life history patterns of species. Such a study brings to light the bottlenecks, if any, on account of which the individuals of the species are not able to reproduce in nature and ultimately helps in planning appropriate conservation strategies for the species under threat. The present study was aimed at examining the morphological and reproductive variance in Berberis lycium, a threatened ecological specialist growing within shrubberies and open hillsides of the North-Western Himalayas in India. B. lycium displays three different variants. Flowering period ranges from February to September. Pollen viability as reported on fluorescein diacetate and acetocarmine treatments was highest for variant I, while maximum pollen output was obtained for variant III. Pollen pistil interaction is brought by the movement of anther towards stigma. Fluorescence microscopy of hand pollinated club shaped stigma shows that the germinating pollen form a ring over the receptive adaxial surface. Pollination syndrome is entomophily. Variant II attracts a significantly large number of pollinators from diverse insect families. Breeding experiments reflect that plants are self-compatible and cross fertile. Reproductive output (% fruit set) was highest for variant II followed by III and I, respectively. This investigation helped to understand the effect of different biotic and abiotic constraints on the phenology and reproductive biology of the plant. The information generated so will enable conservationists to design appropriate strategies for its long-term survival and sustenance in nature.

Antiulcerogenic effect of Methanolic Root Extract of Berberis lycium Linn by Pylorus ligation and Ethanol induced Ulceration

Background: Gastric ulcer is one of the most prevalent gastrointestinal disorders, which affects approximately 5-10% of people during their life. In recent years, abundant work has been carried out on herbal medicine to clarify their potential efficacy in gastric ulcer prevention or management. The present study was carried out to evaluate the antiulcer activity of the methanolic root extract of Berberis lycium in albino rats. Method: The methanolic root extract of Berberis lycium was prepared by hot extraction method. Anti-ulcer activity was evaluated and method employed was pylorus ligation and ethanol induced in albino rats. Preliminary methanolic extract of Berberis lycium was subjected to the acute oral toxicity study according to the OECD guideline no. 425. Animals were divided into four groups of six animals each. The animals of Group I served as normal control (vehicle) which received distilled water. Group II and III received 250 mg/kg and 500 mg/kg of methanolic root extract, respectively. In pylorus ligation induced ulcer model, various parameters were studied viz. gastric volume, pH, total acidity, free acidity, and ulcer index. Ulcer index and percentage inhibition of ulceration was determined for ethanol induced ulcer model. Group IV received Ranitidine at 50 mg/kg was used as the standard drug. Pretreatment of methanol root extract of Berberis lycium showed significant (P˂0.05) decrease in the gastric volume, total acidity and free acidity. However, pH of the gastric juice was significantly increased only at higher dose 500 mg/kg. It showed also significant (P˂0.05) decrease in number of ulcers and ulcer score index in pylorus ligation and ethanol induced ulceration models. Results: The methanol root extract of Berberis lycium showed a significant reduction in the total acidity, free acidity and acid volume. The efficacy of plant extract at high dose was comparable with the standard drug Ranitidine. Conclusion: Our study results support the ethnomedical use of root of Berberis lycium. Keywords: Antiulcer activity, Berberis lycium, Pylorus ligation, Ranitidine, Ulcer Index.

Reduction of liver enzyme ALT by berberis lycium in acetaminophen induced hepatic damage in mice liver.

Objective: The objective of this research was to study the effect of Berberis lyceum (herbal plant) in reduction of liver enzyme ALT in mice. Study Design: Quasi- experimental Study. Setting: Peshawar Medical College, Peshawar. Period: December, 2016 to May, 2017. Materials & Methods: It was an experimental study which was carried out at Peshawar Medical College Warsak Road Peshawar. Total 30 (thirty) mice were used divided in six different groups with five mice in each group. After inducing hepatotoxicity with acetaminophen in the mice, blood samples were taken and LFTs were performed to observe serum ALT values and the effects of different doses of the plant extract were evaluated. Results: The ALT levels of negative control and experimental group were compared and it was shown that ALT levels in experimental groups were 232±42.3, 143±32.5 and 62.2±18.2 whereas serum ALT value of negative control group was 571.4±123.3 which showed that ALT had been decreased by increasing the dose of plant extract. Conclusion: It was concluded that Berberis Lycium, a herbal plant has a potentially active hepato protective role in bringing serum ALT to normal. This plant can be useful in reversing the hepatotoxicity in drug induced elevation of liver enzyme ALT.